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1.  Involvement of DNA polymerase beta overexpression in the malignant transformation induced by benzo[a]pyrene 
Toxicology  2013;309:73-80.
Objective
To explore the relationship between DNA polymerase β (pol β) overexpression and benzo[a]pyrene (BaP) carcinogenesis.
Methods
Firstly, mouse embryonic fibroblasts that express wild-type level of DNA polymerase β (pol β cell) and high level of pol β (pol β oe cell) were treated by various concentrations of BaP to determine genetic instability induced by BaP under differential expression levels of pol β. Secondly, malignant transformation of pol β cells by low concentration of BaP (20 μM) was determined by soft agar colony formation assay and transformation focus assay. Thirdly, the mRNA and protein levels of BaP-transformed pol β cells (named pol β-T cells) was measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot, and the genetic instability of these cells were examined by HPRT gene mutation assay and random amplified polymorphic DNA (RAPD) assay.
Results
Pol β cells were successfully transformed into malignant pol β-T cells by an exposure to low concentration of BaP for 6 months. Pol β-T cells exhibited increased levels of pol β gene expression, HPRT gene mutation frequency and polymorphisms of RAPD products that were comparable to those of pol β oe cells.
Conclusion
Pol β overexpression and its-associated genetic instability may play a key role in BaP carcinogenesis.
doi:10.1016/j.tox.2013.04.017
PMCID: PMC4281483  PMID: 23652152
DNA polymerase β; benzo[a]pyrene; malignant transformation; base excision repair; genetic instability; carcinogenesis
2.  Genetic Diversity of Bacterial Communities and Gene Transfer Agents in Northern South China Sea 
PLoS ONE  2014;9(11):e111892.
Pyrosequencing of the 16S ribosomal RNA gene (rDNA) amplicons was performed to investigate the unique distribution of bacterial communities in northern South China Sea (nSCS) and evaluate community structure and spatial differences of bacterial diversity. Cyanobacteria, Proteobacteria, Actinobacteria, and Bacteroidetes constitute the majority of bacteria. The taxonomic description of bacterial communities revealed that more Chroococcales, SAR11 clade, Acidimicrobiales, Rhodobacterales, and Flavobacteriales are present in the nSCS waters than other bacterial groups. Rhodobacterales were less abundant in tropical water (nSCS) than in temperate and cold waters. Furthermore, the diversity of Rhodobacterales based on the gene transfer agent (GTA) major capsid gene (g5) was investigated. Four g5 gene clone libraries were constructed from samples representing different regions and yielded diverse sequences. Fourteen g5 clusters could be identified among 197 nSCS clones. These clusters were also related to known g5 sequences derived from genome-sequenced Rhodobacterales. The composition of g5 sequences in surface water varied with the g5 sequences in the sampling sites; this result indicated that the Rhodobacterales population could be highly diverse in nSCS. Phylogenetic tree analysis result indicated distinguishable diversity patterns among tropical (nSCS), temperate, and cold waters, thereby supporting the niche adaptation of specific Rhodobacterales members in unique environments.
doi:10.1371/journal.pone.0111892
PMCID: PMC4218858  PMID: 25364820
3.  Diagnosis of infantile myofibromatosis with pseudo-ulcerated plaque using prenatal ultrasound: A case report 
The present case report describes a case of infantile myofibromatosis (IM) with a pseudo-ulcerated plaque on the right side of the back of a fetus, detected in the 38th week of gestation using prenatal ultrasound. The fetus was examined weekly by ultrasound to measure the size of the mass. At birth, the scarlet mass was slightly elevated compared with the skin around it, with a cavity in the center. It appeared similar to an ulcerated plaque, but the surface of the mass was intact and smooth with a stratum lucidum. Thus, the mass was indicated to be a pseudo-ulcerated plaque. Three months later, the mass had grown larger and so was removed by surgery. The pathology of the mass was confirmed as IM. It is suggested that IM should be considered when a soft tissue tumor is presented by prenatal ultrasound.
doi:10.3892/etm.2014.2010
PMCID: PMC4218696  PMID: 25371730
infantile myofibromatosis; pseudo-ulcerated plaque; ultrasound; magnetic resonance imaging; prenatal
4.  Atorvastatin prevents RhoC isoprenylation, invasion, and metastasis in human melanoma cells 
Molecular cancer therapeutics  2003;2(10):941-948.
Melanoma is a deadly cancer due to its propensity to metastasize. Pharmacological inhibition of cell motility may benefit patients with cutaneous melanoma by preventing metastasis to internal organs. The Rho GTPases are signaling molecules that drive metastasis by controlling cell motility. We found RhoC to be expressed in clinical melanoma specimens and hypothesized that inhibiting its activation might prevent metastasis. Some Rho proteins, such as RhoC, depend on posttranslational geranylgeranylation for biological activity. We investigated the effect that Atorvastatin, a 3-hydroxy 3-methylglutaryl CoA (HMG-CoA) reductase inhibitor that prevents Rho geranylgeranylation, had on subcellular localization and activity of Rho proteins as well as the metastatic ability of melanoma cells. Atorvastatin inhibited Rho activation and reverted the metastatic phenotype of human melanoma cells in vitro. Moreover, Atorvastatin, at plasma levels comparable to those used to treat of hypercholesterolemia, inhibited in vivo metastasis of melanoma cells overexpressing RhoC. These results support further examination of statins for primary prophylaxis of melanoma metastasis.
PMCID: PMC4161126  PMID: 14578459
5.  Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene 
PLoS ONE  2014;9(9):e106864.
Background
Pigs are an optimal animal for conducting biomedical research because of their anatomical and physiological resemblance to humans. In contrast to the abundant resources available in the study of mice, few fluorescent protein-harboring porcine models are available for preclinical studies. In this paper, we report the successful generation and characterization of a transgenic DsRed-Monomer porcine model.
Methods
The transgene comprised a CMV enhancer/chicken-beta actin promoter and DsRed monomeric cDNA. Transgenic pigs were produced by using pronuclear microinjection. PCR and Southern blot analyses were applied for identification of the transgene. Histology, blood examinations and computed tomography were performed to study the health conditions. The pig amniotic fluid progenitor/stem cells were also isolated to examine the existence of red fluorescence and differentiation ability.
Results
Transgenic pigs were successfully generated and transmitted to offspring at a germ-line transmission rate of 43.59% (17/39). Ubiquitous expression of red fluorescence was detected in the brain, eye, tongue, heart, lung, liver, pancreas, spleen, stomach, small intestine, large intestine, kidney, testis, and muscle; this was confirmed by histology and western blot analyses. In addition, we confirmed the differentiation potential of amniotic fluid progenitor stem cells isolated from the transgenic pig.
Conclusions
This red fluorescent pig can serve as a host for other fluorescent-labeled cells in order to study cell-microenvironment interactions, and can provide optimal red-fluorescent-labeled cells and tissues for research in developmental biology, regenerative medicine, and xenotransplantation.
doi:10.1371/journal.pone.0106864
PMCID: PMC4154781  PMID: 25187950
6.  Bacteria repelling poly(methylmethacrylate-co-dimethylacrylamide) coatings for biomedical devices† †Electronic supplementary information (ESI) available: Polymer microarray screening, including analysis of bacterial adhesion by fluorescence microscopy and SEM, and chemical composition of bacteria repelling polymers identified in the screen; polymer synthesis and characterisation; preparation of catheter pieces and solvent studies, and details for confocal imaging/analysis. See DOI: 10.1039/c4tb01129e Click here for additional data file.  
A polymer microarray screen identified poly(methylmethacrylate-co-dimethylacrylamide) as a bacteria repelling surface, which substantially reduced binding of bacteria on coated catheters.
Nosocomial infections due to bacteria have serious implications on the health and recovery of patients in a variety of medical scenarios. Since bacterial contamination on medical devices contributes to the majority of nosocomical infections, there is a need for redesigning the surfaces of medical devices, such as catheters and tracheal tubes, to resist the binding of bacteria. In this work, polyurethanes and polyacrylates/acrylamides, which resist binding by the major bacterial pathogens underpinning implant-associated infections, were identified using high-throughput polymer microarrays. Subsequently, two ‘hit’ polymers, PA13 (poly(methylmethacrylate-co-dimethylacrylamide)) and PA515 (poly(methoxyethylmethacrylate-co-diethylaminoethylacrylate-co-methylmethacrylate)), were used to coat catheters and substantially shown to decrease binding of a variety of bacteria (including isolates from infected endotracheal tubes and heart valves from intensive care unit patients). Catheters coated with polymer PA13 showed up to 96% reduction in bacteria binding in comparison to uncoated catheters.
doi:10.1039/c4tb01129e
PMCID: PMC4247239  PMID: 25580245
7.  The association of metabolic syndrome and its factors with gallstone disease 
BMC Family Practice  2014;15:138.
Background
To investigate the association between metabolic syndrome, including its factors, and gallstone disease (GSD) in a Taiwanese population.
Methods
We conducted a cross-sectional study during 2011 ~ 2012. A total of 12050 subjects who completed a questionnaire and underwent physical examination, laboratory tests and abdominal ultrasonography formed the study population.
Results
The prevalences of metabolic syndrome and gallstone disease were 24.09% and 6.16%. In an age- and sex-adjusted logistic regression model, metabolic syndrome was associated with gallstone disease (OR = 1.61; P < 0.0001). Age, abdominal obesity, and lower high-density lipoprotein cholesterol were associated with gallstone disease after adjusting for other factors. Females had a higher odds ratio than males in waist circumference for GSD, whereas males had a lower odds ratio than females in HDL-C for GSD.
Conclusions
The present study suggests that metabolic syndrome is related to gallstone disease. Waist circumference and high-density lipoprotein cholesterol are all associated with GSD. Men and women may possibly have different priorities and strategies to reduce the burden of GSD.
doi:10.1186/1471-2296-15-138
PMCID: PMC4118643  PMID: 25070766
Obesity; Dyslipidemia; Metabolic factors; Metabolic syndrome; Gallstone disease
8.  Rechallenge with pemetrexed-based chemotherapy improves the survival of patients with advanced non-squamous non-small-cell lung cancer 
Molecular and Clinical Oncology  2014;2(6):953-959.
Rechallenge chemotherapy with pemetrexed was shown to be efficient in malignant pleural mesothelioma; however, its role in non-small-cell lung cancer (NSCLC) has not been investigated. In this study, we retrospectively enrolled 31 patients with non-squamous NSCLC who had achieved disease control with initial pemetrexed treatment, followed by rechallenge with pemetrexed-based chemotherapy (PBC) upon disease progression. After the rechallenge, 5 patients (16.1%) achieved partial remission (PR), 17 (54.8%) achieved stable disease (SD) and 9 (29.1%) experienced progressive disease. The treatment was generally well tolerated, with a low rate of toxicity. The median progression-free survival (PFS) was 3.8 months with the rechallenge. Patients with a PFS of ≥10 months with initial PBC exhibited longer PFS and overall survival (OS) with the rechallenge compared to those with a PFS of <10 months with initial PBC (PFS: 6.2±0.33 vs. 3.1±0.26 months, respectively; P=0.011; and OS, 19.8±3.2 vs. 9.2±1.1 months, respectively; P=0.005). The time from the termination of initial PBC to disease progression was also associated with survival after the rechallenge. However, the response to initial PBC (PR vs. SD) did not affect PFS after the rechallenge. No significant differences were observed in thymidylate synthase expression, echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase gene fusion, or epithelial growth factor receptor mutation status between pemetrexed-sensitive and pemetrexed-resistant patients. Our results demonstrated that rechallenge with PBC was well tolerated and survival after the rechallenge was associated with survival during initial PBC. Therefore, patients with a PFS of ≥10 months or time-to-disease progression ≥3 months may be considered as candidates for pemetrexed rechallenge.
doi:10.3892/mco.2014.359
PMCID: PMC4179803  PMID: 25279180
pemetrexed; rechallenge; non-small-cell lung cancer; overall survival; adverse effects
9.  Autoantibody signatures involving glycolysis and splicesome proteins precede a diagnosis of breast cancer among post-menopausal women 
Cancer research  2012;73(5):1502-1513.
We assessed the autoantibody repertoire of a mouse model engineered to develop breast cancer and the repertoire of autoantibodies in plasmas collected at a pre-clinical time point and at the time of clinical diagnosis of breast cancer. In seeking to identify common pathways, networks and protein families associated with the humoral response, we elucidated the dynamic nature of tumor antigens and autoantibody interactions. Lysate proteins from an immortalized cell line from an MMTV-neu mouse model and from MCF7 human breast cancers were spotted onto nitrocellulose microarrays and hybridized with mouse and human plasma samples, respectively. Ig-based plasma immunoreactivity against glycolysis and spliceosome proteins was a predominant feature observed both in tumor bearing mice and in pre-diagnostic human samples. Interestingly, autoantibody reactivity was more pronounced further away than closer to diagnosis. We provide evidence for dynamic changes in autoantibody reactivity with tumor development and progression that may depend in part on the extent of antigen-antibody interactions.
doi:10.1158/0008-5472.CAN-12-2560
PMCID: PMC4085738  PMID: 23269276
Breast cancer; Autoantibody; Glycolysis; Spliceosome; Pre-diagnostic
10.  Body Burden of Hg in Different Bio-Samples of Mothers in Shenyang City, China 
PLoS ONE  2014;9(5):e98121.
Hg is an accumulative and neuro-toxic heavy metal which has a wide range of adverse effects in human health. However, few studies are available on body burden of Hg level in different bio-samples of pregnant women in Chinese population. Therefore, this study evaluated Hg levels in different maternal bio-samples in Shenyang city, China and investigated the correlation of Hg levels in different bio-samples. From October to December 2008, 200 pregnant women about to deliver their babies at ShengJing Hospital (Shenyang city, northeast of China) participated in this study. The geometric mean (GM) of Hg levels in cord blood, maternal venous blood, breast milk, and maternal urine were 2.18 µg/L, 1.17 µg/L, 1.14 µg/L, and 0.73 µg/L, respectively, and the GM of maternal hair Hg level was 404.45 µg/kg. There was a strong correlation between cord blood and maternal blood total Hg level (r = 0.713, P<0.001). Frequency of fish consumption more than or equal to 3 times per week during pregnancy was suggested as a significant risk factor of prenatal Hg exposure (unadjusted OR 3.5, adjusted OR 2.94, P<0.05). This study provides evidence about Hg burden of mothers and the risk factors of prenatal Hg exposure in Shenyang city, China.
doi:10.1371/journal.pone.0098121
PMCID: PMC4032259  PMID: 24858815
11.  A Ten-MicroRNA Signature Identified from a Genome-Wide MicroRNA Expression Profiling in Human Epithelial Ovarian Cancer 
PLoS ONE  2014;9(5):e96472.
Epithelial ovarian cancer (EOC) is the most common gynecologic malignancy. To identify the micro-ribonucleic acids (miRNAs) expression profile in EOC tissues that may serve as a novel diagnostic biomarker for EOC detection, the expression of 1722 miRNAs from 15 normal ovarian tissue samples and 48 ovarian cancer samples was profiled by using a quantitative real-time polymerase chain reaction (qRT-PCR) assay. A ten-microRNA signature (hsa-miR-1271-5p, hsa-miR-574-3p, hsa-miR-182-5p, hsa-miR-183-5p, hsa-miR-96-5p, hsa-miR-15b-5p, hsa-miR-182-3p, hsa-miR-141-5p, hsa-miR-130b-5p, and hsa-miR-135b-3p) was identified to be able to distinguish human ovarian cancer tissues from normal tissues with 97% sensitivity and 92% specificity. Two miRNA clusters of miR183-96-183 (miR-96-5p, and miR-182, miR183) and miR200 (miR-141-5p, miR200a, b, c and miR429) are significantly up-regulated in ovarian cancer tissue samples compared to those of normal tissue samples, suggesting theses miRNAs may be involved in ovarian cancer development.
doi:10.1371/journal.pone.0096472
PMCID: PMC4015980  PMID: 24816756
12.  Computational Analysis of mRNA Expression Profiles Identifies the ITG Family and PIK3R3 as Crucial Genes for Regulating Triple Negative Breast Cancer Cell Migration 
BioMed Research International  2014;2014:536591.
Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer that does not express estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (Her2/neu). TNBC has worse clinical outcomes than other breast cancer subtypes. However, the key molecules and mechanisms of TNBC migration remain unclear. In this study, we compared two normalized microarray datasets from GEO database between Asian (GSE33926) and non-Asian populations (GSE46581) to determine the molecules and common pathways in TNBC migration. We demonstrated that 16 genes in non-Asian samples and 9 genes in Asian samples are related to TNBC migration. In addition, our analytic results showed that 4 genes, PIK3R3, ITGB1, ITGAL, and ITGA6, were involved in the regulation of actin cytoskeleton. Our results indicated potential genes that link to TNBC migration. This study may help identify novel therapeutic targets for drug development in cancer therapy.
doi:10.1155/2014/536591
PMCID: PMC4032728  PMID: 24982892
13.  Distributed SLAM Using Improved Particle Filter for Mobile Robot Localization 
The Scientific World Journal  2014;2014:239531.
The distributed SLAM system has a similar estimation performance and requires only one-fifth of the computation time compared with centralized particle filter. However, particle impoverishment is inevitably because of the random particles prediction and resampling applied in generic particle filter, especially in SLAM problem that involves a large number of dimensions. In this paper, particle filter use in distributed SLAM was improved in two aspects. First, we improved the important function of the local filters in particle filter. The adaptive values were used to replace a set of constants in the computational process of importance function, which improved the robustness of the particle filter. Second, an information fusion method was proposed by mixing the innovation method and the number of effective particles method, which combined the advantages of these two methods. And this paper extends the previously known convergence results for particle filter to prove that improved particle filter converges to the optimal filter in mean square as the number of particles goes to infinity. The experiment results show that the proposed algorithm improved the virtue of the DPF-SLAM system in isolate faults and enabled the system to have a better tolerance and robustness.
doi:10.1155/2014/239531
PMCID: PMC4030567  PMID: 24883362
14.  A Polymorphism of ORAI1 rs7135617, Is Associated with Susceptibility to Rheumatoid Arthritis 
Mediators of Inflammation  2014;2014:834831.
Rheumatoid arthritis (RA), a chronic inflammatory disease usually occurring in synovial tissues and joints, is highly associated with genetic and environmental factors. ORAI1, a gene related to cellular immune system, has been shown to be involved in the pathogenesis of chronic inflammatory diseases and immune diseases. To identify whether ORAI1 gene contributes to RA susceptibility, we enrolled 400 patients with RA and 621 healthy individuals for a case-control genetic association study. Five tagging single nucleotides polymorphisms (tSPNs) within ORAI1 gene were selected for genotyping. An SNP, rs7135617, showed a significant correlation with the risk of RA. Our results indicated that genetic polymorphism of ORAI1 gene is involved in the susceptibility of RA in a Taiwanese population.
doi:10.1155/2014/834831
PMCID: PMC3997980  PMID: 24808640
15.  Anthelmintic Activities of Aporphine from Nelumbo nucifera Gaertn. cv. Rosa-plena against Hymenolepis nana 
Nelumbo nucifera Gaertn. cv. Rosa-plena (Nelumbonaceae), commonly known as lotus, is a perennial aquatic plant grown and consumed throughout Asia. All parts of N. nucifera have been used for various medicinal purposes in oriental medicine. From the leaves of Nelumbo nucifera Gaertn. cv. Rosa-plena (an aquatic plant), liriodenine (1), lysicamine (2), (−)-anonaine (3), (-)-asimilobine (4), (-)-caaverine (5), (-)-N-methylasimilobine (6), (-)-nuciferine (7), (-)-nornuciferine (8), (-)-roemerine (9), 7-hydroxydehydronuciferine (10) and cepharadione B (11) were isolated and identification and anthelmintic activities of aporphine was evaluated against Anisakis simplex and Hymenolepis nana. This study found that the above constituents killed H. nana or reduced their spontaneous movements (oscillation/peristalsis). However, the above constituents at various concentrations demonstrated no larvicidal effect or ability to halt spontaneous parasite movement for 72 h against A. simplex, respectively. In addition, according to an assay of cestocidal activity against H. nana and nematocidal activity against A. simplex, we found that the above compounds showed greater lethal efficacy on H. nana than against A. simplex. Further investigation showed that these above constituents have effects against peroxyl radicals under cestocidal effect. Together, these findings suggest that these constituents of Nelumbo nucifera Gaertn. cv. Rosa-plena might be used as anthelmintic agents against H. nana.
doi:10.3390/ijms15033624
PMCID: PMC3975358  PMID: 24583851
Nelumbo nucifera Gaertn; aporphine; Hymenolepis nana; Anisakis simplex; anthelmintic activity; peroxyl radical
16.  Elevation of serum CXCL16 level correlates well with atherosclerotic ischemic stroke 
Introduction
Currently there are no reliable biological markers for ischemic stroke. The novel chemokine CXCL16 is known to be involved in the development of atherosclerosis. Nevertheless, the real role of CXCL16 in atherosclerotic disorders remains uncertain. The goal of our study was to investigate the associations between serum-soluble CXCL16 level and atherosclerotic ischemic stroke, including large artery atherosclerosis (LAA) and small artery occlusion (SAO) subtypes, and to explore whether elevation in CXCL16 levels is correlated with the severity of large arterial stenosis.
Material and methods
The study recruited 227 subjects, including 74 controls and 153 consecutive patients with acute ischemic stroke from atherosclerosis of the carotid artery. The etiology of the acute ischemic strokes was classified into LAA (n = 86) subtype and SAO (n = 67) subtype according to the TOAST criteria, and the severity of carotid artery stenosis was assessed by the NASCET criteria. Serum-soluble CXCL16 concentration was measured by enzyme-linked immunosorbent assay.
Results
Serum CXCL16 concentrations were significantly increased in both LAA (2.36 ng/ml) and SAO subtypes (2.13 ng/ml) when compared to that of the controls (2.04 ng/ml, p < 0.01 and p < 0.05, respectively), and it was significantly elevated in LAA subtype than in SAO subtype (p < 0.05). However, significant differences in CXCL16 levels between the high-grade stenosis group (2.36 ng/ml) and moderate-grade stenosis group (2.24 ng/ml) of LAA subtype were not found (p > 0.05). A correlation of serum levels of CXCL16 with serum levels of hs-CRP, fibrinogen and lipid parameters was not observed (p > 0.05).
Conclusions
Increased serum level of soluble CXCL16 was independently associated with atherosclerotic ischemic stroke, particularly LAA subtype.
doi:10.5114/aoms.2013.39200
PMCID: PMC3953970  PMID: 24701213
CXCL16; chemokine; atherosclerosis; ischemic stroke; inflammation
17.  Spatial Repolarization Heterogeneity Detected by Magnetocardiography Correlates with Cardiac Iron Overload and Adverse Cardiac Events in Beta-Thalassemia Major 
PLoS ONE  2014;9(1):e86524.
Background
Patients with transfusion-dependent beta-thalassemia major (TM) are at risk for myocardial iron overload and cardiac complications. Spatial repolarization heterogeneity is known to be elevated in patients with certain cardiac diseases, but little is known in TM patients. The purpose of this study was to evaluate spatial repolarization heterogeneity in patients with TM, and to investigate the relationships between spatial repolarization heterogeneity, cardiac iron load, and adverse cardiac events.
Methods and Results
Fifty patients with TM and 55 control subjects received 64-channel magnetocardiography (MCG) to determine spatial repolarization heterogeneity, which was evaluated by a smoothness index of QTc (SI-QTc), a standard deviation of QTc (SD-QTc), and a QTc dispersion. Left ventricular function and myocardial T2* values were assessed by cardiac magnetic resonance. Patients with TM had significantly greater SI-QTc, SD-QTc, and QTc dispersion compared to the control subjects (all p values<0.001). Spatial repolarization heterogeneity was even more pronounced in patients with significant iron overload (T2*<20 ms, n = 20) compared to those with normal T2* (all p values<0.001). Loge cardiac T2* correlated with SI-QTc (r = −0.609, p<0.001), SD-QTc (r = −0.572, p<0.001), and QTc dispersion (r = −0.622, p<0.001), while all these indices had no relationship with measurements of the left ventricular geometry or function. At the time of study, 10 patients had either heart failure or arrhythmia. All 3 indices of repolarization heterogeneity were related to the presence of adverse cardiac events, with areas under the receiver operating characteristic curves (ranged between 0.79 and 0.86), similar to that of cardiac T2*.
Conclusions
Multichannel MCG demonstrated that patients with TM had increased spatial repolarization heterogeneity, which is related to myocardial iron load and adverse cardiac events.
doi:10.1371/journal.pone.0086524
PMCID: PMC3903540  PMID: 24475137
18.  Expression analysis of BMP2, BMP5, BMP10 in human colon tissues from Hirschsprung disease patients 
Objective: Bone morphogenetic proteins (BMPs) are members of the transforming growth factor β (TGF β) superfamily. BMP2, BMP5 and BMP10 exert their biological functions by interacting with membrane bound receptors belonging to the serine/threonine kinase family. Hirschsprung disease (HSCR) is characterized by the absence of intramural ganglion cells in the nerve plexuses of the distal gut. However, putative Notch function in enteric nervous system (ENS) development and the etiology of HSCR is unknown. Methods: Aganglionic and ganglionic colon segment tissues of 50 HSCR patients were investigated for the expression pattern of BMP2, BMP5 and BMP10 using real-time RT-PCR, Western blot analysis and immunohistochemical staining. Results: The mRNA levels of BMP2, BMP5 and BMP10 in the stenotic colon segment from HSCR patients were significantly higher than those in the normal ones. Similar increased expressions of them in the stenotic colon segments were detected by Western blotting coupled with densitometry analysis. Lastly, immunohistologicl stain showed significant BMP2, 5 and 10 increases in mucous and muscular layers from stenotic colon segments compared to normal segments. Conclusions: BMP2, BMP5 and BMP10 are elevated in the stenotic colon segment of HSCR, and BMPs signaling plays a pivotal role in the development of HSCR.
PMCID: PMC3925897  PMID: 24551273
Bone morphogenetic proteins (BMPs); Hirschsprung disease (HSCR); enteric nervous system (ENS); stenotic colon; ganglion cell
19.  High immunogenicity of nicotine vaccines obtained by intradermal delivery with safe adjuvants 
Vaccine  2012;31(1):159-164.
Immunotherapy for tobacco addiction may offer a safe, alternative treatment if the immunogenicity of the current nicotine vaccines can be improved. We show here that intradermal (ID) immunization induces the production of antibody directed against nicotine (NicAb) at a much higher level than conventional intramuscular (IM) immunization. The magnitude and duration of NicAb production was further increased robustly by non-inflammatory laser vaccine adjuvant (LVA), slightly inflammatory monophosphoryl lipid A (MPL) or a combination of MPL and CpG adjuvants. Consequently, significantly fewer vaccination doses were required to attain a high level of NicAb production for an extended period of time and reduce nicotine entry into the brain in the presence of LVA, MPL or MPL/CpG adjuvant, respectively. Yet, the potency of these adjuvants to augment ID nicotine vaccine immunogenicity came at the expense of local skin reactogenicity, with LVA causing little skin reaction and MPL/CpG stimulating overt skin irritation. These observations underscore a necessity of a balance between optimal adjuvant potency and undesired local reactogenicity. In summary, our study presents a novel approach to significantly improve nicotine vaccine immunogenicity by a combination of safe cutaneous vaccine adjuvants with ID immunization.
doi:10.1016/j.vaccine.2012.10.069
PMCID: PMC3557791  PMID: 23123021
nicotine vaccine; adjuvant; intradermal delivery; immunogenicity; addiction; immunotherapy
20.  Spontaneous Electrical Activity of Cultured Interstitial Cells of Cajal from Mouse Urinary Bladder 
Interstitial cells of Cajal (ICCs) from the urinary bladder regulate detrusor smooth muscle activities. We cultured ICCs from the urinary bladder of mice and performed patch clamp and intracellular Ca2+ ([Ca2+]i) imaging to investigate whether cultured ICCs can be a valuable tool for cellular functional studies. The cultured ICCs displayed two types of spontaneous electrical activities which are similar to those recorded in intact bladder tissues. Spontaneous electrical activities of cultured ICCs were nifedipine-sensitive. Carbachol and ATP, both excitatory neurotransmitters in the urinary bladder, depolarized the membrane and increased the frequency of spike potentials. Carbachol increased [Ca2+]i oscillations and basal Ca2+ levels, which were blocked by atropine. These results suggest that cultured ICCs from the urinary bladder retain rhythmic phenotypes similar to the spontaneous electrical activities recorded from the intact urinary bladder. Therefore, we suggest that cultured ICCs from the urinary bladder may be useful for cellular and molecular studies of ICCs.
doi:10.4196/kjpp.2013.17.6.531
PMCID: PMC3874441  PMID: 24381503
Interstitial cells of Cajal; Spontaneous electrical activities; Urinary bladder
21.  Manual Acupuncture and Laser Acupuncture for Autonomic Regulations in Rats: Observation on Heart Rate Variability and Gastric Motility 
This study focused on the effects of laser acupuncture (LA) and manual acupuncture (MA) at different acupoints on gastric motility and heart rate variability (HRV) simultaneously to elucidate the site specific effects of acupoints and the correlation between changes of gastric motility and low frequency/high frequency (LF/HF) ratio. Gastric motility and HRV were recorded before and during MA or LA. Stimulating PC-6 or ST-36 significantly enhanced gastric motility, while BL-21 caused no changes. In contrast, MA or LA at CV-12 significantly suppressed gastric motility. Stimulating PC-6 or ST-36 significantly increased heart rate (HR), while CV-12 or BL-21 induced no significant changes of HR. Stimulating PC-6 significantly increased LF/HF, while ST-36, CV-12, or BL-21 induced no significant effects. These results indicated that there was acupoint specificity in the effects of acupuncture on gastric motility and HRV. The stimulatory effect of MA and LA at PC-6 and ST-36 on HR was associated with sympathetic activity. The stimulatory effect of MA or LA at PC-6 or ST-36 on gastric motility was associated with vagal activity. Laser needle can be used as an alternative stimulation therapy.
doi:10.1155/2013/276320
PMCID: PMC3857851  PMID: 24348694
22.  Vaccination with photodynamic therapy-treated macrophages induces highly suppressive T-regulatory cells 
Summary
Background/purpose
The present study explores whether photodynamic therapy (PDT)-induced apoptosis can increase the number of tolerogenic regulatory T cells (Treg) and limit collateral tissue damage.
Methods
BALB/c mice were vaccinated subcutaneously three times with PDT-induced apoptotic or thaw-frozen, necrotic non-infected autologous macrophages (MΦ). Two weeks after the last vaccination, mice were infected intradermally with 106 promastigotes of Leishmania major.
Results
Mice that received PDT-induced apoptotic MΦ had fewer parasites and higher numbers of Treg than mice vaccinated with thaw-frozen necrotic MΦ or phosphate-buffered saline (PBS). Interleukin (IL)-4 and IL-6 were significantly suppressed, while IL-10 was increased in mice that received the PDT-induced apoptotic MΦ. The role of Treg in this process was confirmed through Treg transfer from vaccinated to naïve mice. Mice receiving CD4+CD25+ cells from mice vaccinated with PDT-induced apoptotic MΦ showed smaller lesions 3 weeks after infection and lower parasitic burdens than mice that received Tregs from mice of thaw-frozen necrotic MΦ or PBS groups. These changes were mediated by the depletion of CD3+CD8+ and NKT cells and increased levels of IL-12p70 and interferon-γ, IL-10, and TGF-β in the cutaneous leishmaniasis lesions.
Conclusion
Vaccination with apoptotic MΦ-induced tolerogenic Treg cells that limited collateral tissue damage and diminished parasitic burden.
doi:10.1111/j.1600-0781.2011.00578.x
PMCID: PMC3792570  PMID: 21392113
Leishmania; macrophage; tolerance; T regulatory cells; vaccination
23.  Evaluation of known oncoantibodies, HER2, p53, and cyclin B1, in pre-diagnostic breast cancer sera 
Serum autoantibodies, directed against oncogenic proteins, have been frequently detected in the sera of breast cancer patients. It is unknown whether serum antibodies that are identified in patients with established disease could also be detected in patients with newly diagnosed disease or even predate the diagnosis of breast cancer. Using sera collected at the time of treatment, at the time of diagnosis, or prior to the time of diagnosis, the current study aimed to address the temporal relationship between breast cancer development and serum antibody response. Starting from serum antibodies to eight known breast cancer antigens, we first identified four serum antibodies, HER-2/neu, p53, CEA, and cyclin B1, which are significantly increased in the sera collected from breast cancer patients at the time of treatment. These antibodies were also elevated in breast cancer sera collected at the time of diagnosis. Lastly, comparison of antibody responses in pre-diagnostic samples from women prior to the development of breast cancer and in controls demonstrated that antibodies to the HER-2/neu and p53 can be detected in sera that were collected on average more than 150 days before a breast cancer diagnosis. These results demonstrated that serum autoantibodies commonly reported in sera from patients with established disease can also be detected in pre-diagnostic sera and may be useful for the early detection of breast cancer.
doi:10.1158/1940-6207.CAPR-11-0558
PMCID: PMC3790582  PMID: 22715141
serum antibody; breast cancer; early detection
24.  Amniotic Fluid Stem Cells with Low γ-Interferon Response Showed Behavioral Improvement in Parkinsonism Rat Model 
PLoS ONE  2013;8(9):e76118.
Amniotic fluid stem cells (AFSCs) are multipotent stem cells that may be used in transplantation medicine. In this study, AFSCs established from amniocentesis were characterized on the basis of surface marker expression and differentiation potential. To further investigate the properties of AFSCs for translational applications, we examined the cell surface expression of human leukocyte antigens (HLA) of these cells and estimated the therapeutic effect of AFSCs in parkinsonian rats. The expression profiles of HLA-II and transcription factors were compared between AFSCs and bone marrow-derived mesenchymal stem cells (BMMSCs) following treatment with γ-IFN. We found that stimulation of AFSCs with γ-IFN prompted only a slight increase in the expression of HLA-Ia and HLA-E, and the rare HLA-II expression could also be observed in most AFSCs samples. Consequently, the expression of CIITA and RFX5 was weakly induced by γ-IFN stimulation of AFSCs compared to that of BMMSCs. In the transplantation test, Sprague Dawley rats with 6-hydroxydopamine lesioning of the substantia nigra were used as a parkinsonian-animal model. Following the negative γ-IFN response AFSCs injection, apomorphine-induced rotation was reduced by 75% in AFSCs engrafted parkinsonian rats but was increased by 53% in the control group after 12-weeks post-transplantation. The implanted AFSCs were viable, and were able to migrate into the brain’s circuitry and express specific proteins of dopamine neurons, such as tyrosine hydroxylase and dopamine transporter. In conclusion, the relative insensitivity AFSCs to γ-IFN implies that AFSCs might have immune-tolerance in γ-IFN inflammatory conditions. Furthermore, the effective improvement of AFSCs transplantation for apomorphine-induced rotation paves the way for the clinical application in parkinsonian therapy.
doi:10.1371/journal.pone.0076118
PMCID: PMC3786896  PMID: 24098771
25.  Unusual Features in an Adult Pancreatic Hemangioma: CT and MRI Demonstration 
Korean Journal of Radiology  2013;14(5):781-785.
Hemangiomas in the pancreas are very rare and only a few cases in adulthood have been reported in the literature. We describe a case of pancreatic hemangiomas in an adult with unique imaging findings. A 23-year-old woman visited the hospital for an incidentally detected pancreatic mass. CT and MRI revealed a multilocular cyst with fluid-fluid levels and no obvious enhancement. The patient underwent surgery and the mass was confirmed as a pancreatic hemangioma. The radiological features and differential diagnosis of this rare lesion are discussed.
doi:10.3348/kjr.2013.14.5.781
PMCID: PMC3772258  PMID: 24043972
Pancreas; Hemangioma; Computed tomography; Magnetic resonance imaging

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