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Articles by Willey Liao in JoVE

 JoVE Biology

A Novel Bayesian Change-point Algorithm for Genome-wide Analysis of Diverse ChIPseq Data Types

1Department of Applied Mathematics & Statistics, Stony Brook University, 2Computational Biology and Bioinformatics, Cold Spring Harbor Laboratory, 3Department of Molecular and Cell Biology, University of Texas at Dallas


JoVE 4273

Our Bayesian Change Point (BCP) algorithm builds on state-of-the-art advances in modeling change-points via Hidden Markov Models and applies them to chromatin immunoprecipitation sequencing (ChIPseq) data analysis. BCP performs well in both broad and punctate data types, but excels in accurately identifying robust, reproducible islands of diffuse histone enrichment.

Other articles by Willey Liao on PubMed

Vaspin Attenuates the Apoptosis of Human Osteoblasts Through ERK Signaling Pathway

It has been hypothesized that adipocytokines originating from adipose tissue may have an important role in bone metabolism. Vaspin is a novel adipocytokine isolated from visceral white adipose tissue, which has been reported to have anti-apoptotic effects in vascular endothelial cells. However, to the best of our knowledge there is no information regarding the effects of vaspin on osteoblast apoptosis. This study therefore examined the possible effects of vaspin on apoptosis in human osteoblasts (hOBs). Our study established that vaspin inhibits hOBs apoptosis induced by serum deprivation, as determined by ELISA and TUNEL assays. Western blot analysis revealed that vaspin upregulates the expression of Bcl-2 and downregulates that of Bax in a dose-dependent manner. Vaspin stimulated the phosphorylation of ERK, and pretreatment of hOBs with the ERK inhibitor PD98059 blocked the vaspin-induced activation of ERK, however, vaspin did not stimulate the phosphorylation of p38, JNK or Akt. Vaspin protects hOBs from serum deprivation-induced apoptosis, which may be mediated by activating the MAPK/ERK signaling pathway.

Novel Foxo1-dependent Transcriptional Programs Control T(reg) Cell Function

Regulatory T (T(reg)) cells, characterized by expression of the transcription factor forkhead box P3 (Foxp3), maintain immune homeostasis by suppressing self-destructive immune responses. Foxp3 operates as a late-acting differentiation factor controlling T(reg) cell homeostasis and function, whereas the early T(reg)-cell-lineage commitment is regulated by the Akt kinase and the forkhead box O (Foxo) family of transcription factors. However, whether Foxo proteins act beyond the T(reg)-cell-commitment stage to control T(reg) cell homeostasis and function remains largely unexplored. Here we show that Foxo1 is a pivotal regulator of T(reg )cell function. T(reg) cells express high amounts of Foxo1 and display reduced T-cell-receptor-induced Akt activation, Foxo1 phosphorylation and Foxo1 nuclear exclusion. Mice with T(reg)-cell-specific deletion of Foxo1 develop a fatal inflammatory disorder similar in severity to that seen in Foxp3-deficient mice, but without the loss of T(reg) cells. Genome-wide analysis of Foxo1 binding sites reveals ~300 Foxo1-bound target genes, including the pro-inflammatory cytokine Ifng, that do not seem to be directly regulated by Foxp3. These findings show that the evolutionarily ancient Akt-Foxo1 signalling module controls a novel genetic program indispensable for T(reg) cell function.

Hyperthermia Inhibits the Proliferation and Invasive Ability of Mouse Malignant Melanoma Through TGF-β1

The degradation of basement membranes by tumor cells involves secretion and activation of proteinases, such as the matrix metalloproteinases (MMPs), and results from an imbalance between their inhibitors and activators that are controlled by various growth factors or cytokines, among which TGF-β1 may be the most intriguing. In order to study the therapeutic effect and molecular mechanism of hyperthermia on aggressive malignant melanoma, the expression levels of TGF-β1 and Smad4 in B16F10 cells were dynamically analyzed by RT-PCR and western blotting for 24 h after heat treatment, from which time-dependent changes were determined. As expected, the proliferation and invasive ability of B16F10 cells were suppressed strongly by heat treatment. Furthermore, we compared the expression of TGF-β1 in melanoma mouse models before and after magnetic fluid hyperthermia (MFH) in vivo. After hyperthermia, the tumor growth rate was reduced with a decline in TGF-β1 protein expression. We conclude that changes in the TGF-β1 pathway induced by hyperthermia may be an important part of the molecular mechanism involved.

An Integrated Genome-wide Approach to Discover Tumor Specific Antigens As Potential Immunological and Clinical Targets in Cancer

Tumor-specific antigens (TSAs) are central elements in the immune control of cancers. To systematically explore the TSA genome, we developed a computational technology called Heterogeneous Expression Profile Analysis (HEPA), which can identify genes relatively uniquely expressed in cancer cells in contrast to normal somatic tissues. Rating human genes by their HEPA score enriched for clinically useful TSA genes, nominating candidate targets whose tumor-specific expression was verified by RT-PCR. Coupled with HEPA, we designed a novel assay termed Protein A/G based Reverse Serological Evaluation (PARSE) for quick detection of serum autoantibodies against an array of putative TSA genes. Remarkably, highly tumor-specific autoantibody responses against seven candidate targets were detected in 4-11% of patients, resulting in distinctive autoantibody signatures in lung and stomach cancers. Interrogation of a larger cohort of 149 patients and 123 healthy individuals validated the predictive value of the autoantibody signature for lung cancer. Together, our results establish an integrated technology to uncover a cancer-specific antigen genome offering a reservoir of novel immunological and clinical targets.

Abcg2 Regulates Cell Cycle Progression and Asymmetric Division in Mouse Cardiac Side Population Progenitor Cells

Rationale: Following cardiac injury, cardiac progenitor cells are acutely reduced, and are replenished in part by regulated self-renewal and proliferation, which occurs through symmetric and asymmetric cellular division. Understanding the molecular cues controlling progenitor cell self-renewal and lineage commitment is critical towards harnessing these cells for therapeutic regeneration. We have previously found that the cell surface ATP binding cassette (ABC)-transporter, Abcg2, influences the proliferation of cardiac side population (CSP) progenitor cells, though through unclear mechanisms. Objective: To determine the role of Abcg2 on cell cycle progression and mode of division in mouse CSP cells. Methods and Results: Herein, using CSP cells isolated from wild-type and Abcg2-knockout mice, we find that Abcg2 regulates G1-S cell cycle transition by FUCCI cell cycle indicators, cell cycle-focused gene expression arrays and confocal live cell fluorescent microscopy. Moreover, we find that modulation of cell cycle results in transition from symmetric to asymmetric cellular division in CSP cells lacking Abcg2. Conclusions: Abcg2 modulates CSP cell cycle progression and asymmetric cell division, establishing a mechanistic link between this surface transporter and cardiac progenitor cell function. Greater understanding of progenitor cell biology, and in particular the regulation of resident progenitor cell homeostasis, is vital for guiding the future development of cell-based therapies for cardiac regeneration.

Amaryllidaceae Alkaloids Inhibit Nuclear-to-cytoplasmic Export of Ribonucleoprotein (RNP) Complex of Highly Pathogenic Avian Influenza Virus H5N1

Please cite this paper as: He et al. (2012) Amaryllidaceae alkaloids inhibit nuclear-to-cytoplasmic export of ribonucleoprotein (RNP) complex of highly pathogenic avian influenza virus H5N1. Influenza and Other Respiratory Viruses DOI: 10.1111/irv.12035. Background  Few drugs are currently licensed to treat influenza A infection, and new therapies are needed, especially for highly pathogenic strains. Traditional medicinal plants, such as Lycoris radiata, are a potential source of new antiviral agents. Objective  To test 15 Amaryllidaceae alkaloids isolated from the bulbs of L. radiata in vitro for antiviral activities against influenza virus type A, A/Chicken/GuangDong/178/2004 (H5N1, 178). Methods  Antiviral activities of the compounds were tested in time-of-addition assays, hemagglutination inhibition (HI) assays, neuraminidase (NA) activity assays, and viral entry inhibition assays using H5N1-HIV pseudoviruses. Effects of the compounds on localization and activity of the viral ribonucleoprotein (RNP) were determined by immunofluorescence and an RNP minigenome assay, respectively. Results  Among the alkaloids, lycorine (AA1), hippeastrine (AA2), hemanthamine (AA3) and 11-hydroxy vittatine (AA4) exhibited antiviral activities, with EC(90) values of 0·52, 82·07, 4·15, and 13·45 μm, respectively. These compounds did not affect the function of the outer membrane proteins or the viral entry process and viral RNP activity. As AA1 and AA3 exhibited stronger antiviral activities, they were further analyzed. Intracellular nucleoprotein (NP) localization showed that AA1 and AA3 inhibited the RNP complex in the nucleus at an early stage of a single-round and multi-round of replication. Conclusion  Four Amaryllidaceae alkaloids were first determined that could exert anti-influenza activities after virus entry into cells. Furthermore, AA1 and AA3 could inhibit nuclear-to-cytoplasmic export of the RNP complex of virus replication. Thus, these compounds may be developed further as anti-influenza drug candidates.

Overexpression of Chemokine Ligand 7 is Associated with the Progression of Canine Transmissible Venereal Tumor

ABSTRACT: BACKGROUND: Chemokines play multiple roles in the development and progression in a variety of tumors. Chemokine (C-X-C motif) ligand 7 (CXCL7) has been found associated with pro-inflammatory responses, but its role in cancer growth remains unclear. Our previous study showed that R phase tumor infiltrating lymphocytes (TILs) produced large amounts of interleukin (IL)-6 which antagonized transforming growth factor (TGF)-beta derived from CTVT to diminish the immune-suppressive microenvironment. Now we intend to determine the expression pattern of CXCL7 and the role of IL-6/TGF-beta in CXCL7 induction during spontaneous progressive (P) and regressive (R) phases in canine transmissible venereal tumor (CTVT). RESULTS: We have demonstrated that CXCL7 expressed at high level in P phase and down-regulated in R phase by western blot and real-time PCR. This suggested that CXCL7 expression was negatively correlated with the tumor growth. Co-culturing TILs with CTVT cells was found to reduce CXCL7 expression, while adding IL-6 blocking antibody reversed it. Moreover, in P phase CTVT, while IL-1beta and TGF-beta had no obvious effect on CXCL7 expression, IL-6 was found significantly to reduce CXCL7 expression in a dose-dependent manner. The mRNA expression results of CXCL7 receptor, CXCR2, further confirmed the effects of IL-6 concentration on the CXCL7 expression. CONCLUSION: CXCL7 overexpression might be associated with the progressive growth of CTVT. The results shown here also suggest the role of CXCL7 in cancer development and the potential as the anti-cancer therapeutic target.

Novel Susceptibility Genes Associated with Diabetic Cataract in a Taiwanese Population

Purpose: To identify genetic variants that predispose to type 2 diabetes (T2D) with cataract. Patients and methods: Genome-wide association study (GWAS) of T2D patients with cataract, as graded by Lens Opacities Classification System (LOCS). A total of 109 T2D patients with cataract score equal to or above 10 designated as the study group, 649 T2D patients with cataract score equal to or below 3 as the control group. Single nucleotide polymorphisms (SNPs) with p-values < 10(-5) were considered to be putatively associated with the diabetic cataract. Results: Fifteen SNPs were found to be putatively associated with diabetic cataract. These variants were located near the following genes: PPARD, CCDC102A, GBA3, NEDD9, GABRR1/2, RPS6KA2, tcag7.1163, TAC1, GALNTL1 and KIAA1671. We defined haplotype 1 to haplotype 4 from the alternative alleles of related polymorphisms. Distribution of haplotype 2 on chromosome 4 and haplotype 4 on chromosome 7 revealed significant differences (OR = 1.86 and 1.69, respectively; 95% confidence interval were 1.26-2.76 and 1.23-2.31, respectively). Conclusions: The 15 loci coded on chromosomes 4, 6, 7, 14, 16 and 22 were associated with diabetic cataract. Gene functions are either with mechanisms of regulating blood sugar or formation of cataract. High linkage disequilibrium appeared on chromosome 4p15.31 and chromosome 7q21.3.

GsmR, a Response Regulator with an HD-related Output Domain in Xanthomonas campestris, is Positively Controlled by Clp and is Involved in the Expression of Genes Responsible for Flagellum Synthesis

In prokaryotes, two-component signal transduction systems, consisting of a histidine kinase and a response regulator, play a critical role in regulating a range of cellular functions. A recent study suggests that XCC3315, a response regulator with a CheY-like receiver domain attached to an uncharacterized HD-related output domain (HDOD domain), plays a role in the general stress response of the Gram-negative bacterium Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot in cruciferous plants. Here, we demonstrated genetically that XCC3315, designated as gsmR (general stress and motility regulator), is involved in the expression of genes responsible for flagellum synthesis, including rpoN2, flhF, flhB, and fliC. Site-directed mutagenesis revealed that Glu9 and Arg100 in the receiver domain and Gly205, Asp263, His287, Trp298 and His311 in the HDOD are critical amino acids for GsmR function in cell motility regulation. The gsmR transcription initiation site was mapped. Promoter analysis and gel retardation assay revealed that the expression of gsmR is positively controlled by the global transcriptional regulator Clp in a direct manner, and is subject to catabolite repression. Our findings not only extend the previous work on Clp regulation to show that it influences the expression of gsmR in Xcc, but are also the first to characterize the expression of this response regulator gene in this phytopathogen. Furthermore, GsmR is the first HDOD-containing protein of bacteria in which key amino acids have been experimentally identified and characterized.

Obscure Hemosuccus Pancreaticus Due to Dorsal Pancreatic Arteriorrhexis

Clinical Application of Massively Parallel Sequencing-based Prenatal Noninvasive Fetal Trisomy Test for Trisomies 21 and 18 in 11 105 Pregnancies with Mixed Risk Factors

OBJECTIVE: To report the performance of massively parallel sequencing (MPS) based prenatal noninvasive fetal trisomy test based on cell-free DNA sequencing from maternal plasma in a routine clinical setting in China. METHOD: The MPS-based test was offered as a prenatal screening test for trisomies 21 and 18 to pregnant women in 49 medical centers over 2 years. A total of 11 263 participants were recruited and the MPS-based test was performed in 11 105 pregnancies. Fetal outcome data were obtained after the expected date of confinement. RESULTS: One hundred ninety cases were classified as positive, including 143 cases of trisomy 21 and 47 cases of trisomy 18. With the karyotyping results and the feedback of fetal outcome data, we observed one false positive case of trisomy 21, one false positive case of trisomy 18 and no false negative cases, indicating 100% sensitivity and 99.96% specificity for the detection of trisomies 21 and 18. CONCLUSION: Our large-scale multicenter study proved that the MPS-based test is of high sensitivity and specificity in detecting fetal trisomies 21 and 18. The introduction of this screening test into a routine clinical setting could avoid about 98% of invasive prenatal diagnostic procedures. © 2012 John Wiley & Sons, Ltd.

Penehyclidine Hydrochloride: a Potential Drug for Treating COPD by Attenuating Toll-like Receptors

The aim of this review was to evaluate and summarize the available scientific information on penehyclidine hydrochloride (PHC) for the treatment of chronic obstructive pulmonary disease (COPD) as a result of its ability to attenuate Toll-like receptors. Penehyclidine hydrochloride is an anticholinergic drug manufactured in China, with both antimuscarinic and antinicotinic activity. PHC is used widely in the clinic as a reversal agent in cases of organic phosphorus poisoning and soman poisoning, but also may also have an important role as a bronchodilator in the treatment of obstructive airway disease, including asthma and, in particular, COPD.

Folliculin, the Product of the Birt-Hogg-Dube Tumor Suppressor Gene, Interacts with the Adherens Junction Protein P0071 to Regulate Cell-Cell Adhesion

Birt-Hogg-Dube (BHD) is a tumor suppressor gene syndrome associated with fibrofolliculomas, cystic lung disease, and chromophobe renal cell carcinoma. In seeking to elucidate the pathogenesis of BHD, we discovered a physical interaction between folliculin (FLCN), the protein product of the BHD gene, and p0071, an armadillo repeat containing protein that localizes to the cytoplasm and to adherens junctions. Adherens junctions are one of the three cell-cell junctions that are essential to the establishment and maintenance of the cellular architecture of all epithelial tissues. Surprisingly, we found that downregulation of FLCN leads to increased cell-cell adhesion in functional cell-based assays and disruption of cell polarity in a three-dimensional lumen-forming assay, both of which are phenocopied by downregulation of p0071. These data indicate that the FLCN-p0071 protein complex is a negative regulator of cell-cell adhesion. We also found that FLCN positively regulates RhoA activity and Rho-associated kinase activity, consistent with the only known function of p0071. Finally, to examine the role of Flcn loss on cell-cell adhesion in vivo, we utilized keratin-14 cre-recombinase (K14-cre) to inactivate Flcn in the mouse epidermis. The K14-Cre-Bhd(flox/flox) mice have striking delays in eyelid opening, wavy fur, hair loss, and epidermal hyperplasia with increased levels of mammalian target of rapamycin complex 1 (mTORC1) activity. These data support a model in which dysregulation of the FLCN-p0071 interaction leads to alterations in cell adhesion, cell polarity, and RhoA signaling, with broad implications for the role of cell-cell adhesion molecules in the pathogenesis of human disease, including emphysema and renal cell carcinoma.

L-Carnitine Is an Endogenous HDAC Inhibitor Selectively Inhibiting Cancer Cell Growth In Vivo and In Vitro

L-carnitine (LC) is generally believed to transport long-chain acyl groups from fatty acids into the mitochondrial matrix for ATP generation via the citric acid cycle. Based on Warburg's theory that most cancer cells mainly depend on glycolysis for ATP generation, we hypothesize that, LC treatment would lead to disturbance of cellular metabolism and cytotoxicity in cancer cells. In this study, Human hepatoma HepG2, SMMC-7721 cell lines, primary cultured thymocytes and mice bearing HepG2 tumor were used. ATP content was detected by HPLC assay. Cell cycle, cell death and cell viability were assayed by flow cytometry and MTS respectively. Gene, mRNA expression and protein level were detected by gene microarray, Real-time PCR and Western blot respectively. HDAC activities and histone acetylation were detected both in test tube and in cultured cells. A molecular docking study was carried out with CDOCKER protocol of Discovery Studio 2.0 to predict the molecular interaction between L-carnitine and HDAC. Here we found that (1) LC treatment selectively inhibited cancer cell growth in vivo and in vitro; (2) LC treatment selectively induces the expression of p21(cip1) gene, mRNA and protein in cancer cells but not p27(kip1); (4) LC increases histone acetylation and induces accumulation of acetylated histones both in normal thymocytes and cancer cells; (5) LC directly inhibits HDAC I/II activities via binding to the active sites of HDAC and induces histone acetylation and lysine-acetylation accumulation in vitro; (6) LC treatment induces accumulation of acetylated histones in chromatin associated with the p21(cip1) gene but not p27(kip1) detected by ChIP assay. These data support that LC, besides transporting acyl group, works as an endogenous HDAC inhibitor in the cell, which would be of physiological and pathological importance.

Loss of the Respiratory Enzyme Citrate Synthase Directly Links the Warburg Effect to Tumor Malignancy

To investigate whether altered energy metabolism induces the Warburg effect and results in tumor malignancy, the respiratory enzyme citrate synthase (CS) was examined, silenced, and the effects analyzed. In human cervical carcinoma cells, RNAi-mediated CS knockdown induced morphological changes characteristic of the epithelial-mesenchymal transition (EMT). This switch accelerated cancer cell metastasis and proliferation in in vitro assays and in vivo tumor xenograft models. Notably, CS knockdown cells exhibited severe defects in respiratory activity and marked decreases in ATP production, but great increases in glycolytic metabolism. This malignant progression was due to activation of EMT-related regulators; altered energy metabolism resulted from deregulation of the p53/TIGAR and SCO2 pathways. This phenotypic change was completely reversed by p53 reactivation via treatment with proteasome inhibitor MG132 or co-knockdown of E3 ligase HDM2 and partially suppressed by ATP treatment. This study directly links the Warburg effect to tumor malignancy via induction of the EMT phenotype.

Regenerative Potential of Decellularized Porcine Nucleus Pulposus Hydrogel Scaffolds; Stem Cell Differentiation, Matrix Remodeling and Biocompatibility Studies

Nucleus pulposus (NP) tissue regeneration has been proposed as an early-stage interventional therapy to combat intervertebral disc degeneration. We have previously reported on the development and characterization of a novel biomimetic acellular porcine nucleus pulposus (APNP) hydrogel. Herein we aimed to evaluate this material for use as a suitable scaffold for NP tissue regeneration. Human adipose derived stem cells (hADSCs) were cultured for 14 days on APNP hydrogels in chemically defined differentiation media and were analyzed for an NP cell-like mRNA expression profile, evidence of hydrogel remodeling including hydrogel contraction measurements, extracellular matrix production and compressive dynamic mechanical properties. The innate capacity of the hydrogel itself to induce stem cell differentiation was also examined via culture in media lacking soluble differentiation factors. Additionally, the in vivo biocompatibility of non-crosslinked and ethyldimethylaminopropyl carbodiimide/ N-hydroxysuccinimide (EDC/NHS) and pentagalloyl glucose (PGG) crosslinked hydrogels was evaluated in a rat subdermal model. Results indicated that hADSCs expressed putative NP cell positive gene transcript markers when cultured on APNP hydrogels. Additionally, glycosaminoglycan and collagen content of hADSC seeded hydrogels was significantly greater than non-seeded controls and cell seeded hydrogels exhibited evidence of contraction and TIMP-1 production. The dynamic mechanical properties of the hADSC seeded hydrogels increased with time in culture in comparison to non cell-seeded controls and approached values reported for native NP tissue. Immunohistochemical analysis of explants illustrated the presence of mononuclear cells including macrophages and fibroblasts as well as blood vessel infiltration and collagen deposition within the implant interstices after 4 weeks of implantation. Taken together, these results suggest that APNP hydrogels, in concert with autologous ADSCs, may serve as a suitable scaffold for NP tissue regeneration.

Tissue-engineered Tubular Graft for Urinary Diversion After Radical Cystectomy in Rabbits

BACKGROUND: Clinically, using ileal conduit for urinary diversion often caused many serious complications. Tissue engineering technology may offer an alternative method for urinary diversion after radical cystectomy. In this study, we aimed to make a tissue-engineered tubular graft (TETG) using bladder epithelial cells and bladder acellular matrix (BAM) for urinary diversion in rabbits. METHODS: Bladder epithelial cells of rabbit were cultivated and expanded in vitro, which were then seeded on BAM and cultured for 7 d. Then, cell-seeded grafts of 4 cm length and 0.8 cm diameter were used to make TETGs and transferred into the omentum for 2 wk before urinary diversion. In the experimental group, bladders of the rabbits were removed. The proximal ends of TETGs were anastomosed with ureters, and the distal ends were anastomosed with the abdominal stomas. In the control group, TETGs were constructed using unseeded BAM. Newly formed tissue structures were functionally and microscopically evaluated using urography and immunohistochemistry at 1, 2, 4, and 8 wk after operation, respectively. Histologic examination with hematoxylin and eosin staining was conducted to assess tissue regeneration. Immunohistochemistry was performed with AE(1)/AE(3), uroplakin Ⅲa, and zonula occludens 1 (ZO-1) antibodies. RESULTS: All animals were alive in the experimental group. Hematoxylin and eosin staining showed epithelial coverage in TETG. Immunohistochemistry showed positive stain with AE(1)/AE(3), uroplakin Ⅲa, and ZO-1, which indicated mature and functional epithelial cells on the lumen of TETG. Intravenous urography showed that there were no obstructions in TETGs. In the control group, four rabbits were dead within 2 wk, and scar formation, atresia, and severe hydronephrosis were found. CONCLUSIONS: It was feasible that TETG constructed using bladder epithelial cells and BAM for urinary diversion after radical cystectomy in rabbits.

A Metabonomic Approach to the Effect Evaluation of Treatment in Patients Infected with Influenza A (H1N1)

The pandemic influenza A virus (H1N1) was transmitted to the human population since 2009, resulting in some consequences of viral pneumonia, respiratory failure, multiple organ failure and, most severely, death. In clinical practice, Chinese medicine possessed extensive experience for prevention and treatment of influenza, but its mechanism still remain unclear. In addition, the efficacy of combination therapy of Chinese and Western medicine was attractive, but not yet clear. In the present study, 131 patients from Guangzhou China referred for H1N1 virus mRNA testing for the evaluation of possible influenza A-infected were eligible for participation. A metabonomics study was carried out to explore the difference between before and after treatment in patients with H1N1 through Chinese and/or Western medicine. Results from metabolic profiling and biochemical detection indicated significant metabolic change in the arachidonic acid metabolic pathway. In the group of combination therapy of Chinese and western medicine, its efficacy was best and the potential biomarkers were significantly changed compared with untreated state. Those results indicated that the potential metabolic biomarker could be supplemented with biochemical detection to obtain more precise diagnosis for H1N1 infection. Clinical trials registration: Clinical Trials. Gov No. 2008GL-50.

Relieving Visceral Hyperalgesia Effect of Kangtai Capsule and Its Potential Mechanisms Via Modulating the 5-HT and NO Level in Vivo

Kangtai capsule (KT) is one type of traditional Chinese medicine preparation derived from the proved recipe, which was frequently applied as an effective clinical treatment of IBS. However, there still lack the reasonable and all-round analytical approach and the scientific studies on its underlying mechanisms. Therefore, our study aimed to develop the novel method for evaluating its quality as well as to interpret the potential mechanisms. In our study, high performance liquid chromatography (HPLC) fingerprint was applied to provide a chemical profile of KT. The neonatal maternal separation (NMS) on Sprague-Dawley pups was employed to evaluate the therapeutic effect of KT by virtue of various parameters including visceral hyperalgesia, serum nitric oxide (NO) level, and tissue 5-hydroxytryptamine (5-HT) level. Consequently, a chromatographic condition, which was carried at 30°C with a flow rate of 0.5ml/min on AQUA 3μ C18 column with mobile phase of acetonitrile and water-phosphoric acid (100:0.1, v/v), was established to give a common fingerprint chromatography under 254nm with a similarity index of 0.963 within ten batches of KT samples. On the NMS model, KT markedly elevated the pain threshold of NMS rats. Furthermore, KT at three doses significantly decreased 5-HT content from distal colon of visceral hyperalgesia rats induced by NMS, while the significant decrease of 5-HT content in serum was only observed in the group with KT at high dose. However, compared with that in NMS rats without KT, there was no apparent difference of 5-HT level from brain issue in the rats with various doses. Besides, KT could substantially elevate the concentration of NO in the serum. The results showed our study developed the simple, rapid, accurate, reproducible qualitative and quantitative analysis by HPLC fingerprint for the quality control for KT. Data from the pharmacological investigation suggested that the curative effect of KT to the visceral hypersensitivity may be concerned with the level of 5-HT and NO in vivo, promising its potential in irritable bowel syndrome treatment.

HDAC4 Governs a Transcriptional Program Essential for Synaptic Plasticity and Memory

Neuronal activity influences genes involved in circuit development and information processing. However, the molecular basis of this process remains poorly understood. We found that HDAC4, a histone deacetylase that shuttles between the nucleus and cytoplasm, controls a transcriptional program essential for synaptic plasticity and memory. The nuclear import of HDAC4 and its association with chromatin is negatively regulated by NMDA receptors. In the nucleus, HDAC4 represses genes encoding constituents of central synapses, thereby affecting synaptic architecture and strength. Furthermore, we show that a truncated form of HDAC4 encoded by an allele associated with mental retardation is a gain-of-function nuclear repressor that abolishes transcription and synaptic transmission despite the loss of the deacetylase domain. Accordingly, mice carrying a mutant that mimics this allele exhibit deficits in neurotransmission, spatial learning, and memory. These studies elucidate a mechanism of experience-dependent plasticity and define the biological role of HDAC4 in the brain.

A Possible Correlation Between Oxytocin-induced and Angiotensin IV-induced Anti-hyperalgesia at the Spinal Level in Rats

In our previous study, we showed that intrathecal (i.t.) administration of angiotensin IV (Ang IV), an insulin-regulated aminopeptidase (IRAP) inhibitor, attenuated inflammatory hyperalgesia in rats. Using the plantar test in rats with carrageenan-induced paw inflammation, we investigated the possible mechanism(s) of this effect. Because i.t. oxytocin was reported to produce a dose-dependent anti-hyperalgesia in rats with inflammation, we speculate that there is a possible correlation between oxytocin-induced and Ang IV-induced anti-hyperalgesia. Using i.t. co-administered atosiban (oxytocin receptor antagonist), the anti-hyperalgesia by Ang IV was completely abolished. This indicated that oxytocin could be the major IRAP substrate responsible for the anti-hyperalgesia by Ang IV. When Ang IV was co-administered with a low dose of oxytocin, there was a significant enhancing effect of Ang IV on oxytocin-induced anti-hyperalgesia. In recent reports, electrical stimulation on the paraventricular hypothalamic nucleus (PVN) was proved to increase oxytocin release at the spinal cord. Our results also showed that Ang IV could prolong the anti-hyperalgesia induced by PVN stimulation. This suggests a possible protective effect of Ang IV on endogenous oxytocin degradation/dysfunctioning. Moreover, we examined the local effect of intraplantarly injected Ang IV in the same model. Our results showed no effect of local Ang IV on hyperalgesia and paw edema, indicating that Ang IV may not regulate the peripheral inflammatory process. Overall, our study suggests that Ang IV may act through the inhibition of the activity of IRAP to reduce the degradation of oxytocin at the spinal cord, thereby leading to anti-hyperalgesia in rats with inflammation.

MicroRNA-29c Enhances the Sensitivities of Human Nasopharyngeal Carcinoma to Cisplatin-based Chemotherapy and Radiotherapy

This study was aimed to investigate the potential role of microRNA-29c (miR-29c) in regulating the sensitivities of nasopharyngeal carcinoma (NPC) to ionizing radiation (IR) and cisplatin. Low expression of miR-29c was positively associated with therapeutic resistance in 159 NPC cases. Our further in vitro and in vivo studies illustrated ectopic restoration of miR-29c substantially enhanced the sensitivity of NPC cells to IR and cisplatin treatment by promoting apoptosis. Furthermore, we detected miR-29c repressed expression of anti-apoptotic factors, Mcl-1 and Bcl-2 in NPC tissues and cell lines. These data indicate miR-29c might serve as a potential therapeutic sensitizer in NPC treatment.

Protective Effects of Erythropoietin on Astrocytic Swelling After Oxygen-glucose Deprivation and Reoxygenation: Mediation Through AQP4 Expression and MAPK Pathway

Recent in vivo studies have shown that erythropoietin (EPO) offers strong protection against brain edema. However, the intracellular and molecular mechanisms behind this beneficial effect have not been specified. The aim of this study was to determine whether human erythropoietin (rhEPO) reduces the astrocytic swelling created by oxygen-glucose deprivation followed by reoxygenation (OGD/Reox) in vitro and whether this effect can be mediated through the modulation of aquaporin4 (AQP4) expression in the plasma membrane (PM) and phosphorylation of the mitogen-activated protein kinase pathway (MAPK) pathway. Our results showed that OGD/Reox produced increase in cell volume, morphological swelling, and mitochondrial swelling. These changes were associated with the up-regulation of AQP4 in PM and the over-activation of MAPK. Silencing AQP4 expression using small interfering ribonucleic acid for AQP4 was found to block astrocytic swelling. Inhibition of the over-activation of MAPK mitigated the PM AQP4 overabundance and cellular swelling. As expected, treatment with rhEPO significantly reduced the OGD/Reox-induced increase in cell volume, morphological swelling, and mitochondrial swelling as well as the up-regulation of AQP4 in PM. In addition, cultures treated with the neutralizing anti-EPO antibody worsened the PM AQP4 abundance and cellular swelling, abolishing the protective effects mediated by rhEPO treatment. Furthermore, the over-activation of these MAPK after OGD/Reox was attenuated by rhEPO treatment significantly. In conclusion, our data strongly suggest that rhEPO can protect astrocytes from swelling caused by ischemia and reperfusion-like injury. This neuroprotective capacity is partially mediated by diminishing the MAPK-activity-dependent overabundance of AQP4 in PM.

Central Venous Oxygen Saturation Under Non-Protocolized Resuscitation Is Not Related to Survival in Severe Sepsis or Septic Shock

ABSTRACT: Protocolized hemodynamic resuscitation in severe sepsis or septic shock is not universally applied in all emergency departments and general hospital wards around the world. It is unknown whether ScvO2 levels are associated with the clinical outcome of severe sepsis or septic shock under nonprotocolized resuscitation. In this prospective study, we enrolled 124 noncirrhotic patients who were admitted to intensive care units for severe sepsis or septic shock. The average Acute Physiology and Chronic Health Evaluation II score was 25.3 (SD, 7.6). According to ScvO2 levels after initial resuscitation before intensive care unit admission, patients were divided into high (ScvO2 ≥ 70%, n = 63) and low (ScvO2 < 70%, n = 61) ScvO2 groups. Compared with high ScvO2 groups, low ScvO2 groups showed no significant differences in 28-day mortality (25.4% vs. 24.6%; P = 0.943) or hospital mortality (30.2% vs. 31.1%; P = 0.794). Multivariate logistic regression models showed that low mean arterial pressure (hazard ratio, 0.967; 95% confidence interval, 0.940-0.994; P = 0.019) and high central venous pressure (hazard ratio, 1.150; 95% confidence interval, 1.057-1.251; P = 0.001) after initial resuscitation were associated with higher 28-day mortality. On the contrary, ScvO2 levels after resuscitation were not related to 28-day or hospital mortality. In conclusion, our results showed that mean arterial pressure and central venous pressure were still the most important hemodynamic variables in initial hemodynamic resuscitation. Low postresuscitation ScvO2 was not associated with a worse outcome. It is possible that ScvO2 less than 70% might not necessarily be associated with tissue hypoxia, and critical ScvO2 levels require to be determined by further studies.

BMP-2, VEGF and BFGF Synergistically Promote the Osteogenic Differentiation of Rat Bone Marrow-derived Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) were treated with bone morphogenetic protein-2 (BMP-2), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) dose-dependently and time-dependently. Together they caused a strong synergistic effect on the osteogenic differentiation of MSCs, with lower concentrations of each factor being enough to show the synergistic promotion (50 ng BMP-2/ml, 1 ng VEGF/ml and 10 ng bFGF/ml). When both VEGF and bFGF were added in the early proliferating stage (the first 7 days) and BMP-2 was added in the late differentiation stage (the last 7 days), osteogenic differentiation of MSCs could be enhanced more effectively.

Identification of 15 New Psoriasis Susceptibility Loci Highlights the Role of Innate Immunity

To gain further insight into the genetic architecture of psoriasis, we conducted a meta-analysis of 3 genome-wide association studies (GWAS) and 2 independent data sets genotyped on the Immunochip, including 10,588 cases and 22,806 controls. We identified 15 new susceptibility loci, increasing to 36 the number associated with psoriasis in European individuals. We also identified, using conditional analyses, five independent signals within previously known loci. The newly identified loci shared with other autoimmune diseases include candidate genes with roles in regulating T-cell function (such as RUNX3, TAGAP and STAT3). Notably, they included candidate genes whose products are involved in innate host defense, including interferon-mediated antiviral responses (DDX58), macrophage activation (ZC3H12C) and nuclear factor (NF)-κB signaling (CARD14 and CARM1). These results portend a better understanding of shared and distinctive genetic determinants of immune-mediated inflammatory disorders and emphasize the importance of the skin in innate and acquired host defense.

Role of Pharmacogenetics on Adjuvant Chemotherapy-induced Neutropenia in Chinese Breast Cancer Patients

BACKGROUND: Breast cancer patients regularly undergo adjuvant chemotherapies following surgery. However, these treatments are largely associated with chemotherapeutic toxicities ranging from nausea to severe myelosuppression. In this investigation, we examined the effects of four SNPs in NR1I2, CYP3A4 and CYP3A5 genes on chemotherapy-induced severe neutropenia in 311 female Chinese breast cancer patients undergoing a standard adjuvant chemotherapy regimen. METHODS: Patients were monitored for adverse reactions throughout the treatment, then divided into "none or mild" (80 %) or "severe" (20 %) toxicity groups according to whether they suffered grade 4 neutropenia defined as having an absolute neutrophil counts (ANC) of less than 0.5 × 10(9)/L anytime during the treatment. DNA was extracted from patients' peripheral blood samples, then genotyped using allele-specific Tm-shift PCR and melting analysis. RESULTS: Logistic regression revealed that rs776746 or CYP3A5*3 strongly associated with grade 4 neutropenia (OR = 2.56, P = 0.023) after adjustment for covariates, one of which more significant factor was baseline ANC (OR = 0.68, P = 0.020). Although univariate analysis in all patients did not reveal any association at first, further analysis indicated that rs776746 is significantly associated with severe neutropenia in subgroup of breast cancer patients with normal baseline ANC (≥2.0 × 10(9)/L). These carriers of A-allele have 3.14-fold increased risk of developing severe neutropenia (P = 0.004). CONCLUSION: Our results suggested that polymorphisms in CYP3A5 might be useful pharmacogenetic markers for the prediction of severe neutropenia during chemotherapy, however, only after screening patients by their baseline ANC in the presence of gene-environmental interaction. We demonstrate an approach of pharmacogenetic analysis, in which the genetic data should be analyzed in the perspective of other clinical parameters.

Comparative Interface Metrics for Metal-Free Monolayer-Based Dye-Sensitized Solar Cells

The first quantitative comparison between self-assembled monolayers of homologous carboxylate- and phosphonate-terminated organic dyes that are of use in dye-sensitized solar cells (DSSCs) is reported. (Cyanovinyl)phosphonate-terminated oligothiophenes and (cyanovinyl)carboxylate-terminated oligothiophenes were synthesized on TiO(2) thin film electrodes. Structurally analogous organics were compared for the effect of the anchoring groups on photochemical properties in solution as measured by UV/vis spectroscopy and for reactivity with the electrode surface. Monolayers were grown on the TiO(2) electrodes either by "tethering by aggregation and growth" (T-BAG) or by solution dipping. Surface roughness and homogeneity, elemental composition, and thickness of the monolayers were evaluated by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and ellipsometry. Molecular loadings for each monolayer on TiO(2) were quantified by quartz crystal microgravimetry (QCM), and the stability of bonding between each class of dyes and the TiO(2) was evaluated by measuring desorption, also by QCM; the carboxylates underwent significant dissociation in aqueous media but the phosphonates did not. DSSCs were prepared from each congener and from simple oligothiophene phosphonates to determine the effect of the cyanovinyl group on device behavior; all DSSCs were studied under irradiation from a AM 1.5G solar light source; the effect of cyanovinyl group termination was comparable to that of adding a thiophene moiety, and the DSSC using a self-assembled monolayer of (sexithiophene)phosphonate (6TP) had total power conversion efficiency (η) of ca. 5%.

Real-Time Fluorescence Turn-On Detection of Alkaline Phosphatase Activity with a Novel Perylene Probe

A tetracationic perylene probe (probe 1) was designed and synthesized. Probe 1 was used for the real-time fluorescence turn-on assay of alkaline phosphatase (ALP) activity and inhibitor screening. Probe 1 monomer fluorescence could be very efficiently quenched by ATP through the formation of an ATP/probe 1 complex. ALP triggered the degradation of ATP, the breakdown of the ATP/probe 1 complex, and the recovery of the probe 1 monomer fluorescence. In the presence of an ALP inhibitor, a decrease in fluorescence recovery was observed.

Tim-3-Expressing CD4(+) and CD8(+) T Cells in Human Tuberculosis (TB) Exhibit Polarized Effector Memory Phenotypes and Stronger Anti-TB Effector Functions

T-cell immune responses modulated by T-cell immunoglobulin and mucin domain-containing molecule 3 (Tim-3) during Mycobacterium tuberculosis (Mtb) infection in humans remain poorly understood. Here, we found that active TB patients exhibited increases in numbers of Tim-3-expressing CD4(+) and CD8(+) T cells, which preferentially displayed polarized effector memory phenotypes. Consistent with effector phenotypes, Tim-3(+)CD4(+) and Tim-3(+)CD8(+) T-cell subsets showed greater effector functions for producing Th1/Th22 cytokines and CTL effector molecules than Tim-3(-) counterparts, and Tim-3-expressing T cells more apparently limited intracellular Mtb replication in macrophages. The increased effector functions for Tim-3-expressing T cells consisted with cellular activation signaling as Tim-3(+)CD4(+) and Tim-3(+)CD8(+) T-cell subsets expressed much higher levels of phosphorylated signaling molecules p38, stat3, stat5, and Erk1/2 than Tim-3- controls. Mechanistic experiments showed that siRNA silencing of Tim-3 or soluble Tim-3 treatment interfering with membrane Tim-3-ligand interaction reduced de novo production of IFN-γ and TNF-α by Tim-3-expressing T cells. Furthermore, stimulation of Tim-3 signaling pathways by antibody cross-linking of membrane Tim-3 augmented effector function of IFN-γ production by CD4(+) and CD8(+) T cells, suggesting that Tim-3 signaling helped to drive stronger effector functions in active TB patients. This study therefore uncovered a previously unknown mechanism for T-cell immune responses regulated by Tim-3, and findings may have implications for potential immune intervention in TB.

UPR-Induced Resistance to Etoposide Is Downstream of PERK and Independent of Changes in Topoisomerase IIα Levels

The unfolded protein response (UPR) is regulated by three ER-localized, transmembrane signal transducers that control distinct aspects of the UPR. We previously reported that both increased resistance to etoposide and a reduction in Topoisomerase IIα protein levels were a direct response of UPR activation, and the latter occurred independent of changes in Topo IIα mRNA levels. We have now examined the contribution of each of the three up-stream transducers of the UPR, as well as some of their downstream targets in affecting decreased expression of Topo IIα protein and increased drug resistance.

Risk Factors of Pneumothorax After Endobronchial Ultrasound-guided Transbronchial Biopsy for Peripheral Lung Lesions

The risk of endobronchial ultrasound-guided transbronchial biopsy-related pneumothorax is a major concern and warrants further studies. The aim of our study was to estimate the risk of pneumothorax after this procedure and identify its risk factors.

Sensitive and Simultaneous Detection of Different Disease Markers Using Multiplexed Gold Nanorods

A multiplexed bioanalytical assay is produced by incorporating two types of gold nanorods (GNRs). Besides retaining the desirable features of common GNRs LSPR sensors, this sensor is easy to fabricate and requires only a visible-NIR spectrometer for detection. This assay can simultaneously detect different acceptor-ligand pairs by choosing the proper GNRs possessing various LPWs in a wide detection wavelength range and can be developed into a high-throughput detection method. This bioanalytical assay allows easy detection of human serum specimens infected by S. japonicum and tuberculosis (TB) from human serum specimens (human serum/Tris-HCl buffer ratio=1:10(4)) without the need for sample pretreatment. The technique is very sensitive compared to other standard methods such as indirect hemagglutination assays (IHA) that require a serum concentration ratio of larger than 1:20 and enzyme-linked immunosorbent assays (ELISA) requiring a ratio larger than 1:100. This methodology can be readily extended to other immunoassays to realize wider diagnostic applications.

Effects of Arsenite in Astrocytes on Neuronal Signaling Transduction

The main purpose of this study was to test the hypothesis that arsenite induces neurotoxicity via effects on astrocytes. Astrocytes were exposed to 0, 5 or 10μM arsenite in medium for 24h, and then astrocyte-conditioned medium (ACM) was collected after incubation with fresh medium for 6h. Primary neuron cultures were divided into four groups due to ACM, which were neurons without ACM exposure (group I) and neurons exposed to ACM from 0, 5 or 10μM arsenite treated astrocytes (group II-IV). Protein expression of N-methyl-d-aspartate receptors (NR1, NR2A, NR2B), calmodulin-dependent protein kinaseII (CaMKII) and adenylate cyclase (AC) in neurons were measured after incubation with ACM for 4, 8 or 12h. Morphological changes and synaptic formation were observed after a 72h-incubation with ACM. Compared to group II, synaptic formation and protein expression of NR2A, NR2B, CaMKII and AC in group III and IV were significantly suppressed. Moreover, synaptic formation and protein expression of CaMKII and AC in group II were significantly enhanced when compared with group I. Taken together, findings from this study suggested that arsenic in astrocytes might impair synaptic formation through disturbing astrocytic effects on neuronal signal transduction.

Comparison Between Magnitude Reconstruction and Phase-sensitive MR Imaging in the Detection of Myocardial Infarction

To determine the deference between phase sensitive magnetic resonance (MR) imaging and magnitude reconstruction to detect infracted myocardium.

Dendritic Cell-based Vaccination for Renal Cell Carcinoma: Challenges in Clinical Trials

After decades of research, dendritic cell (DC)-based vaccines for renal cell carcinoma have progressed from preclinical rodent models and safety assessments to Phase I/II clinical trials. DC vaccines represent a promising therapy that has produced measurable immunological responses and prolonged survival rates. However, there is still much room to improve in terms of therapeutic efficacy. The key issues that affect the efficiency and reliability of DC therapy include the selection of patients who will respond best to treatment, the proper preparation and administration of DC vaccines, and a combination of DC vaccination with other immune-enhancing therapies (e.g., removal of Tregs, CTLA-4 blockade and lymphodepletion). Additional antiangiogenic agents will hopefully lead to greater survival benefits for patients in early disease stages. This review focuses on the different approaches of DC-based vaccination against renal cell carcinoma and potential strategies to enhance the efficacy of DC vaccination.

WIN55, 212-2 Promotes Differentiation of Oligodendrocyte Precursor Cells and Improve Remyelination Through Regulation of the Phosphorylation Level of the ERK 1/2 Via Cannabinoid Receptor 1 After Stroke-induced Demyelination

In stroke, a common cause of neurological disability in adults is that the myelin sheaths are lost through the injury or death of mature oligodendrocytes, and the failure of remyelination may be often due to insufficient proliferation and differentiation of oligodendroglial progenitors. In the current study, we used middle cerebral artery occlusion (MCAO) to induced transient focal cerebral ischemia, and found that WIN55, 212-2 augmented actively proliferating oligodendrocytes measured by CC1 immunoreactive cells within the peri-infarct areas. To establish whether these effects were associated with changes in myelin formation, we analyzed the expression of myelin basic protein (MBP) and myelin ultrastructure. We found that WIN55, 212-2 showed more extensive remyelination than vehicle at 14 days post injection (dpi). The extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling pathway may be involved in OPCs differentiation. To determine the regulatory effect of WIN55, 212-2 post-treatment on phospho-ERK 1/2 (p-ERK 1/2) after ischemia/reperfusion, Western blot analysis was performed. We found that WIN55, 212-2 regulated the phosphorylation level of the ERK 1/2 to promote OPCs survival and differentiation. Notably, cannabinoid receptor 1 is coupled to the activation of the ERK cascade. Following rimonabant combined treatment, the effect of WIN55, 212-2 on regulating the phosphorylation level of the ERK 1/2 was reversed, and the effect of accelerated myelin formation was partially inhibited. Together, we first found that WIN55, 212-2 promoted OPCs differentiation and remyelination through regulation of the level of the p-ERK 1/2 via cannabinoid receptor 1.

Identification, Sequence Analysis and Characterization of Clonorchis Sinensis Ubiquitin

Ubiquitin is a functionally important protein expressed in eukaryotic cells usually encoded by multigenic families containing two types of genes, ubiquitin extension genes and polyubiquitin genes. One independent monomeric locus and two independent polyubiquitin loci were firstly identified from the genome of carcinogenic liver fluke, Clonorchis sinensis (C. sinensis). The nucleotide and amino acid sequence of C. sinensis polyubiquitin, especially polyubiquitin with five tandem ubiquitin repeats (CsPUB5), were analyzed. We obtained recombinant CsPUB5 (rCsPUB5) and anti-rCsPUB5 IgG. The ubiquitin transcripts in life cycle of C. sinensis were investigated. In addition, we found that ubiquitin or ubiquitination was ubiquitous in adult worm of C. sinensis and significantly observed in the content of biliary tract and intrahepatic biliary epithelium of liver from C. sinensis infected rat. We confirmed that rCsPUB5 could bind to human intrahepatic biliary epithelial cell by immunofluorescence in vitro. It was considered that ubiquitin family constitutively expressed in C. sinensis for variety of cellular processes and might be implicated in the genesis and progression of cholangiocarcinoma induced by the infection of C. sinensis.

Evaluating the Impact of Patient, Tumor, and Treatment Characteristics on the Development of Jaw Complications in Patients Treated for Oral Cancers: A SEER-Medicare Analysis

BACKGROUND: Jaw complications, including osteoradionecrosis, are significant sequelae of radiation therapy (RT) for oral cancers. This study identifies the impact of patient, tumor, and treatment characteristics on the development of jaw complications in patients treated with RT. METHODS: The Surveillance, Epidemiology, and End Results (SEER)-Medicare database was used to identify patients treated with RT for oral cancers from 1999 to 2007. Jaw complications were identified by International Classification of Diseases 9th revision (ICD-9) diagnosis codes and/or related procedures using Current Procedural Terminology (CPT) and ICD-9 codes. RESULTS: A total of 1848 patients were identified. With a median follow-up of 2.5 years, 297 patients (16.1%) developed jaw complications: 226 patients had a diagnosis, 41 patients had a procedure, and 30 patients had both. On multivariate analysis, female sex, lack of chemotherapy use, and fewer comorbidities were associated with a statistically significant increase in jaw complications. CONCLUSIONS: Even with modern techniques, jaw complications are a notable and potentially devastating side effect of RT for oral cancers. © 2012 Wiley Periodicals, Inc. Head Neck, 2012.

Scission of the P53-MDM2 Loop by Ribosomal Proteins

The oncoprotein MDM2 is both the transcriptional target and the predominant antagonist of the tumor suppressor p53. MDM2 inhibits the functions of p53 via a negative feedback loop that can be circumvented by several ribosomal proteins in response to nucleolar or ribosomal stress. Stress conditions in the nucleolus can be triggered by a variety of extracellular and intracellular insults that impair ribosomal biogenesis and function, such as chemicals, nutrient deprivation, DNA damaging agents, or genetic alterations. The past decade has witnessed a tremendous progress in understanding this previously underinvestigated ribosomal stress-MDM2-p53 pathway. Here, we review the recent progress in understanding this unique signaling pathway, discuss its biological and pathological significance, and share with readers our insight into the research in this field.

Rational Design of Highly Selective Spleen Tyrosine Kinase Inhibitors

A novel approach to design selective spleen tyrosine kinase (Syk) inhibitors is described. Inhibition of spleen tyrosine kinase has attracted much attention as a mechanism for the treatment of autoimmune diseases such as asthma, rheumatoid arthritis, and SLE. Fostamatinib, a Syk inhibitor that successfully completed phase II clinical trials, also exhibits some undesirable side effects. More selective Syk inhibitors could offer safer, alternative treatments. Through a systematic evaluation of the kinome, we identified Pro455 and Asn457 in the Syk ATP binding site as a rare combination among sequence aligned kinases and hypothesized that optimizing the interaction between them and a Syk inhibitor molecule would impart high selectivity for Syk over other kinases. We report the structure-guided identification of three series of selective spleen tyrosine kinase inhibitors that support our hypothesis and offer useful guidance to other researchers in the field.

Efficient 2-Aryl Benzothiazole Formation from Aryl Ketones and 2-Aminobenzenethiols Under Metal-Free Conditions

2-Aryl benzothiazole formation from aryl ketones and 2-aminobenzenethiols under metal- and I(2)-free conditions was described. Various 2-aryl benzothiazoles were selectively obtained in good yields using molecular oxygen as oxidant. DMSO played an important role in this transformation. Functional groups such as methyl, methoxy, fluoro, chloro, bromo and nitro groups were tolerated under the optimized reaction conditions.

Regarding: Comparison of Medial-to-Lateral Versus Traditional Lateral-to-Medial Laparoscopic Dissection Sequences for Resection of Rectosigmoid Cancers: Randomized Controlled Clinical Trial

Co-Synthesis of Cargo-Loaded Hydroxyapatite/Alginate Core-Shell Nanoparticles (HAP@Alg) As PH-Responsive Nanovehicles by a Pre-gel Method

A new core-shell nanostructure consisting of inorganic hydroxyapatite (HAP) nanoparticles as the core and organic alginate as the shell (denoted as HAP@Alg) was successfully synthesized by a pre-gel method and applied to pH-responsive drug delivery systems (DDS). HAP@Alg nanoparticles have the advantages of hydroxyapatite and alginate, where hydroxyapatite provides pH-responsive degradability, and alginate provides excellent biocompatibility and COOH functionality. Through the subsequent addition of CaCl2 and phosphate solutions to the alginate solution, HAP@Alg nanoparticles with controllable particle sizes (ranging from 160 to 650 nm) were obtained, and their core-shell structure was confirmed through transmission electron microscopy (TEM) observation. Rhodamine 6G (R6G), a positively charged dye, was selected as a model drug for pH-sensitive DDS. R6G was encapsulated in the HAP/Alg nanoparticles upon synthesis, and its loading efficiency could reach up to approximately 63.0%. The in vitro release behavior of the loaded R6G at different pH values was systematically studied, and the results indicated that more R6G molecules were released at lower pH conditions. For example, after releasing for 8 h, the release amount of R6G at pH 2.0 was 2.53-fold the amount at pH 7.4. We attributed this pH-sensitive release behavior to the dissolution of the HAP core in acidic conditions. The results of the MTT assay and confocal laser scanning microscopy indicated that the HAP@Alg were successfully uptaken by liver cancer cells (HepG2) without apparent cytotoxicity. The synthesized HAP@Alg nanoparticles show great potential as drug nanovehicles with high biocompatibility, enhanced drug loading, and pH-responsive features for future intracellular DDS.

Comparative Analysis of the Distribution of Segmented Filamentous Bacteria in Humans, Mice and Chickens

Segmented filamentous bacteria (SFB) are indigenous gut commensal bacteria. They are commonly detected in the gastrointestinal tracts of both vertebrates and invertebrates. Despite the significant role they have in the modulation of the development of host immune systems, little information exists regarding the presence of SFB in humans. The aim of this study was to investigate the distribution and diversity of SFB in humans and to determine their phylogenetic relationships with their hosts. Gut contents from 251 humans, 92 mice and 72 chickens were collected for bacterial genomic DNA extraction and subjected to SFB 16S rRNA-specific PCR detection. The results showed SFB colonization to be age-dependent in humans, with the majority of individuals colonized within the first 2 years of life, but this colonization disappeared by the age of 3 years. Results of 16S rRNA sequencing showed that multiple operational taxonomic units of SFB could exist in the same individuals. Cross-species comparison among human, mouse and chicken samples demonstrated that each host possessed an exclusive predominant SFB sequence. In summary, our results showed that SFB display host specificity, and SFB colonization, which occurs early in human life, declines in an age-dependent manner.The ISME Journal advance online publication, 15 November 2012; doi:10.1038/ismej.2012.128.

Identification and Characterization of a Gene Encoding a Putative Lysophosphatidyl Acyltransferase from Arachis Hypogaea

Adsorption Chromatography Separation of the Flavonols Kaempferol, Quercetin and Myricetin Using Cross-linked Collagen Fibre As the Stationary Phase

BACKGROUND: Kaempferol, quercetin and myricetin are typical flavonols that are most concentrated in many medicinal herbs. However, the separation of these flavonols is very challenging due to their similar molecular structures. In the present investigation, the chromatographic separation of the flavonols kaempferol, quercetin and myricetin was performed using glutaraldehyde cross-linked collagen fibre (GCF) as the stationary phase. RESULTS: Kaempferol, quercetin and myricetin could be completely separated from each other by the GCF column by using gradient elution with different solutions of aqueous ethanol (100% to 70%) and 50% acetone. When the chromatographic separation was carried out at a flow rate of 0.75 bed volume h(-1) with a sample loading of 30 mg 7 g(-1) GCF, the purity of kaempferol, quercetin and myricetin was 98.17%, 93.81% and 81.76%, respectively. The separation resolution was influenced by column length, flow rate and sample loading amount. The separation efficiency of GCF was not obviously reduced after applications had been repeated five times. In the fifth repeated application, the purity of the recovered kaempferol, quercetin and myricetin was still higher than 97%, 94% and 78%, respectively. CONCLUSION: GCF is a promising adsorbent for use as a stationary phase in the chromatographic separation of flavonols from their mixtures. © 2012 Society of Chemical Industry.

Enterobius Vermicularis Infection is Well Controlled Among Preschool Children in Nurseries of Taipei City, Taiwan

Whether Enterobius vermicularis (pinworm) infections among preschool children in Taipei City had truly declined was investigated.

Common FLG Mutation K4671X Not Associated with Atopic Dermatitis in Han Chinese in a Family Association Study

Filaggrin gene (FLG) mutations have been identified as the cause of ichthyosis vulgaris (IV) and major predisposing factors for atopic dermatitis (AD). The relationship among AD, IV and FLG mutations has not been clarified yet. Mutations 3321delA and K4671X, two of the most common mutations in Chinese patients, were both statistically associated with AD in case-control studies.

The Effect of Polymerized Placenta Hemoglobin on Renal Ischemia/reperfusion Injury

Abstract The goal of this study was to investigate whether hemoglobin-based oxygen carrier (HBOC) attenuated ischemia/reperfusion (I/R)-induced kidney injury. Male SD rats were randomly divided into a sham group, I/R group, and HBOC group (injection of 0.1 gHb/kg PolyPHb). The ischemia was induced by bilateral renal pedicle cross-clamping for 45min. Then the clamp was released to allow 24h reperfusion. Without increasing blood pressure, PolyPHb reduced the blood urea nitrogen and creatinine in plasma and attenuated the tumor necrosis factor-α and interleukin-8 in kidney tissue. Therefore, our findings suggest that PolyPHb could reduce kidney injury after I/R injury, and this effect was probably associated with the depressed inflammatory response.

Hepatitis C Testing, Infection, and Linkage to Care Among Racial and Ethnic Minorities in the United States, 2009-2010

Objectives. We estimated rates and determinants of hepatitis C virus (HCV) testing, infection, and linkage to care among US racial/ethnic minorities. Methods. We analyzed the Racial and Ethnic Approaches to Community Health Across the US Risk Factor Survey conducted in 2009-2010 (n = 53 896 minority adults). Results. Overall, 19% of respondents were tested for HCV. Only 60% of those reporting a risk factor were tested, with much lower rates among Asians reporting injection drug use (40%). Odds of HCV testing decreased with age and increased with higher education. Of those tested, 8.3% reported HCV infection. Respondents with income of $75 000 or more were less likely to report HCV infection than those with income less than $25 000. College-educated non-Hispanic Blacks and Asians had lower odds of HCV infection than those who did not finish high school. Of those infected, 44.4% were currently being followed by a physician, and 41.9% had taken HCV medications. Conclusions. HCV testing and linkage to care among racial/ethnic minorities are suboptimal, particularly among those reporting HCV risk factors. Socioeconomic factors were significant determinants of HCV testing, infection, and access to care. Future HCV testing and prevention activities should be directed toward racial/ethnic minorities, particularly those of low socioeconomic status. (Am J Public Health. Published online ahead of print November 15, 2012: e1-e8. doi:10.2105/AJPH.2012.300858).

Comprehensive Evaluation of the Adenovirus/alphavirus-replicon Chimeric Vector-based Vaccine RAdV-SFV-E2 Against Classical Swine Fever

Classical swine fever (CSF) is an economically important, highly contagious swine disease caused by classical swine fever virus (CSFV). Marker vaccines and companion serological diagnostic tests are thought to be a promising strategy for future control and eradication of CSF. Previously, we have demonstrated that an adenovirus-vectored Semliki forest virus replicon construct expressing the E2 glycoprotein from CSFV, rAdV-SFV-E2, induced sterile immunity against a lethal CSFV challenge. In this study, we further evaluated the vaccine with respect to its safety, number and dose of immunization, and effects of maternally derived antibodies, re-immunization of the vaccine or co-administration with pseudorabies vaccine on the vaccine efficacy. The results showed that: (1) the vaccine was safe for mice, rabbits and pigs; (2) two immunizations with a dose as low as 6.25×10(5)TCID(50) or a single immunization with a dose of 10(7)TCID(50) rAdV-SFV-E2 provided complete protection against a lethal CSFV challenge; (3) maternally derived antibodies had no inhibitory effects on the efficacy of the vaccine; (4) the vaccine did not induce interfering anti-vector immunity; and (5) co-administration of rAdV-SFV-E2 with a live pseudorabies vaccine induced antibodies and protection indistinguishable from immunization with either vaccine administered alone. Taken together, the chimeric vaccine represents a promising marker vaccine candidate for control and eradication of CSF.

Gallic Acid Inhibits Gastric Cancer Cells Metastasis and Invasive Growth Via Increased Expression of RhoB, Downregulation of AKT/small GTPase Signals and Inhibition of NF-κB Activity

Our previous study demonstrated the therapeutic potential of gallic acid (GA) for controlling tumor metastasis through its inhibitory effect on the motility of AGS cells. A noteworthy finding in our previous experiment was increased RhoB expression in GA-treated cells. The aim of this study was to evaluate the role of RhoB expression on the inhibitory effects of GA on AGS cells. By applying the transfection of RhoB siRNA into AGS cells and an animal model, we tested the effect of GA on inhibition of tumor growth and RhoB expression. The results confirmed that RhoB-siRNA transfection induced GA to inhibit AGS cells' invasive growth involving blocking the AKT/small GTPase signals pathway and inhibition of NF-κB activity. Finally, we evaluated the effect of GA on AGS cell metastasis by colonization of tumor cells in nude mice. It showed GA inhibited tumor cells growth via the expression of RhoB. These data support the inhibitory effect of GA which was shown to inhibit gastric cancer cell metastasis and invasive growth via increased expression of RhoB, downregulation of AKT/small GTPase signals and inhibition of NF-κB activity. Thus, GA might be a potential agent in treating gastric cancer.

Carbamazepine-induced Hypersensitivity Syndrome in Chronic Schizophrenia

Drug-induced hypersensitivity syndrome is a clinically important issue. We report a case of carbamazepine-induced hypersensitivity syndrome in a 35-year-old schizophrenia patient. This patient had no previous food or medication allergy history and presented a negative test result of HLA-B*1502 genotype. After 19 days exposure of carbamazepine, high fever up to 39.4°C, leucopenia (1670/mm(3)), proteinuria and bilateral lung field infiltration were developed. These clinically significant physical conditions resolved after discontinuing carbamazepine. The importance of genetic susceptibility other than HLA-B*1502 should not be overlooked in drug-induced hypersensitivity syndrome.

Functional Magnetic Resonance Imaging of Methamphetamine Craving

The study aimed to explore the abnormal activation of special brain areas associated with methamphetamine craving using functional magnetic resonance imaging (fMRI) and to reveal the neurobiological basis of addiction. Twenty-six methamphetamine addicts and 26 healthy subjects were scanned by brain fMRI while watching pictures of happy, sad, or methamphetamine to acquire resource data. SPM5 was used to analyze fMRI data to get related brain activation map, and it was found that methamphetamine addicts had high brain activation in cingulate and low activation in frontal lobe when watching methamphetamine-cue pictures. This study demonstrated that methamphetamine addicts had emotion-related brain activation abnormalities.

A Comparison of Pyelography and Various Reconstructions of Multidetector Helical Computed Tomography Urography Images for Diagnosing Urinary Obstruction

Radiologists and urologists require practical and helpful image reconstructions for diagnosing urinary obstruction. We performed different types of imaging and reconstruction, then used a self-designed urinary obstruction-specific questionnaire to evaluate the diagnostic outcome of them. Our results suggested that two-dimensional (2D) axial computed tomography (CT) is clinically superior to retrograde pyelography or antegrade pyelography, and to other modes of image reconstruction that are often used for diagnosing urinary obstruction.

ATM Polymorphisms Predict Severe Radiation Pneumonitis in Patients With Non-Small Cell Lung Cancer Treated With Definitive Radiation Therapy

PURPOSE: The ataxia telangiectasia mutated (ATM) gene mediates detection and repair of DNA damage. We investigated associations between ATM polymorphisms and severe radiation-induced pneumonitis (RP). METHODS AND MATERIALS: We genotyped 3 potentially functional single nucleotide polymorphisms (SNPs) of ATM (rs1801516 [D1853N/5557G>A], rs189037 [-111G>A] and rs228590) in 362 patients with non-small cell lung cancer (NSCLC), who received definitive (chemo)radiation therapy. The cumulative severe RP probabilities by genotypes were evaluated using the Kaplan-Meier analysis. The associations between severe RP risk and genotypes were assessed by both logistic regression analysis and Cox proportional hazard model with time to event considered. RESULTS: Of 362 patients (72.4% of non-Hispanic whites), 56 (15.5%) experienced grade ≥3 RP. Patients carrying ATM rs189037 AG/GG or rs228590 TT/CT genotypes or rs189037G/rs228590T/rs1801516G (G-T-G) haplotype had a lower risk of severe RP (rs189037: GG/AG vs AA, adjusted hazard ratio [HR] = 0.49, 95% confidence interval [CI], 0.29-0.83, P=.009; rs228590: TT/CT vs CC, HR=0.57, 95% CI, 0.33-0.97, P=.036; haplotype: G-T-G vs A-C-G, HR=0.52, 95% CI, 0.35-0.79, P=.002). Such positive findings remained in non-Hispanic whites. CONCLUSIONS: ATM polymorphisms may serve as biomarkers for susceptibility to severe RP in non-Hispanic whites. Large prospective studies are required to confirm our findings.

Distinct Requirements for Wnt9a and Irf6 in Extension and Integration Mechanisms During Zebrafish Palate Morphogenesis

Development of the palate in vertebrates involves cranial neural crest migration, convergence of facial prominences and extension of the cartilaginous framework. Dysregulation of palatogenesis results in orofacial clefts, which represent the most common structural birth defects. Detailed analysis of zebrafish palatogenesis revealed distinct mechanisms of palatal morphogenesis: extension, proliferation and integration. We show that wnt9a is required for palatal extension, wherein the chondrocytes form a proliferative front, undergo morphological change and intercalate to form the ethmoid plate. Meanwhile, irf6 is required specifically for integration of facial prominences along a V-shaped seam. This work presents a mechanistic analysis of palate morphogenesis in a clinically relevant context.

Association and Prognostic Value of Serum Inflammation Markers in Patients with Leukoplakia and Oral Cavity Cancer

Abstract Background: Oral cavity cancer ranks as the fourth leading cancer in men in Taiwan. The development of a serum biomarker panel for early detection and disease monitoring is, therefore, warranted. Methods: Nine inflammation-associated markers were investigated in 46 patients with leukoplakia, 151 patients with untreated oral cavity squamous cell carcinoma (OSCC), and 111 age- and gender-matched healthy controls using enzyme-linked immunosorbent assay. During a subsequent 28-month surveillance of OSCC patients, serum samples were prospectively collected at predetermined intervals following the completion of therapy. Results: Logistic regression analysis showed matrix metalloproteases (MMP)-2, MMP-9, C-reactive protein (CRP), transforming growth factor-β1 (TGF-β1), and E-selectin having the best discrimination power between groups and significant elevation trends of those five markers were noted from control to OSCC. By combining those five markers, a 0.888 and 0.938 area under curve by ROC curve analysis with 67.4% and 80% overall sensitivity and fixed 90% specificity for leukoplakia and OSCC groups were demonstrated. In the follow-up period, 25 OSCC patients developed recurring or secondary tumors. All examined markers had decreased in relapse-free patients following treatment. However, in patients with relapse, interleukin-6, CRP, and serum amyloid A remained at elevated levels. Statistical analysis showed that patients with CRP ≧2 mg/L and E-selectin ≧85 ng/mL at baseline had highest probability of relapse (odds ratio=3.029, p<0.05). Conclusions: The results indicate that inflammation plays a crucial role in the pathogenesis process of OSCC. By examining the inflammation markers, physicians could potentially identify patients at risk of cancer transformation or relapse.

Bacterial Colony from Two-dimensional Division to Three-dimensional Development

On agar surface, bacterial daughter cells form a 4-cell array after the first two rounds of division, and this phenomenon has been previously attributed to a balancing of interactions among the daughter bacteria and the underneath agar. We studied further the organization and development of colony after additional generations. By confocal laser scanning microscopy and real-time imaging, we observed that bacterial cells were able to self-organize and resulted in a near circular micro-colony consisting of monolayer cells. After continuous dividing, bacteria transited from two-dimensional expansion into three-dimensional growth and formed two to multi-layers in the center but retained a monolayer in the outer ring of the circular colony. The transverse width of this outer ring appeared to be approximately constant once the micro-colony reached a certain age. This observation supports the notion that balanced interplays of the forces involved lead to a gross morphology as the bacteria divide into offspring on agar surface. In this case, the result is due to a balance between the expansion force of the dividing bacteria, the non-covalent force among bacterial offspring and that between bacteria and substratum.

Suppression of Tongue Squamous Cell Carcinoma Growth by Inhibition of Jagged1 in Vitro and in Vivo

BACKGROUND: The changes in Notch signaling are closely related to the occurrence and development of many cancers. We have investigated Notch signaling receptor and its ligand expressions in TSCC cell lines, tissues and its significance. We clarified Notch signaling pathway in TSCC and its mechanism. We regulated Notch signaling pathway of tumor cells, thereby inhibiting tumor cell proliferation and differentiation. METHODS: We detected Jagged1 protein and mRNA expression levels in specimens (tongue cancer and adjacent tissues) from 74 patients with tongue cancer and in TSCC cell line. The Jagged1-targeted lentiviral vector RNAi system was constructed, and its suppressive effects on the proliferation and invasion of tongue carcinoma cells in in vivo and ex vivo were determined. RESULTS: Jagged1 was expressed in tongue squamous cell cancer tissues and cell line, but there were differences in its expression. Jagged1 was knocked down and the tumor growth was inhibited accompanying cell cycle changes. Animal studies also showed that the tumor growth was inhibited. CONCLUSIONS: Jagged1 may be involved in the differentiation and proliferation of tongue cancer. Targeting Jagged1 RNA interference lentiviral vector can effectively lower Jagged1 mRNA and protein expression levels of Tca8113 cells, thereby preventing the proliferation of TSCC cells. Jagged1 is expected to be a promising new target for curing tongue cancer. In-depth study of the interaction between Jagged1 and other molecules of Notch signaling pathway in the process of carcinogenesis has important theoretical guidance and clinical significance in revealing the mechanism of Jagged1 and its application in the therapy for tongue cancer.

Association of Maternal Education with the Neuroblastoma Susceptibility in Children: A Meta-Analysis

Maternal education might be an important factor for the neuroblastoma risk in children, but it was conflicting. This meta-analysis was performed to evaluate the relationship between maternal education and neuroblastoma susceptibility and to explore whether maternal education was an important indicator to be associated with the neuroblastoma risk in children. The association studies were identified from the databases of PubMed, and Cochrane Library as of June 1, 2012, and eligible investigations were synthesized using meta-analysis method. Results were expressed with odds ratios (OR) for dichotomous data, and 95% confidence intervals (CI) were also calculated. Six literatures were identified for the analysis of association between maternal education and neuroblastoma susceptibility in children, consisting of 2063 patients with cancer and 13,925 controls. There was no a marked association between maternal education and neuroblastoma susceptibility when the maternal education was less than high school (OR = 0.66, 95% CI: 0.43-1.01, P = .06). We also found that maternal education was not associated with the neuroblastoma susceptibility when the maternal education was high school (OR = 0.74, 95% CI: 0.31-1.75, P = .49) and more than high school (OR = 0.78, 95% CI: 0.33-1.85, P = .58). In conclusion, maternal education is not associated with the neuroblastoma susceptibility in children. However, more investigations are required to further clarify the association of maternal education with the neuroblastoma susceptibility in children.

Ultralow-Threshold Two-Photon Pumped Amplified Spontaneous Emission and Lasing from Seeded CdSe/CdS Nanorod Heterostructures

Ultralow-threshold two-photon pumped amplified spontaneous emission (2ASE) and lasing in seeded CdSe/CdS nanodot/nanorod heterostructures is demonstrated for the first time. Such heterostructures allow the independent tunability of the two-photon absorption (2PA) cross-section (σ(2)) through varying the CdS rod size, and that of the emission wavelength through varying the CdSe dot size. With an enhanced σ(2), 2ASE in these heterostructures is achieved with an ultralow threshold fluence of ∼1.5 mJ/cm(2), which is as much as one order less than that required for spherical semiconductor NCs. Importantly, by exploiting this unique property of the seeded nanorods exhibiting strong quantum confinement even at relatively large rod sizes, a near reciprocal relation between the 2ASE threshold and the 2PA action cross-section (σ(2)η) (where η is the quantum yield) was found and validated over a wide volume range for II-VI semiconductor nanostructures. Ultrafast optical spectroscopy verified that while the Auger processes in these heterostructures are indeed suppressed, ASE in these samples could also be strongly affected by a fast hole-trapping process to the NR surface states. Lastly, to exemplify the potential of these seeded CdSe/CdS nanodot/nanorod heterostructures as a viable gain media for achieving two-photon lasing, a highly photostable microsphere laser with an ultralow pump threshold is showcased.

[Thallium Poisoning: Report of an Autopsy Case]

[The Development on Surveying and Monitoring Physical Fitness and Health Among Chinese Students and the Establishment of Its System]

[Complete Video-assisted Thoracoscopic Anatomic Segmentectomy for Pulmonary Diseases: the Early Experiences]

To evaluate the safety and feasibility of video-assisted thoracic surgery (VATS) anatomic segmentectomy for pulmonary diseases.

Changes in Expression of Nogo Receptor 1 in Hippocampus and Cortex After Cardiopulmonary Resuscitation in Rats

The aim of this study was to investigate changes in Nogo receptor 1 (NgR(1)) expression in the cerebrum after cardiopulmonary resuscitation (CPR) in rats. Cardiac arrest was induced by alternating current in 50 SD rats through transcutaneous electrical epicardium stimulation, and CPR was performed with the Utstein mode 6 minutes after cardiac arrest. Rats were killed 1, 3, and 7 days after CPR. We performed immunofluorescence with antibodies against NgR(1) to map the distribution of NgR(1) in the rat cerebrum, whereas quantitative polymerase chain reaction was performed for quantitative analysis of NgR(1) messenger RNA (mRNA). There was a striking transient up-regulation of the NgR(1) protein and mRNA in both the hippocampus and cortex in response to CPR. Nogo receptor 1 proteins were strongly expressed in hippocampal neurons 1 and 3 days after CPR (P < .001 for 1 day and P < .05 for 3 days, vs the control group, respectively), which returned to the basal level 7 days after CPR. In the cortex, staining moderately increased 1 day after CPR and got the peak level after 3 days (P < .001), returning to normal expression levels on day 7. The levels of NgR(1) mRNA in the hippocampus and cerebral cortical cortex showed the same trend with staining. The changes were significantly different between day 3 and baseline in both the hippocampus and cortex (P < .05, respectively). Furthermore, there were significant differences between the hippocampus and cerebral cortical cortex at 1 day and 3 days after the CPR (P < .05, respectively). There was a transient increase in NgR(1) in the vulnerable areas of the rat brain after CPR. Blockade of NgR(1) may be important in maintaining the high regenerative capacity of neurons during the time window when NgR(1) expression increases.

Determinant Factors of Gender Identity: A Commentary

Paediatric specialists involved in the care of children with disorders of sex development may be expected to provide straightforward answers to questions concerning the "true sex" of a child, reflecting common perceptions of sex/gender as an immutable binary biological reality. This article highlights how much more broad and complex the topic of gender identity and its development is. Many theories have been put forward to advance knowledge of gender identity. Against the breadth and depth of this vast topic, the current overview is inevitably incomplete. It begins by arguing for a more consistent use of 'sex' and 'gender'. It considers in turn three influential theoretical frameworks that lend themselves to empirical research. These are: 1) the role of the brain; 2) the role of socialisation; and 3) multi-dimensional gender development. The article ends by suggesting potentially fruitful questions and areas for future research.

[The Effects of Insulin and Gliclazide Therapy on Endoplasmic Reticulum Stress and Insulin Sensitivity in Liver of Type 2 Diabetic Rats]

To investigate the effect of insulin and gliclazide therapy on endoplasmic reticulum (ER) stress and insulin sensitivity in the liver of type 2 diabetic rats.

Value of Serum Procalcitonin Levels in Predicting Spontaneous Bacterial Peritonitis

Background/Aims: Spontaneous bacterial peritonitis (SBP) is a life-threatening disease that poses a great diagnostic challenge to clinicians. We aimed to systemically and quantitatively summarize the current evidence on the diagnostic value of the procalcitonin (PCT) test in identifying SBP. Methodology: We searched EMBASE, MEDLINE, the Cochrane database and reference lists of relevant articles with no language restrictions through May 2012. We selected original research that reported the diagnostic performance of PCT alone or compared with other biomarkers to diagnose SBP. We summarized test performance characteristics using forest plots, summary receiver operating characteristic curves and bivariate random effects models. Results: We found only three qualifying studies examining 181 episodes of suspected infection with 50 (27.6%) confirmed SBP episodes from 3 countries. Bivariate pooled sensitivity, specificity, positive likelihood ratios and negative likelihood ratios were 86% (95%CI: 73%-94%), 80% (95%CI: 72%-87%), 7.73 (95%CI: 0.91-65.64) and 0.14 (95%CI: 0.01-1.89), respectively. The global measures of accuracy, area under the receiver operating curve (AUROC) and diagnostic odds ratio (DOR), showed PCT has excellent discriminative capability and individual study showed serum PCT testing has better accuracy than ascitic PCT, serum CRP or IL-6 testing. There was evidence of significant heterogeneity but no evidence of publication bias. Conclusions: The existing literature suggests moderate to high accuracy for PCT as a diagnostic aid for SBP. However, larger, appropriately designed prospective studies are needed to conclusively address the value of serum PCT testing in SBP diagnosis.

Atrioventricular Node Reentrant Tachycardia in Patients with Congenitally Corrected Transposition of the Great Arteries and Results of Radiofrequency Catheter Ablation

BACKGROUND: -We sought to investigate the feasibility of radiofrequency catheter ablation (RFCA) of atrioventricular node reentrant tachycardia (AVNRT) and ideal site for slow pathway (SP) ablation in Congenitally Corrected Transposition of the Great Arteries (CCTGA). METHODS AND RESULTS: -Nine patients with CCTGA referred for catheter ablation of AVNRT were studied. A single His potential was recorded in 8 patients (89%, 6 {S,L,L}and 2 {I,D,D}). The earliest atrial activation during retrograde atrioventricular node (AVN) conduction occurred at His bundle region (HBE) (n=7) or shifting from HBE to coronary sinus ostium (n=1, {S,L,L}). Two anatomically separate His potentials were recorded in 1 patient (11%, {S,L,L}), one at the anteroseptum (HBE-1), the other at the confluence of the pulmonary and mitral annulus(HBE-2). In 8 cases with a single His potential recorded, SP was abated at the posterior-mid septum: 2 ({S,L,L}) at the right posteroseptum, 1 ({S,L,L}) at the left posteroseptum, and 5 (3{S,L,L} and 2{I,D,D}) at the midseptum after failure of energy application at the posteroseptum. Junctional rhythm was observed during RFCA in all 8 cases. In the remaining patient with two anatomically separate His potentials recorded, SP was successfully ablated from the confluence of the pulmonary and mitral annulus, slightly below the HBE-2. Junctional rhythm was also induced during RFCA. CONCLUSIONS: -In {S,L,L} or {I,D,D}, RFCA of AVNRT is feasible. SP input region can mainly be found in the posterior-mid septum, especially in patients with single penetrating AVNs. SP could usually be successfully ablated in these regions.

Recent Advances in Chitosan-based Nanoparticles for Oral Delivery of Macromolecules

Chitosan (CS), a cationic polysaccharide, is widely regarded as a safe and efficient intestinal absorption enhancer of therapeutic macromolecules, owing to its inherent mucoadhesive feature and ability to modulate the integrity of epithelial tight junctions reversibly. By using CS-based nanoparticles, many studies have attempted to protect the loaded macromolecules against acidic denaturation and enzymatic degradation, prolong their intestinal residence time, and increase their absorption by the intestinal epithelium. Derivatives of CS such as quaternized CS, thiolated CS and carboxylated CS have also been examined to further enhance its effectiveness in oral absorption of macromolecular drugs. This review article describes the synthesis of these CS derivatives and their characteristics, as well as their potential transport mechanisms of macromolecular therapeutics across the intestinal biological membrane. Recent advances in using CS and its derivatives as carriers for oral delivery of hydrophilic macromolecules and their effects on drug transport are also reviewed.

HMG-CoA Reductase Inhibitors Activate Caspase-1 in Human Monocytes Depending on ATP Release and P2X7 Activation

Recent studies have demonstrated the stimulatory effects of HMG-CoA reductase inhibitors, statins, on IL-1β secretion in monocytes and suggest a crucial role for isoprenoids in the inhibition of caspase-1 activity. In this study, we further elucidated the molecular mechanisms underlying the stimulatory effects of statins on caspase-1. Three commonly recognized mechanistic models for NLRP3 inflammasome activation (i.e., ATP/P2X7/K(+) efflux, ROS production, and lysosomal rupture) were investigated in statin-stimulated human THP-1 monocytes. We found that fluvastatin and lovastatin can synergize with LPS to trigger inflammasome activation. Moreover, statin-induced caspase-1 activation and IL-1β production in LPS-primed THP-1 cells are dependent on GGPP deficiency and P2X7 activation. In particular, increased ATP release accounts for the action of statins in P2X7 activation. We also provide evidence that statin-induced moderate ROS elevation is involved in this event. Moreover, the cathepsin B inhibitor was shown to reduce statin-induced IL-1β secretion. Consistently statins can induce cathepsin B activation and lysosomal rupture, as evidenced by LysoTracker staining. Statins also increase intracellular ATP secretion and IL-1β release in primary human monocytes and murine macrophages. Notably, exogenous ATP-elicited P2X7 activation and consequent IL-1β release, an index of direct NLRP3 inflammasome activation, were not altered by statins. Taken together, statin-induced enhancement of inflammasome activation in monocytes and macrophages covers multiple mechanisms, including increases in ATP release, ROS production, and lysosomal rupture. These data not only shed new insight into isoprenylation-dependent regulation of caspase-1 but also unmask mechanisms for statin-elicited inflammasome activation.

Endothelial Kruppel-like Factor 4 Protects Against Atherothrombosis in Mice

The endothelium regulates vascular homeostasis, and endothelial dysfunction is a proximate event in the pathogenesis of atherothrombosis. Stimulation of the endothelium with proinflammatory cytokines or exposure to hemodynamic-induced disturbed flow leads to a proadhesive and prothrombotic phenotype that promotes atherothrombosis. In contrast, exposure to arterial laminar flow induces a gene program that confers a largely antiadhesive, antithrombotic effect. The molecular basis for this differential effect on endothelial function remains poorly understood. While recent insights implicate Kruppel-like factors (KLFs) as important regulators of vascular homeostasis, the in vivo role of these factors in endothelial biology remains unproven. Here, we show that endothelial KLF4 is an essential determinant of atherogenesis and thrombosis. Using in vivo EC-specific KLF4 overexpression and knockdown murine models, we found that KLF4 induced an antiadhesive, antithrombotic state. Mechanistically, we demonstrated that KLF4 differentially regulated pertinent endothelial targets via competition for the coactivator p300. These observations provide cogent evidence implicating endothelial KLFs as essential in vivo regulators of vascular function in the adult animal.

Steinstrasse Formation After Extracorporeal Shock Wave Lithotripsy for Pancreatic Stones

Exome Sequencing-driven Discovery of Coding Polymorphisms Associated with Common Metabolic Phenotypes

AIMS/HYPOTHESIS: Human complex metabolic traits are in part regulated by genetic determinants. Here we applied exome sequencing to identify novel associations of coding polymorphisms at minor allele frequencies (MAFs) >1% with common metabolic phenotypes. METHODS: The study comprised three stages. We performed medium-depth (8×) whole exome sequencing in 1,000 cases with type 2 diabetes, BMI >27.5 kg/m(2) and hypertension and in 1,000 controls (stage 1). We selected 16,192 polymorphisms nominally associated (p < 0.05) with case-control status, from four selected annotation categories or from loci reported to associate with metabolic traits. These variants were genotyped in 15,989 Danes to search for association with 12 metabolic phenotypes (stage 2). In stage 3, polymorphisms showing potential associations were genotyped in a further 63,896 Europeans. RESULTS: Exome sequencing identified 70,182 polymorphisms with MAF >1%. In stage 2 we identified 51 potential associations with one or more of eight metabolic phenotypes covered by 45 unique polymorphisms. In meta-analyses of stage 2 and stage 3 results, we demonstrated robust associations for coding polymorphisms in CD300LG (fasting HDL-cholesterol: MAF 3.5%, p = 8.5 × 10(-14)), COBLL1 (type 2 diabetes: MAF 12.5%, OR 0.88, p = 1.2 × 10(-11)) and MACF1 (type 2 diabetes: MAF 23.4%, OR 1.10, p = 8.2 × 10(-10)). CONCLUSIONS/INTERPRETATION: We applied exome sequencing as a basis for finding genetic determinants of metabolic traits and show the existence of low-frequency and common coding polymorphisms with impact on common metabolic traits. Based on our study, coding polymorphisms with MAF above 1% do not seem to have particularly high effect sizes on the measured metabolic traits.

Functional Characterization of Three Mouse Formyl Peptide Receptors

The evolutionary relationship and functional correlation between human formyl peptide receptors (FPRs) and their mouse counterparts remain incompletely understood. We examined 3 members of the mouse formyl peptide receptor subfamily (mFprs) and found that they differ in agonist preference and cellular distributions. When stably expressed in transfected RBL-2H3 cells, mFpr1 was readily activated by N-formylated peptides derived from Listeria monocytogenes (fMIVTLF), Staphylococcus aureus (fMIFL) and mitochondria (fMMYALF). In contrast, the Escherichia coli-derived fMLF was 1,000-fold less potent. The above peptides were much less efficacious at mFpr2, which responded better to the synthetic hexapeptide WKYMVm, the synthetic agonists Quin-C1 (a substituted quinazolinone), and Compound 43 (a nitrosylated pyrazolone derivative). Saturation binding assays showed that mFpr1 and mFpr2 were expressed at similar levels on the cell surface, although their affinity for fMLFIIK-FITC varied by more than 1,000-fold (K(d) values of 2.8 nM for mFpr1 and 4.8 μM for mFpr2). Contrary to these receptors, mFpr-rs1 responded poorly to all peptides tested above. Fluorescent microscopy revealed an intracellular distribution pattern of mFpr-rs1. Based on these results, we conclude that mFpr1 is an orthologue of human FPR1 with certain pharmacological properties of human FPR2/ALX, whereas mFpr2 has much lower affinity for formyl peptides. The intracellular distribution of mFpr-rs1 suggests an evolutionary correlation with human FPR3.

BOLD Responses to Different Temporospatial Frequency Stimuli in V1 and V2 Visual Cortex of Anisometropic Amblyopia

Purpose. Functional magnetic resonance imaging (fMRI) is the most advanced neuroimaging technique. The aim of this study was to investigate the blood oxygenation level-dependent (BOLD) of V1 and V2 visual cortex in anisometropic amblyopia with fMRI and explore the neural mechanism of amblyopia.
Methods. fMRI was performed with a 3.0-T MRI scanner during reversal checkerboard visual stimulation with different spatial frequencies (SF) of 0.4, 2, and 8 cpd in 2 states of temporal frequencies (TF) of 6 Hz and 8 Hz in a group of patients with anisometropic amblyopia (n=5) and a group of normal observers (n=4). Data were processed by SPM software offline. Responses of different eyes were compared in different conditions.
Results. The BOLD signal magnitude in V1 and V2 visual cortex of amblyopic eyes was significantly lower than the fellow eyes with anisometropic amblyopia at low SF (0.4-2 cpd) (p<0.05), but it was significantly higher than the fellow eyes at high SF (8 cpd) (p<0.05). The BOLD signal magnitude in V1 and V2 visual cortex of amblyopic eyes was significantly lower than the nondominant eyes in normal subjects in all conditions (p<0.001).
Conclusions. There are cortical deficits in V1 and V2 visual cortex of anisometropic amblyopia, which may be useful for selecting an optimum stimulus at proper temporospatial frequency.

Age-related Changes in Body Composition and Their Relationship with Bone Mineral Density Decreasing Rates in Central South Chinese Postmenopausal Women

The purpose of this work is to investigate the age-related changes in body composition and their relationship with bone mineral density decreasing rates (BDR) in central south Chinese postmenopausal women. BDR is the percentage of bone mineral density (BMD) decreasing value relative to the peak bone mass. A cross-sectional study was conducted on 779 healthy postmenopausal women, aged 50-77. Lumbar spine, total hip, and femoral neck BMD and body composition were measured by dual-energy X-ray absorptiometry. In women under 65, lean mass levels showed a stable downward trend, and were significantly higher than those of the 65-70 and >70 age groups; however, the fat mass levels showed no significant difference between the age groups. After controlling for age, age at menopause, and height, both fat mass and lean mass positively correlated with BDR at the lumbar(1-4) spine, the femoral neck and the total hip. When BDR at the lumbar(1-4) spine was used as the dependent variable, a higher R (2) change and partial R (2) were seen in fat mass than the age, age at menopause or lean mass, indicating that fat mass was the most significant determinant of BDR at this site. When BDR at the femoral neck or total hip was used as the dependent variable, respectively, lean mass was a more significant determinant than that of fat mass. We found that with advancing age, lean mass begins to decrease in women aged over 65 years, but fat mass levels show no significant difference between the age groups. Both fat mass and lean mass positively correlate with BDR, with site-specific differences. Fat mass is the most significant determinant of BDR at the lumbar spine, whereas lean mass is the most significant determinant of BDR at the femoral neck and total hip.

Ultrasound Microbubble Contrast Agent-mediated Suicide Gene Transfection in the Treatment of Hepatic Cancer

The aim of this study was to investigate the role of ultrasound microbubble contrast agent-mediated suicide gene transfection in the treatment of hepatic cancer. We intratumorally injected KDR-TK, AFP-TK and microbubble contrast agent into nude mice prior to ultrasound treatment and administration of prodrugs (GCV and 5-FC). The tumor volume, tumor inhibition rate, survival time and apoptosis of tumor cells was determined. The sizes of subcutaneous hepatic cancers in mice receiving treatment were comparable to those in the control group, and the survival time was similar between the two groups (P>0.05). However, the tumor inhibition rate and the number of apoptotic cells in the treatment group was markedly higher compared with that in the control group (P<0.05). Evident tumor necrosis was absent in both groups, except at the needle tract. Ultrasound therapy following injection of suicide genes and microbubble contrast agents is able to inhibit cancer growth in vivo. This may be attributed to the induction of cancer cell apoptosis.

Letter: Statin Use and the Risk of Oesophageal Cancer

[Clinical Role of F-18 FDG PET/CT in Differentiating Malignant and Benign Pleural Effusion in Patients with Lung Cancer]

Pleural effusions, a common feature among patients with lung cancer, should be differentiated into benign or malignant pleural effusions. F-18 fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) is one of accurate diagnostic tool for differentiating benign from malignant disease and for mediastinal lymph node staging. We explored the clinical role of PET/CT for differentiating malignant pleural effusion from benign pleural effusion among patients with lung cancer.

Association of IL12B Polymorphisms with Susceptibility to Graves Ophthalmopathy in a Taiwan Chinese Population

ABSTRACT:

Embedded Coupler Based on Selectively Infiltrated Photonic Crystal Fiber for Strain Measurement

A photonic crystal fiber (PCF) with embedded coupler is demonstrated for strain measurement. The embedded coupler is constructed by the selective filling of one of the air holes in the PCF. Light propagated in the fiber core can be efficiently coupled to the liquid-filled rod waveguide under phase-matching conditions, resulting in sharp decreasing of resonant wavelength intensity. The highest strain sensitivity is calculated to be ∼23.8  pm/με due to the coupling between core mode and fundamental mode of the liquid rod, when the refractive index (RI) of the liquid is 1.46. With the increase of the RI, the resonance can also be observed between the core mode and the higher-order modes of the liquid rod, whereas the strain sensitivity drops to ∼6.4  pm/με. The experimentally obtained static strain sensitivity values are ∼22 and ∼3.8  pm/με for the coupling between the core mode and the fundamental mode or linearly polarized LP11 modes of the liquid rod, respectively, which are in good agreement with the simulations. The dynamic strain measurement resolution obtained is ∼1.2  nε/(Hz)1/2.

Effect of HIV on Survival in Patients with Non-small-cell Lung Cancer in the Era of Highly Active Antiretroviral Therapy: a Population-based Study

HIV-infected patients with lung cancer have been reported to have poorer survival than uninfected patients. Whether this outcome holds true in the era of highly active antiretroviral therapy (HAART) is unclear. We examined the effect of HIV infection on clinical outcome in patients with lung cancer who are also receiving HAART.

Stabilization of Zeylenone in Rat Plasma by the Presence of Esterase Inhibitors and Its LC-MS/MS Assay for Pharmacokinetic Study

Six esterase inhibitors, namely EDTA·2Na(+) , NaF, phenylmethanesulfonyl fluoride, dichlorvos, bis-nitrophenyl phosphate (BNPP) and thenoyltrifluoroacetone, and the mixture of NaF and BNPP, were evaluated for the stabilization of labile benzoate containing zeylenone in rat plasma. The mixture appeared to exhibit the most effectively stabilizing effect with the degraded content of zeylenone decreasing from >60% (in the absence of inhibitors) to <6%. Following the stabilization by the addition of NaF (5 m m) and BNPP (5 m m), the analytes in rat plasma were acidified by formic acid and extracted into ethyl acetate at 0°C. After chromatographic separation, the detection of zeylenone was performed on a 3200 Q-Trap with positive ion electrospray mode, monitoring the ion transition m/z 383.2 → 105.0. The method was validated over the range from 2.68 to 1340 ng/mL with inter- and intra-run precision for the quality control samples being less than 6.8%. The assay accuracy was within 100 ± 7.0%. The validated method was successfully applied to a pharmacokinetic study in rats after the intratracheal administration of zeylenone in free drug or polymeric micellar solutions. The results showed that the pulmonary absorption of zeylenone loaded in micelles was significantly retarded compared with that of free drug solutions. Copyright © 2012 John Wiley & Sons, Ltd.

Cell Viability and Angiogenic Potential of a Bioartificial Adipose Substitute

An implantable scaffold pre-seeded with cells needs to remain viable and encourage rapid angiogenesis in order to replace injured tissues, especially for tissue defect repairs. We created a bioartificial adipose graft composed of an electrospun 3D nanofibrous scaffold and fat tissue excised from New Zealand white rabbits. Cell viability and angiogenesis potential of the bioartificial substitute were examined during four weeks of culture in Dulbecco's Modified Eagle Medium by immunohistochemical staining with LIVE/DEAD® cell kit and PECAM-1 antibody, respectively. In addition, a Matrigel® assay was performed to examine the possibility of blood vessels sprouting from the bioartificial graft. Our results showed that cells within the graft were viable and vascular tubes were present at week 4, while cells in a fat tissue block were dead in vitro. In addition, capillaries were observed sprouting from the graft into the Matrigel, demonstrating its angiogenic potential. We expect that improved cell viability and angiogenesis in the bioartificial substitute, compared to intact autologous graft, could potentially contribute to its survival following implantation. Copyright © 2012 John Wiley & Sons, Ltd.

Quantitative MRI Analysis of Salivary Glands in Sickle Cell Disease

Objectives The purpose of this prospective study was to characterize the MR relaxometric features of the major salivary glands in patients with sickle cell disease (SCD). Methods 15 patients with SCD (aged 19.8-43.6 years) and 12 controls were imaged with the mixed turbo-spin echo pulse sequence. The major salivary glands were manually segmented and T(1), T(2) and secular T(2) relaxometry histograms were modelled with Gaussian functions. Results Shortened T(1) relaxation times were seen solely in the submandibular glands of patients with SCD (747.5 ± 54.8 ms vs 807.1 ± 38.3 ms, p < 0.001). Slight T(2) and secular T(2) shortening were seen in the parotid gland; however, this difference was not significant (p = 0.07). The sublingual gland showed no changes under MR relaxometry. There was no difference in glandular volumes, and no correlation was demonstrated between history of blood transfusion and salivary gland relaxometry. Conclusions Patients with SCD exhibited changes in quantitative MRI T(1) relaxometry histograms of the submandibular glands.

A Novel Preparation Method for Silicone Oil Nanoemulsions and Its Application for Coating Hair with Silicone

Silicone oil, as a major component in conditioner, is beneficial in the moisture preservation and lubrication of hair. However, it is difficult for silicone oil to directly absorb on the hair surface because of its hydrophobicity. Stable nanoemulsions containing silicone oil may present as a potential solution to this problem.

High Tolerance to Salinity and Herbivory Stresses May Explain the Expansion of Ipomoea Cairica to Salt Marshes

Invasive plants are often confronted with heterogeneous environments and various stress factors during their secondary phase of invasion into more stressful habitats. A high tolerance to stress factors may allow exotics to successfully invade stressful environments. Ipomoea cairica, a vigorous invader in South China, has recently been expanding into salt marshes.

Psychometric Assessment of the Chinese Version of the Decisional Conflict Scale in Chinese Women Making Decision for Breast Cancer Surgery

OBJECTIVE: The decisional conflict scale (DCS) measures the perception of uncertainty in choosing options, factors contributing to decision conflict and effective decision making. This study examined the validity and reliability of the Chinese version of the DCS in Hong Kong Chinese women deciding breast cancer (BC) surgery. METHOD: A Chinese version of the 16-item DCS was administered to 471 women awaiting initial consultation for BC diagnosis. Confirmatory factor analysis (CFA) assessed the factor structure. Internal consistency, and convergent and discriminant validities of the factor structure were assessed. RESULTS: CFA revealed the original factor structure of the DCS showed poor fit to this sample. Exploratory factor analysis revealed an alternative three-factor structure, Informed and Values Clarity, Uncertainty and Effective Decision and Support, was optimal. Cronbach's alpha ranged from 0.51 to 0.87. Correlations between decision-making difficulties and satisfaction with medical consultation demonstrated acceptable convergent validity. Construct validity was supported by correlations between decision regret and psychological distress. Discriminant validity was supported by differentiation between delaying and non-delaying decision-makers. CONCLUSIONS: The three-factor DCS-14 is a valid and practical measure for assessing decisional conflict in deciding BC surgery. It shows good potential for use in assessing decision satisfaction for women diagnosed with BC.

Use of Radiation Therapy in the Last 30 Days of Life Among a Large Population-Based Cohort of Elderly Patients in the United States

PURPOSE:Our goal was to evaluate use and associated costs of radiation therapy (RT) in the last month of life among those dying of cancer. METHODS:We used the Surveillance, Epidemiology, and End Results (SEER) -Medicare linked databases to analyze claims data for 202,299 patients dying as a result of lung, breast, prostate, colorectal, and pancreas cancers from 2000 to 2007. Logistic regression modeling was used to conduct adjusted analyses of potential impacts of demographic, health services, and treatment-related variables on receipt of RT and treatment with greater than 10 days of RT. Costs were calculated in 2009 dollars. RESULTS: Among the 15,287 patients (7.6%) who received RT in the last month of life, its use was associated with nonclinical factors such as race, gender, income, and hospice care. Of these patients, 2,721 (17.8%) received more than 10 days of treatment. Nonclinical factors that were associated with greater likelihood of receiving more than 10 days of RT in the last 30 days of life included: non-Hispanic white race, no receipt of hospice care, and treatment in a freestanding, versus a hospital-associated facility. Hospice care was associated with 32% decrease in total costs of care in the last month of life among those receiving RT. CONCLUSION:Although utilization of RT overall was low, almost one in five of patients who received RT in their final 30 days of life spent more than 10 of those days receiving treatment. More research is needed into physician decision making regarding use of RT for patients with end-stage cancer.

Tumor Associated Macrophages Regulate Murine Breast Cancer Stem Cells Through a Novel Paracrine EGFR/Stat3/Sox-2 Signaling Pathway

The cancer stem cell (CSC) hypothesis has gained significant recognition as a descriptor of tumorigenesis. Additionally, tumor-associated macrophages (TAMs) are known to promote growth and metastasis of breast cancer. However, it is not known whether TAMs mediate tumorigenesis through regulation of breast CSCs. Here, we report that TAMs promote CSC-like phenotypes in murine breast cancer cells by upregulating their expression of Sox-2. These CSC-like phenotypes were characterized by increased Sox-2, Oct-4, Nanog, AbcG2 and Sca-1 gene expression, in addition to increased drug--efflux capacity, resistance to chemotherapy and increased tumorigenicity in vivo. Downregulation of Sox-2 in tumor cells by siRNA blocked the ability of TAMs to induce these CSC-like phenotypes and inhibited tumor growth in vivo. Furthermore, we identified a novel EGFR/Stat3/Sox-2 paracrine signaling pathway between macrophages and mouse breast cancer cells that is required for macrophage-induced upregulation of Sox-2 and CSC phenotypes in tumor cells. We showed that this cross-talk was effectively blocked by the small molecule inhibitors AG1478 or CDDO-Im against EGFR and Stat3, respectively. Therefore, our report identifies a novel role for TAMs in breast CSC regulation and establishes a rationale for targeting the EGFR/Stat3/Sox-2 signaling pathway for cancer stem cell therapy.

Quantitative Proteomics by Amino Acid Labeling in Foot-and-Mouth Disease Virus (FMDV)-Infected Cells

Foot-and-mouth disease virus (FMDV) is an important disease agent that can be difficult to effectively eradicate from herds. Because it is an obligate intracellular parasite, the virus has multiple effects on the host cell during infection. Here, a high-throughput quantitative proteomic approach was used to develop an unbiased holistic overview of the protein changes in IBRS-2 cells infected with FMDV. Stable isotope labeling with amino acids in cell culture (SILAC) combined with LC-MS/MS was performed to identify and quantify 1260 cellular and 2 viral proteins after 6 h of infection of IBRS-2 cells with FMDV. Of these identified and measured cellular protein pairs, 77 were significantly up-regulated, and 50 were significantly down-regulated based on significance B ≤ 0.05. The differentially altered proteins included a number of proteins involved in endolysosomal proteases system, cell cycle, cellular growth and proliferation, and immune cell trafficking. Selected data were validated by Western blot. Ingenuity Pathway Analysis revealed that proteins that changed in response to infection could be assigned to defined canonical pathways and functional groupings, such as integrin signaling. The obtained data might not only improve the understanding of the dynamics of FMDV and host interaction but may also help elucidate the pathogenic mechanism of FMDV infection.

Novel Cancerization Marker, TP53, and Its Role in Distinguishing Normal Tissue Adjacent to Cancerous Tissue from Normal Tissue Adjacent to Benign Tissue

ABSTRACT: BACKGROUND: The histopathological and molecular heterogeneity of normal tissue adjacent to cancerous tissue (NTAC) and normal tissue adjacent to benign tissue (NTAB), and the availability of limited specimens make deciphering the mechanisms of carcinogenesis challenging. Our goal was to identify histogenetic biomarkers that could be reliably used to define a transforming fingerprint using RNA in situ hybridization. METHODS: We evaluated 15 tumor-related RNA in situ hybridization biomarkers using tumor microarray and samples of seven tumor-adjacent normal tissues from 314 patients. Biomarkers were determined using comprehensive statistical methods (significance of support vector machine-based artificial intelligence and area under curve scoring of classification distribution). RESULTS: TP53 was found to be a most reliable index (P <10-7; area under curve >87%) for distinguishing NTAC from NTAB, according to the results of a significance panel (BCL10, BECN1, BRCA2, FITH, PTCH11 and TP53). CONCLUSIONS: The genetic alterations in TP53 between NTAC and NTAB may provide new insight into the field of cancerization and tumor transformation.

Association of Genetic Variations in GNB1 with Response to Peginterferon Plus Ribavirin Therapy for Chronic Hepatitis C in a Chinese Population in Taiwan

ABSTRACT: BACKGROUND: The aim of this study was to evaluate whether polymorphisms in the guanine nucleotide binding (G protein), beta polypeptide 1 (GNB1) gene are associated with a rapid virological response (RVR) among HCV genotype 1 (HCV-1) and 2 (HCV-2) infected patients receiving peginterferon plus ribavirin treatment (PEG-IFNalpha-RBV) METHODS: We analyzed the association between RVR to PEG-IFNalpha-RBV therapy and 4 tagging single nucleotide polymorphisms (SNPs) of the GNB1 gene. This study included 265 HCV-1 and 195 HCV-2 infected patients in a Chinese population in Taiwan. RESULTS: Among the GNB1 SNPs examined, the combination of genotypes G/G and G/T populations of rs12126768 was significant inversely correlated with RVR in HCV-1 infected patients (P = 0.0330), whereas HCV-2 infected patients, combination of A/A and A/C genotypes populations at rs4648727 responded better to the PEG-IFNalpha-RBV treatment (P = 0.0089). However, there were no significant differences in the allele frequencies of those SNPs between RVR responders and non-responders. Several RVR susceptibility GNB1 haplotypes were identified, and the ACAT haplotype of the 4 SNPs may increase the successful outcomes of HCV-1 and HCV-2 infected patients (P = 0.0261 and P = 0.0253, respectively). CONCLUSION: The data for GNB1 SNPs and the association of RVR showed that GNB1 polymorphisms might be associated with the therapeutic outcomes of HCV-1 and HCV-2 infected patients under standard of care (SOC) treatment.

The γ-Secretase Blocker DAPT Reduces the Permeability of the Blood-Brain Barrier by Decreasing the Ubiquitination and Degradation of Occludin During Permanent Brain Ischemia

BACKGROUND: Tight junction protein degradation is a principal characteristic of the blood-brain barrier (BBB) damage that occurs during brain ischemia. AIMS: We investigated the mechanisms of occludin degradation that underlie permanent middle cerebral artery occlusion (pMCAO) in rats. METHODS AND RESULTS: Western blot and Co-immunoprecipitation data indicated ubiquitination and degradation of occludin in brain after pMCAO, which was consistent with ZO-1 degradation in penumbra regions as observed at 24 h after pMCAO. We further investigated candidate protease(s) responsible for the degradation of occludin during pMCAO. The intraventricular administration of γ-secretase blocker DAPT significantly inhibited the pMCAO-induced neurovascular damage, whereas ALLM and Batimastat, which are inhibitors of calpain and metalloproteinase proteases, respectively, were less effective. Notably, we found that DAPT significantly inhibited BBB disruption in comparison with vehicle treatment, as assessed by Evans blue excretion. Interestingly, the confocal immunostaining revealed that activation of the E3 ubiquitin ligase Itch is associated with degradation of occludin in brain microvessels following ischemia. Furthermore, our data demonstrate that the inhibition of γ-secretase signaling and the itch-mediated ubiquitination of occludin likely underlie the vasoprotective effect of DAPT after pMCAO. CONCLUSION: The γ-secretase blocker DAPT reduces the permeability of the BBB by decreasing the ubiquitination and degradation of occludin during permanent brain ischemia, suggesting that γ-secretase may represent a novel therapeutic target for preventing neurovascular damage.

An Analysis of the Content and Clinical Implications of Online Advertisements for Female Genital Cosmetic Surgery

Women who are contemplating any form of female genital cosmetic surgery (FGCS) are likely to seek information from provider websites. The aim of this study is to examine the breadth, depth and quality of clinical information communicated to women on 10 popular sites and to discuss the implications of the results.

A Novel Amperometric Immunosensor Constructed with Gold-platinum Nanoparticles and Horseradish Peroxidase Nanoparticles As Well As Nickel Hexacyanoferrates Nanoparticles

In this study, three nano-materials comprising gold-platinum nanoparticles (Au-PtNPs), horseradish peroxidase nanoparticles (HRPNPs) and nickel hexacyanoferrate nanoparticles (NiHCFNPs) were used to construct a signal-off immunosensor. Au-PtNPs and NiHCFNPs were assembled on a glass carbon electrode (GCE) by electrodeposition and gold-cyanide bond formation, respectively; anti-fetoproteins (anti-AFP) were immobilized on the Au-PtNPs. HRPNPs were employed to block the possible remaining active sites and avoid nonspecific adsorption. Here, NiHCFNPs served as redox probes, while Au-PtNPs and HRPNPs were used for the synergistic catalysis of H(2)O(2) to amplify the signal. With more and more immunocomplex produced by the antibody-antigen reaction covering the biosensing surface, thus hindering electron transfer, the catalytic peak current will decrease quantitatively in relation to the concentration of target antigen. The resulting immunosensors exhibited a fast response and excellent sensitivity to α-fetoprotein (AFP), and showed two linear ranges in the concentration ranges of 0.06-13 ng mL(-1) and 13-200 ng mL(-1) with a detection limit of 0.017 ng mL(-1).

Benzyl Isothiocyanate Induces Protective Autophagy in Human Prostate Cancer Cells Via Inhibition of MTOR Signaling

Benzyl isothiocyanate (BITC) is a dietary chemopreventive agent that inhibits the growth of various human cancer cells by causing apoptotic cell death. In the present study, we demonstrate that BITC not only induces apoptosis but also induces autophagy in human hormone-sensitive (Rv1) and -refractory (PC3) prostate cancer cells. In BITC-treated cells, the induction of autophagy was detected by monitoring the processing of an autophagy marker protein, LC3, the aggregation of LC3 into granular structures, and the formation of acidic organelles. Inhibition of autophagy using 3-methyladenine (3-MA) increased BITC-induced apoptosis while the administration of caspase inhibitor suppressed BITC-induced cell death. Our data also showed that BITC inhibits mTOR kinase activity in a dose-dependent manner. The expression of phospho-mTOR (Ser2481), an indicator of mTOR intrinsic catalytic activity, and phospho-ULK1 (Ser757), a direct substrate of mTOR, were decreased in BITC-treated cells. However, the increased expression of phospho-mTOR (Ser2448), phospho-AKT (Ser473) and anti-apoptotic Bcl-2 were detected only in PC3 cells at later stages of BITC treatment. Collectively, our results show that BITC induces a protective autophagy response in Rv1 and PC3 cells through inhibition of the mTOR signaling pathway. Activation of the AKT survival pathway was only observed in PC3 cells, representing a resistance mechanism of advanced prostate cancer upon BITC treatment. These findings could potentially contribute to the beneficial effect of BITC in prostate cancer treatments.

[Clinical Study of Treating Knee Osteoarthritis (Bi Syndrome of Knee) by Massage Combined Chinese Materia Medica Footbath Fumigation and Washing]

To evaluate the clinical efficacy of treating knee osteoarthritis (KOA, Bi syndrome of knee) by massage combined Chinese materia medica (CMM) footbath fumigation and washing, and to observe the changes of the Lysholm knee score (LKSS).

[Analysis of GRACE Principle for Comparative Effectiveness Research]

Comparative effectiveness research (CER) now is a very popular concept in the field of international heath care reformation. Although its contents are not essentially changed, it advocates a new medical idea, a policy orientation at the national medical system level. The European countries and America hope CER could initiate the construction of a new historical milestone. Although there are already some guidelines for the design and report of CER by authorized international organizations, its assessment standards have not been involved. Therefore, good research for comparative effectiveness (GRACE) was signed by International Society Pharmacoepidemiology. A series of standard rules were formulated on how to assess the observational studies. In this article by analyzing the GRACE, we hope to provide the referential standards for enforcing observational studies by introducing CER in the clinical studies of Chinese medicine.

[Langerhans Cell Histiocytosis: a Case Report]

A case of Langerhans cell histiocytosis of the mandible was reported. A 34-year-old woman presented with pain for one year on the gingival of the low left jaw, and together with pyorrhea in the past 2 months. The histopathology and immunohistochemical examination confirmed the diagnosis of Langerhans cell histiocytosis.

[Experimental Study on the Best Concentration of SVFs for Promoting Survival Rate of Fat Graft]

To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.

Quantitative Reduction of Peripheral CD4(+) CD25(+) FOXP3(+) Regulatory T Cells in Reproductive Failure After Artificial Insemination by Donor Sperm

PROBLEM: The objective of this study was to determine whether peripheral Treg-cell percentages were altered in women with reproductive failure after artificial insemination by donor sperm (AID) and which parameters can best discriminate women with AID failure and normal controls. METHOD OF STUDY: A retrospective case-control study of 20 fertile controls and 20 patients undergoing more than four treatment cycles with negative pregnancy test (Group I), 20 experiencing biochemical pregnancy loss (Group II), and 20 undergoing missed abortion or spontaneous miscarriage (Group III) was performed. The peripheral percentages of CD4(+) CD25(+) and CD4(+) CD25 (+) Foxp3(+) Treg cells within CD4(+) T-cell population were evaluated at both late follicular and luteal phases of menstrual cycle by flow cytometry. RESULTS: A significantly decreased percentage of CD4(+) CD25(+) Foxp3(+) Treg cells was detected at the late follicular phase in all AID failure groups compared with the controls. The percentage of CD4(+) CD25(+) Foxp3(+) Treg cells at the late follicular phase in the controls was higher than that at the luteal phase. Using receiver operating characteristic curve, we found that CD4(+) CD25(+) Foxp3(+) Treg-cell percentage <2% can best discriminate the AID failure and normal controls. CONCLUSION: Reduced percentage of peripheral CD4(+) CD25(+) Foxp3(+) Treg cells at the late follicular phase was associated with AID failure and can be a potential biomarker for predicting AID-induced failure.

Isolation and Phylogenetic Analysis of Orf Virus from the Sheep Herd Outbreak in Northeast China

ABSTRACT: BACKGROUND: Orf is a zoonotic and epitheliotrophic contagious disease that mainly affects sheep, goats, wild ruminants, and humans with a worldwide distribution. To date, there is little information on the characterization of ORFV strains that are endemic in Mainland China. In addition, the relationship between the severity of disease and the molecular profile of ORFV strains has not been fully elucidated. RESULTS: From the recent outbreak of a sheep herd in Nongan, northeast of China, the novel orf virus (ORFV) strain NA1/11 was successfully isolated. Western blot analysis indicated that the NA1/11 strain cross reacts with monoclonal antibody A3 and infected sheep ORFV antiserum. The purified virions revealed the typical ovoid shape when observed by atomic force microscopy. To determine the genetic characteristics of the NA1/11 strain, the sequences of ORFV011 (B2L), ORFV059 (F1L), ORFV109, ORFV110 and ORFv132 (VEGF) genes were amplified and compared with reference parapoxvirus strains. Non-metric multidimensional scaling (nMDS) was performed to analyze the nucleotide similarities between different ORFV strains. CONCLUSIONS: Phylogenetic analysis based on ORFV 011 nucleotide sequences showed that the NA1/11strain was closely related to Xinjiang and Gansu strains. ORFV110 and ORFV132 genes are highly variable. The results revealed that precise phylogenetic analysis might provide evidence for genetic variation and movement of circulating ORFV strains in Northeast China. In addition, NMDS analysis showed that geographic isolation and animal host are likely major factors resulting in genetic differences between ORFV strains.

Simultaneous Detection, Genotyping and Quantification of Human Papillomaviruses by Multicolor Real-Time PCR and Melting Curve Analysis

Long-term infection of high-risk human papillomavirus (HPV) is the leading cause of cervical cancer while infection of the low-risk HPV is the major reason for condyloma acuminata. An accurate, rapid and convenient assay that is able to simultaneously detect, genotype and quantify HPV is of great clinical value and yet remains to be achieved. We developed a three-color real-time PCR assay that is able to analyze 30 predominant HPVs types in three reactions. The amplification curves indicated the presence of HPV, melting curve analysis identified the HPV genotype, and C(q) value determined the quantity. We applied this assay to 647 cervical swab samples and the results were compared with a commercial genotyping system. The proposed assay had a limit of detection of 5∼50 copies per reaction and a dynamic range of 5×10(1)∼5×10(6) copies per reaction. Comparison study showed that the overall sample concordance with the comparison method was 91.6% and the type agreement was greater than 98.7%. Quantification study demonstrated that the viral loads of HPV 16 in 30 samples with cervical intraepithelial neoplasia (CIN) III lesions were significantly higher than those with CIN I lesions or CIN II lesions and the results were concordant with the comparison method. The increased information content, high throughput, and low cost would facilitate the use of this real-time PCR-based assay in a variety of clinical settings.

Antigenicity and Immunogenicity of RV144 Vaccine AIDSVAX Clade E Envelope Immunogen is Enhanced by a Gp120 N-terminal Deletion

An immune correlates analysis of the RV144 HIV-1 vaccine trial revealed that antibody responses to the gp120 V1V2 region correlated inversely with infection risk. The RV144 protein immunogens (A244-rp120, MN-rgp120) were modified by an N-terminal 11 amino-acid deletion (Δ11) and addition of a HSV (Herpes simplex virus)-gD protein derived tag (gD). We investigated the effects of these modifications on gp120 expression, antigenicity and immunogenicity by comparing unmodified A244 gp120 with both Δ11 deletion and gD tag and with Δ11 only. Analysis of A244 gp120, with or without Δ11 or gD, demonstrated that the Δ11 deletion, without the addition of gD, was sufficient for enhanced antigenicity to gp120 C1 region, conformational V2 and V1/V2 gp120 conformational epitopes. RV144- vaccinee sera IgG bound more avidly to A244 gp120 Δ11 than to the unmodified gp120 and their binding was blocked by C1, V2 and V1/V2 antibodies. Rhesus macaques immunized with the three different forms of A244 gp120 proteins gave similar levels of gp120 antibody titers, although higher antibody titers developed earlier in A244 Δ11 gp120 immunized animals. Conformational V1/V2 mAbs gave significantly higher levels of blocking of plasma IgG from A244 Δ11 gp120 immunized animals than IgG from animals immunized with unmodified A244 gp120, thus indicating a qualitative difference in the V1/V2 antibodies induced by A244 Δ11 gp120. These results demonstrate that deletion of N-terminal residues in the RV144 A244 gp120 immunogen improves both envelope antigenicity and immunogenicity.

Preconfiguration of the Antigen-binding Site During Affinity Maturation of a Broadly Neutralizing Influenza Virus Antibody

Affinity maturation refines a naive B-cell response by selecting mutations in antibody variable domains that enhance antigen binding. We describe a B-cell lineage expressing broadly neutralizing influenza virus antibodies derived from a subject immunized with the 2007 trivalent vaccine. The lineage comprises three mature antibodies, the unmutated common ancestor, and a common intermediate. Their heavy-chain complementarity determining region inserts into the conserved receptor-binding pocket of influenza HA. We show by analysis of structures, binding kinetics and long time-scale molecular dynamics simulations that antibody evolution in this lineage has rigidified the initially flexible heavy-chain complementarity determining region by two nearly independent pathways and that this preconfiguration accounts for most of the affinity gain. The results advance our understanding of strategies for developing more broadly effective influenza vaccines.

Prospective Study of Family History and Colorectal Cancer Risk by Tumor LINE-1 Methylation Level

BackgroundBeyond known familial colorectal cancer (CRC) syndromes, the mechanisms underlying the elevated CRC risk associated with CRC family history remain largely unknown. A recent retrospective study suggests familial clustering of CRC with hypomethylation in long interspersed nucleotide element 1 (LINE-1). We tested the hypothesis that CRC family history might confer a higher risk of LINE-1 methylation-low CRC.MethodsUsing the Nurses' Health Study and the Health Professionals Follow-up Study, we prospectively examined the association between CRC family history and the risk of rectal and colon cancer (N = 1224) according to tumor LINE-1 methylation level by duplication method Cox proportional hazards regression. We examined microsatellite instability (MSI) status to exclude the influence of Lynch syndrome. All statistical tests were two-sided.ResultsThe association between CRC family history and non-MSI CRC risk differed statistically significantly by LINE-1 methylation level (P (heterogeneity) = .02). CRC family history was associated with a statistically significantly higher risk of LINE-1 methylation-low non-MSI cancer (multivariable hazard ratio [HR] = 1.68, 95% confidence interval [CI] = 1.19 to 2.38 for 1 vs 0 first-degree relatives with CRC; multivariable HR = 3.48, 95% CI = 1.59 to 7.6 for ≥2 vs 0 first-degree relatives with CRC; P (trend) < .001). In contrast, CRC family history was not statistically significantly associated with LINE-1 methylation-high non-MSI cancer (P (trend) = .35).ConclusionsThis molecular pathological epidemiology study shows that CRC family history is associated with a higher risk of LINE-1 methylation-low CRC, suggesting previously unrecognized heritable predisposition to epigenetic alterations. Additional studies are needed to evaluate tumor LINE-1 methylation as a molecular biomarker for familial cancer risk assessment.

Facile Spectrophotometric Assay of Molar Equivalents of N-hydroxysuccinimide Esters of Monomethoxyl Poly-(ethylene Glycol) Derivatives

ABSTRACT: BACKGROUND: A new method is developed to quantify molar equivalents of N-hydroxysuccinimide (NHS) esters of derivatives of monomethoxyl poly-(ethylene glycol) (mPEG) in their preparations with NHS acetate ester as the reference. RESULTS: NHS ester of succinic monoester or carbonate of mPEG of 5,000 Da was synthesized and reacted with excessive ethanolamine in dimethylformamide at 25 0C for 15 min. Residual ethanolamine was subsequently quantified by absorbance at 420 nm after reaction with 2,4,6-trinitrobenzenesulfonic acid (TNBS) at pH 9.2 for 15 min at 55 0C followed by cooling with tap water. Reaction products of ethanolamine and NHS esters of mPEG caused no interference with TNBS assay of residual ethanolamine. Reaction between ethanolamine and NHS acetate ester follows 1:1 stoichiometry. By the new method, molar equivalents of NHS esters of carbonate and succinic monoester of mPEG in their preparations were about 90% and 60% of their theoretical values, respectively. During storage at 370C in humid air, the new method detected spontaneous hydrolyses of the two NHS esters of mPEG more sensitively than the classical spectrophotometric method based on absorbance at 260 nm of NHS released by reaction with ammonia in aqueous solution. CONCLUSION: The new method is favorable to quantify molar equivalents of NHS esters of mPEG derivatives and thus control quality of their preparations.

Letter to the Editor: Regarding "Prospective Analysis of Association Between Use of Statins or Other Lipid-Lowering Agents and Colorectal Cancer Risk"

Inactivation of Bacillus Subtilis Spores by Combining High-pressure Thermal Sterilization and Ethanol

High-pressure thermal sterilization (HPTS) is a new and promising sterilization technology of foods. Effects of combining HPTS and ethanol treatment on inactivation of Bacillus subtilis spores were investigated. An interesting phenomenon was observed. The inactivation effect of HPTS treatment on the spores was enhanced significantly with the increase in ethanol concentration from 0 to 15%. However, the inactivation effect was decreased with further increase in ethanol concentration up to 70%. In addition, the release of DPA and leakages of OD(260) and OD(280) material from the spores increased continuously with the increase in ethanol concentration. Moreover, flow cytometry analysis suggested that although the inner membrane of the spores was damaged, PI could not bind with the spore DNA immediately after HPTS treatment. In conclusion, the mechanism of this special phenomenon could be attributed to the germination of spores under HPTS treatment and effects of ethanol on the protein or water activity. HPTS caused other lethal damages to the spores besides its damage to the inner membrane. Ethanol of low concentrations could significantly enhance the sterilization effects of HPTS, which was good for keeping the qualities of foods.

The Associations Between Serum Perfluorinated Chemicals and Thyroid Function in Adolescents and Young Adults

Perfluorinated chemicals (PFCs) have been widely used in a variety of products worldwide for years. However, the effect of PFCs on thyroid function has not yet been clearly defined. We recruited 567 subjects (aged 12-30 years) in a population-based cohort of adolescents and young adults with abnormal urinalysis in the childhood to determine the relationship between serum level of PFCs and the levels of serum free thyroxine (T4) and thyroid stimulating hormone (TSH). The geometric means and geometric standard deviation concentrations of perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), perfluorononanoic acid (PFNA) and perfluoroundecanoic acid (PFUA) were 2.67 (2.96)ng/ml, 7.78 (2.42)ng/ml, 1.01 (3.48)ng/ml and 5.81 (2.92)ng/ml, respectively. Differences in the levels of free T4 and TSH across different categories of PFOA, PFOS and PFUA were insignificant. After controlling for confounding factors, multiple linear regression analyses revealed mean serum level of free T4 increased significantly across categories (<60th, 60-89 and >90th percentiles) of PFNA (P for trend =0.012 in the full model). The association between PFNA and free T4 was more significant in male subjects in age group 20-30, active smokers and in those with higher body mass index in stratified analysis. Serum concentrations of PFNA were associated with serum free T4 levels in adolescents and young adults.

Comparative Performance of C-MAC Video Laryngoscope and Macintosh Direct Laryngoscope for Emergency Intubation

Therapeutic Effect of Autologous Bone Marrow-Derived Liver Stem Cells Transplantation in Hepatitis B Virus-Induced Liver Cirrhosis

Background/Aims: To determine the safety, feasibility and therapeutic effect of in vitro-expanded autologous bone marrow-derived liver stem cells (BMDLSC) transplantation in cirrhotic patients following chronic hepatitis B virus infection. Methodology: Twelve patients with post-hepatitic cirrhosis and portal hypertension who required splenectomy with periesophagogastric devascularization were included and were divided into two groups. Group I included six patients who received autologous BMDLSC infusion via the hepatic artery after in vitro expansion for 7 days. Group II (control group) included six patients who received normal saline infusion via the same route during splenectomy with periesophagogastric devascularization. The therapeutic effects were compared 3 months later. Patients in Group I were followed-up for 24 months after transplantation. Results: There were no adverse effects during short- and long-term follow-ups. Three months after the operation, patients who received BMDLSC infusion showed better hepatic function than those who received saline infusion (p<0.05). Patients in Group I showed stable liver function parameters with no complications during the 24-month follow-up period. Conclusions: Transplantation of in vitro-expanded autologous BMDLSC via the hepatic artery is a safe and effective treatment for decompensated liver cirrhosis.

Effect of Durations of Wheelchair Tilt-in-space and Recline on Skin Perfusion over the Ischial Tuberosity in People with Spinal Cord Injury

OBJECTIVE: To compare the efficacy of various durations of wheelchair tilt-in-space and recline on enhancing skin perfusion over the ischial tuberosity in people with spinal cord injury (SCI). DESIGN: Repeated measures, intervention and outcomes measure design. SETTING: University research laboratory. PARTICIPANTS: Power wheelchair users with SCI (N=9). INTERVENTIONS: Three protocols of various durations (3 min, 1 min and zero) of wheelchair tilt-in-space and recline were randomly assigned to the participants. Each protocol consisted of a baseline 15 min sitting, a duration of zero to 3 min reclined and tilted, a second 15 min sitting, and a 5 min recovery. The position at the baseline and second sitting was no tilt/recline of the participant and at the reclined and tilted and recovery was at 35° tilt-in-space and 120° recline. MAIN OUTCOME MEASURES: Skin perfusion response to tilt and recline was assessed by laser Doppler and was normalized to mean skin perfusion at the baseline sitting. RESULTS: The results showed that mean skin perfusion during recovery at the 3 min duration was significantly higher compared to the 1 min duration (P<.017), and mean skin perfusion was not significantly different between the 1 min and zero durations (N.S.). Skin perfusion during the second sitting was significantly higher at the 3 min duration compared to the 1 min and zero durations (P<.017). CONCLUSIONS: Our findings suggest that performing the 3 min duration of wheelchair tilt-in-space and recline is more effective than the 1 min duration on enhancing skin perfusion of weight bearing soft tissues.

Spatial Distribution and Sources Identification of Elements in PM(2.5) Among the Coastal City Group in the Western Taiwan Strait Region, China

The main purpose of this study was to investigate the spatial variations of 20 elements (Al, Si, Ti, Ca, Fe, Mg, Cr, Mn, Ni, P, S, K, Cu, Cl, V, Se, Br, As, Zn, and Pb) in PM(2.5) (particle matters≤2.5μm in aerodynamic diameter) in the coastal city group in the Western Taiwan Strait (WTS) region, China during spring 2011. The average PM(2.5) mass concentration at 13 sites was 77.0μg/m(3) and the elemental fraction accounted for about 10-20%. Multivariate analyses (principal component analysis and cluster analysis) and a correlation matrix were used to identify the sources of elements in PM(2.5). The results revealed that the elements originated mainly from traffic emissions, coal combustion, pyrometallurgical processes, and crustal sources. Spatially, the concentrations of elements were generally higher in several rapidly growing locations, and the enrichment factors (EFs) for most elements were much higher at the northern sites than those at the southern sites, suggesting that the air quality in the northern part of the study area was strongly affected by anthropogenic activity. Backward wind trajectory analysis during the sampling period indicated that the concentrations of elements in PM(2.5) in the WTS region were greatly impacted by dust particles transported from Northern China in spring.

Increased Risk of Large Postvoid Residual Urine and Lower Long-Term Success Rate in Frail Elderly After Intravesical Onabotulinumtoxin A Injection for Refractory Idiopathic Detrusor Overactivity

PURPOSE: Intravesical injection of onabotulinumtoxin A (BoNT-A) is effective for idiopathic detrusor overactivity (IDO) refractory to antimuscarinics. However, safety is a major concern, especially for the elderly. We investigate the efficacy and safety of intravesical BoNT-A injections for refractory IDO in frail elderly. MATERIALS AND METHODS: From 2004-2009, 166 patients with urodynamic IDO refractory to previous antimuscarinics for more than 3 months received one intravesical 100U BoNT-A injection. "Frail elderly" was defined as those elder than 65 years with the presence of 3 or more of the following criteria: unintentional weight loss, self-reported exhaustion, weakness, slow walking speed, and low physical activity. Treatment results were assessed using patients' perception of bladder condition (PPBC), voiding diary, urodynamic parameters, and Kaplan-Meier estimates of survival plots. RESULTS: Subjects were 61 frail elderly, 63 elderly without frailty, and 42 younger than 65 years. Large post-void residual (PVR) volume (>150 mL) after BoNT-A injection was significantly higher in the frail elderly group (60.7%) than in the other groups (39.7% and 35.7%, p=0.018). Urinary retention developed in 7(11.5%) frail elderly, 4(6.3%) elderly without frailty, and 1(2.4%) younger subjects (p=0.203); the duration of recovery was significantly longer in the frail elderly. The cumulative success rate was significantly lower in the frail elderly than in the other 2 groups (p=0.009). CONCLUSIONS: Although safety and efficacy were similar between elderly without frailty and younger patients, an increased risk of large PVR and lower long-term success rate in frail elderly were noted after intravesical BoNT-A injection for refractory IDO.

Combination of Aβ Clearance and Neurotrophic Factors As a Potential Treatment for Alzheimer's Disease

There is no effective drug to treat Alzheimer's disease (AD), a neurodegenerative disease affecting an estimated 30 million people around the world. Strongly supported by preclinical and clinical studies, amyloid-beta (Aβ) may be a target for developing drugs against AD. Meanwhile, the fact that localized neuronal death/loss and synaptic impairment occur in AD should also be considered. Neuronal regeneration, which does not occur normally in the mammalian central nervous system, can be promoted by neurotrophic factors (NTFs). Evidence from clinical trials has shown that both Aβ clearance and NTFs are potentially effective in treating AD, thus a new approach combining Aβ clearance and administration of NTFs may be an effective therapeutic strategy.

Toxicokinetics of Tilapia Following High Exposure to Waterborne and Dietary Copper and Implications for Coping Mechanisms

One of the major challenges in assessing the potential metal stress to aquatic organisms is explicitly predicting the internal dose in target organs. We aimed to understand the main sources of copper (Cu) accumulation in target organs of tilapia (Oreochromis mossambicus) and to investigate how the fish alter the process of Cu uptake, depuration, and accumulation (toxicokinetics (TK)) under prolonged conditions. We measured the temporal Cu profiles in selected organs after single and combined exposure to waterborne and dietary Cu for 14 days. Quantitative relations between different sources and levels of Cu, duration of treatment, and organ-specific Cu concentrations were established using TK modeling approaches. We show that water was the main source of Cu in the gills (>94 %), liver (>89 %), and alimentary canal (>86 %); the major source of Cu in the muscle (>51 %) was food. Cu uptake and depuration in tilapia organs were mediated under prolonged exposure conditions. In general, the uptake rate, depuration rate, and net bioaccumulation ability in all selected organs decreased with increasing waterborne Cu levels and duration of exposure. Muscle played a key role in accounting for the rapid Cu accumulation in the first period after exposure. Conversely, the liver acted as a terminal Cu storage site when exposure was extended. The TK processes of Cu in tilapia were highly changed under higher exposure conditions. The commonly used bioaccumulation model might lead to overestimations of the internal metal concentration with the basic assumption of constant TK processes.

Inflammatory Cytokine Measurement Quickly Discriminates Gram-negative from Gram-positive Bacteremia in Pediatric Hematology/oncology Patients with Septic Shock

PURPOSE: We performed a prospective study to evaluate the ability of inflammatory cytokines in discriminating gram-negative from gram-positive bacteremia in septic shock. METHODS: During the study period, the serum inflammatory cytokine levels were measured at the onset of septic shock by flow cytometry in pediatric hematology/oncology patients with septic shock. RESULTS: One hundred episodes of septic shock were enrolled. Of 97 episodes of monomicrobial infection, 73.2 % were caused by gram-negative bacteremia and 26.8 % by gram-positive bacteremia. Interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α were closely related to the pediatric index of mortality 2 (PIM2) score and mortality. However, although the PIM2 score and mortality were comparable, the IL-6, IL-10, and TNF-α levels were significantly higher in patients with gram-negative bacteremia (GNB) than those with gram-positive bacteremia (median levels, pg/mL: IL-6: 784.1 vs. 254.4, P = 0.001; IL-10: 192.2 vs. 19.7, P < 0.001; TNF-α: 4.2 vs. 2.0, P < 0.001). Of the three cytokines, IL-10 was the most useful biomarker for GNB prediction in the derivation cohort and a cutoff value of 50 pg/mL showed a sensitivity of 70.8 % and a specificity of 80.0 %, with a positive predictive value of 89.5 %. When this cutoff value was applied to the validation cohort, the sensitivity, specificity, and positive predictive value were 80.9, 75.0, and 90.5 %, respectively. CONCLUSIONS: Flow cytometry-based inflammatory cytokine measurement is a helpful adjuvant approach for early and quick discrimination of gram-negative from gram-positive bacteremia in pediatric hematology/oncology patients with septic shock which might be useful for evaluating the severity of shock and the selection and/or timely withdrawal or switch of antibiotics.

Characterization of Polyethylene Glycol-grafted Polyethylenimine and Superparamagnetic Iron Oxide Nanoparticles (PEG-g-PEI-SPION) As an MRI-visible Vector for SiRNA Delivery in Gastric Cancer in Vitro and in Vivo

BACKGROUND: Gene therapy is a promising therapeutic method but is severely hampered due to its lack of an ideal delivery system. Therefore, in this study, a nonviral and magnetic resonance imaging (MRI) visible vector, polyethylene glycol-grafted polyethylenimine and superparamagnetic iron oxide nanoparticles (PEG-g-PEI-SPION) was used as a nanocarrier for small interfering RNA (siRNA) delivery in gastric cancer. METHODS: Biophysical characterization of PEG-g-PEI-SPION was systematically analyzed, including size, zeta potential, siRNA condensation capacity, cell viability, transfection efficiency, cellular uptake, and MRI-visible function in vivo. Besides, CD44 variant isoform 6 (CD44v6), a protein marker for metastatic behavior in gastric cancer, and was chose as the target gene to further analyze the siRNA delivery function of PEG-g-PEI-SPION. RESULTS: Under comprehensive analysis, the appropriate N/P ratio of PEG-g-PEI-SPION/siRNA was 10,. and siRNA targeting at human CD44v6 (siCD44v6) transferred by PEG-g-PEI-SPION was effective at downregulating the CD44v6 expression of gastric carcinoma cell line SGC-7901 in vitro. Moreover, knockdown of CD44v6 impaired migrating and invasive abilities of SGC-7901 cells. Furthermore, PEG-g-PEI-SPION was a highly efficient contrast agent for MRI scan in vivo. CONCLUSION: PEG-g-PEI-SPION was a promising nonviral vector with molecular image tracing capacity for cancer gene therapy. And CD44v6 was a potential target gene for the prevention and detection of metastatic behavior in gastric cancer.

Targeting Glucosylceramide Synthase Synergizes with C(6)-Ceramide Nanoliposomes to Induce Apoptosis in NK Leukemia

ABSTRACT Natural killer cell leukemia is characterized by clonal expansion of CD3(-) NK cells and comprises both chronic and aggressive forms. Currently, no effective treatment exists, thus providing a need for identification of novel therapeutics. Lipidomic studies revealed dysregulated sphingolipid metabolism as evidenced by decreased levels of overall ceramide species and increased levels of cerebrosides in leukemic NK cells, concomitant with increased glucosylceramide synthase (GCS) expression. GCS, a key enzyme of this pathway, neutralizes pro-apoptotic ceramide by transfer of a UDP-glucose. Thus, we treated both rat and human leukemic NK cells in combination with: 1) exogenous C(6)-ceramide nanoliposomes in order to target mitochondria and increase physiological pro-apoptotic levels of long chain ceramide, and 2) 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol (PPMP), an inhibitor of GCS. Co-administration of C(6)-ceramide nanoliposomes and PPMP elicited an increase in endogenous long-chain ceramide species, which led to cellular apoptosis in a synergistic manner via the mitochondrial intrinsic cell death pathway in leukemic NK cells.

α(0)-Thalassemia Trait with Normal Red Cell Indices: A Report of Two Cases

Thalassemia screening usually involves cut-off values based on a mean corpuscular volume (MCV) of less than 80 fL or a mean corpuscular hemoglobin (MCH) of less than 27 pg. These strategies are able to detect almost all heterozygous carriers of the α-thalassemia (α-thal) (- -(SEA)) deletion in the Chinese population. However, an exception can occur. We describe an α(0)-thal trait with normal red cell indices in two Chinese individuals.

Organocatalyzed Enantioselective Decarboxylative Stereoablation Reaction for the Construction of 3,3'-Disubstituted Oxindoles Using Beta-Ketoacids and 3-Halooxindoles

An unprecedented method for the construction of optically active 3,3'-disubstituted oxindoles via organocatalyzed decarboxylative stereoablation reaction has been developed. We describe the first asymmetric reaction between β-ketoacids and 3-halooxindoles utilizing an organocatalyst. This method allows for the formation of a variety of 3,3'-disubstituted oxindoles bearing a keto-carbonyl group, which are not easily accessible using other methodologies, in moderate to good yields with high enantioselectivities.

Why is the Molybdenum-substituted Tungsten-dependent Formaldehyde Ferredoxin Oxidoreductase Not Active? A Quantum Chemical Study

Formaldehyde ferredoxin oxidoreductase is a tungsten-dependent enzyme that catalyzes the oxidative degradation of formaldehyde to formic acid. The molybdenum ion can be incorporated into the active site to displace the tungsten ion, but is without activity. Density functional calculations have been employed to understand the incapacitation of the enzyme caused by molybdenum substitution. The calculations show that the enzyme with molybdenum (Mo-FOR) has higher redox potential than that with tungsten, which makes the formation of the Mo(VI)=O complex endothermic by 14 kcal/mol. Following our previously suggested mechanism for this enzyme, the formaldehyde substrate oxidation was also investigated for Mo-FOR using the same quantum-mechanics-only model, except for the displacement of tungsten by molybdenum. The calculations demonstrate that formaldehyde oxidation occurs via a sequential two-step mechanism. Similarly to the tungsten-catalyzed reaction, the Mo(VI)=O species performs the nucleophilic attack on the formaldehyde carbon, followed by proton transfer in concert with two-electron reduction of the metal center. The first step is rate-limiting, with a total barrier of 28.2 kcal/mol. The higher barrier is mainly due to the large energy penalty for the formation of the Mo(VI)=O species.

Effectiveness and Safety of a Therapeutic Vaccine Against Angiotensin II Receptor Type 1 in Hypertensive Animals

Primary hypertension is a chronic disease with high morbidity, and the rate of controlled blood pressure is far from satisfactory, worldwide. Vaccination provides a promising approach for treatment of hypertension and improvement in compliance. Here, the ATRQβ-001 vaccine, a peptide (ATR-001) derived from human angiotensin II (Ang II) receptor type 1 conjugated with Qβ bacteriophage virus-like particles, was developed and evaluated in animal models of hypertension. The ATRQβ-001 vaccine significantly decreased the blood pressure of Ang II-induced hypertensive mice up to 35 mm Hg (143±4 versus 178±6 mm Hg; P=0.005) and that of spontaneously hypertensive rats up to 19 mm Hg (173±2 versus 192±3 mm Hg; P=0.003) and prevented remodeling of vulnerable hypertensive target organs. No obvious feedback activation of circulating or local renin-angiotensin system was observed. Additionally, no significant immune-mediated damage was detected in vaccinated hypertensive and nonhypertensive animals. The half-life of the anti-ATR-001 antibody was 14.4 days, surpassing that of existing chemical drugs. In vitro, the anti-ATR-001 antibody specifically bound to Ang II receptor type 1 and inhibited Ca(2+)-dependent signal transduction events, including protein kinase C-α translocation, extracellular signal-regulated kinase 1/2 phosphorylation (72% decrease; P=0.013), and elevation of intracellular Ca(2+) (68% decrease; P=0.017) induced by Ang II, but without inhibiting Ang II binding to the receptor. In conclusion, the ATRQβ-001 vaccine decreased the blood pressure of Ang II-induced hypertensive mice and spontaneously hypertensive rats effectively through diminishing the pressure response and inhibiting signal transduction initiated by Ang II. Thus, the ATRQβ-001 vaccine may provide a novel and promising method for the treatment of primary hypertension.

Phosphorylation of Elongation Factor 2 by Cyclin A-CDK2 Regulates Its Inhibition by EEF2 Kinase

Protein synthesis is highly regulated via both initiation and elongation. One mechanism that inhibits elongation is phosphorylation of Elongation Factor 2 (eEF2) on threonine 56 (T56) by eEF2 kinase (eEF2K). T56 phosphorylation inactivates eEF2 and is the only known normal eEF2 functional modification. In contrast, eEF2K undergoes extensive regulatory phosphorylations that allow diverse pathways to impact elongation. We describe a new mode of eEF2 regulation and show that its phosphorylation by cyclin A-CDK2 on a novel site, serine 595 (S595), directly regulates T56 phosphorylation by eEF2K. S595 phosphorylation varies during the cell cycle and is required for efficient T56 phosphorylation in vivo. Importantly, S595 phosphorylation by cyclin A-CDK2 directly stimulates eEF2 T56 phosphorylation by eEF2K in vitro, and we suggest that S595 phosphorylation facilitates T56 phosphorylation by recruiting eEF2K to eEF2. S595 phosphorylation is thus the first known eEF2 modification that regulates its inhibition by eEF2K and provides a novel mechanism linking the cell cycle machinery to translational control. Because all known eEF2 regulation is exerted via eEF2K, S595 phosphorylation may globally couple the cell cycle machinery to regulatory pathways that impact eEF2K activity.

Transcriptome and Gene Expression Analysis of the Rice Leaf Folder, Cnaphalocrosis Medinalis

The rice leaf folder (RLF), Cnaphalocrocis medinalis (Guenee) (Lepidoptera: Pyralidae), is one of the most destructive pests affecting rice in Asia. Although several studies have been performed on the ecological and physiological aspects of this species, the molecular mechanisms underlying its developmental regulation, behavior, and insecticide resistance remain largely unknown. Presently, there is a lack of genomic information for RLF; therefore, studies aimed at profiling the RLF transcriptome expression would provide a better understanding of its biological function at the molecular level.

GSK3beta-Mediated Drp1 Phosphorylation Induced Elongated Mitochondrial Morphology Against Oxidative Stress

Multiple phosphorylation sites of Drp1 have been characterized for their functional importance. However, the functional consequence of GSK3beta-mediated phosphorylation of Drp1 remains unclear. In this report, we pinpointed 11 Serine/Threonine sites spanning from residue 634∼736 of the GED domain and robustly confirmed Drp1 Ser693 as a novel GSK3beta phosphorylation site. Our results suggest that GSK3beta-mediated phosphorylation at Ser693 does cause a dramatic decrease of GTPase activity; in contrast, GSK3beta-mediated phosphorylation at Ser693 appears not to affect Drp1 inter-/intra-molecular interactions. After identifying Ser693 as a GSK3beta phosphorylation site, we also determined that K679 is crucial for GSK3beta-binding, which strongly suggests that Drp1 is a novel substrate for GSK3beta. Thereafter, we found that overexpressed S693D, but not S693A mutant, caused an elongated mitochondrial morphology which is similar to that of K38A, S637D and K679A mutants. Interestedly, using H89 and LiCl to inhibit PKA and GSK3beta signaling, respectively, it appears that a portion of the elongated mitochondria switched to a fragmented phenotype. In investigating the biofunctionality of phosphorylation sites within the GED domain, cells overexpressing Drp1 S693D and S637D, but not S693A, showed an acquired resistance to H(2)O(2)-induced mitochondrial fragmentation and ensuing apoptosis, which affected cytochrome c, capase-3, -7, and PARP, but not LC3B, Atg-5, Beclin-1 and Bcl2 expressions. These results also showed that the S693D group is more effective in protecting both non-neuronal and neuronal cells from apoptotic death than the S637D group. Altogether, our data suggest that GSK3beta-mediated phosphorylation at Ser693 of Drp1 may be associated with mitochondrial elongation via down-regulating apoptosis, but not autophagy upon H(2)O(2) insult.

The Mitochondrial DNA Northeast Asia CZD Haplogroup Is Associated with Good Disease-Free Survival Among Male Oral Squamous Cell Carcinoma Patients

Reprogramming of energy metabolism in cancer cells has been directly/indirectly linked to mitochondria and mitochondrial functional defects and these changes seem to contribute to the development and progression of cancer. Studies have indicated that mitochondrial DNA haplogroups are associated with risk in relation to various diseases including cancer. However, few studies have examined the effect of haplogroups on cancer prognosis outcome. In order to explore the role of haplogroups on oral squamous cell carcinoma (OSCC) prognosis, the mitochondrial genomes of 300 male OSCC patients were comprehensively analyzed by direct sequencing. They were then haplotyped and grouped into four major geographic haplogroups, namely the East Asia AN, Southeast Asia RBF, East Asia MGE and Northeast Asia CZD groups. The Kaplan-Meier plot analysis indicated that individuals who were members of the CZD haplogroup showed a significant association with better disease-free survival (DFS) than the other three haplogroups and this phenomenon still existed after adjusting for tumor stage, differentiation and age at diagnosis (hazard ratio = 0.55; 95% CI = 0.36-0.84). In addition, an interaction between membership of the RBF haplogroup and radiotherapy/chemo-radiotherapy in DFS was also identified. The results strongly support the hypothesis that an individual's haplogroup, by defining their genomic background, plays an important role in tumor behavior and mitochondrially-targeted anticancer drugs are promising future therapeutic approaches.

SKF83959, an Agonist of Phosphatidylinositol-Linked D(1)-Like Receptors, Promotes ERK1/2 Activation and Cell Migration in Cultured Rat Astrocytes

Extracellular signal-regulated kinase 1/2 (ERK1/2) is a member of the mitogen-activated protein kinase family. It can mediate cell migration. Classical dopamine receptor-mediated ERK1/2 phosphorylation is widely studied in neurons. Here, we report that ERK1/2 phosphorylation is also modulated by putative phosphatidylinositol-linked D(1)-like receptors in cultured rat astrocytes. 6-chloro-7,8-dihydroxy-3-methyl-1-(3-methylphenyl)-2,3,4,5-tetrahydro-1H-3-benzazepine (SKF83959), an agonist of the putative phosphatidylinositol-linked D(1)-like receptors, was found to enhance ERK1/2 phosphorylation, which then promoted the migration of cultured astrocytes. The SKF83959-induced ERK1/2 phosphorylation was found to be Ca(2+)-independent based on the following observations: i. chelating intracellular Ca(2+) did not inhibit ERK1/2 phosphorylation and astrocyte migration; ii. blockage of the release of intracellular Ca(2+) from the endoplasmic reticulum by an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptor did not attenuate ERK1/2 phosphorylation. However, inhibition of phospholipase C (PLC), the upstream molecule of internal Ca(2+) release, disabled SKF83959's ability to elevate the level of ERK1/2 phosphorylation. Both non-selective protein kinase C (PKC) inhibitor and PKCδ selective inhibitor prevented ERK1/2 phosphorylation increase and astrocyte migration, but PKCα inhibitor did not. This suggests that Ca(2+)-independent and diacylglycerol-dependent PKCδ acts downstream of putative phosphatidylinositol-linked D(1)-like receptor activation and mediates SKF83959-induced elevation of ERK1/2 phosphorylation in order to modulate astrocyte migration. In conclusion, our results demonstrate that SKF83959-induced increases in ERK1/2 phosphorylation and astrocyte migration are dependent on PLC-PKCδ signals. This might help us to further understand the functions of the putative phosphatidylinositol-linked D(1)-like receptors in the nervous system.

The CCL2/CCR2 Axis Enhances Vascular Cell Adhesion Molecule-1 Expression in Human Synovial Fibroblasts

Chemokine ligand 2 (CCL2), also known as monocyte chemoattractant protein-1 (MCP-1), belongs to the CC chemokine family that is associated with the disease status and outcomes of osteoarthritis (OA). Here, we investigated the intracellular signaling pathways involved in CCL2-induced vascular cell adhesion molecule-1 (VCAM-1) expression in human OA synovial fibroblasts (OASFs).

Abundant Microsatellite Diversity and Oil Content in Wild Arachis Species

The peanut (Arachis hypogaea) is an important oil crop. Breeding for high oil content is becoming increasingly important. Wild Arachis species have been reported to harbor genes for many valuable traits that may enable the improvement of cultivated Arachis hypogaea, such as resistance to pests and disease. However, only limited information is available on variation in oil content. In the present study, a collection of 72 wild Arachis accessions representing 19 species and 3 cultivated peanut accessions were genotyped using 136 genome-wide SSR markers and phenotyped for oil content over three growing seasons. The wild Arachis accessions showed abundant diversity across the 19 species. A. duranensis exhibited the highest diversity, with a Shannon-Weaver diversity index of 0.35. A total of 129 unique alleles were detected in the species studied. A. rigonii exhibited the largest number of unique alleles (75), indicating that this species is highly differentiated. AMOVA and genetic distance analyses confirmed the genetic differentiation between the wild Arachis species. The majority of SSR alleles were detected exclusively in the wild species and not in A. hypogaea, indicating that directional selection or the hitchhiking effect has played an important role in the domestication of the cultivated peanut. The 75 accessions were grouped into three clusters based on population structure and phylogenic analysis, consistent with their taxonomic sections, species and genome types. A. villosa and A. batizocoi were grouped with A. hypogaea, suggesting the close relationship between these two diploid wild species and the cultivated peanut. Considerable phenotypic variation in oil content was observed among different sections and species. Nine alleles were identified as associated with oil content based on association analysis, of these, three alleles were associated with higher oil content but were absent in the cultivated peanut. The results demonstrated that there is great potential to increase the oil content in A. hypogaea by using the wild Arachis germplasm.

Silver Conductive Features on Flexible Substrates from a Thermally Accelerated Chain Reaction at Low Sintering Temperatures

In this study, a simple and effective silver ink formulation was developed to generate silver tracks with high electrical conductivity on flexible substrates at low sintering temperatures. Diethanolamine (DEA), a self-oxidizing compound at moderate temperatures, was mixed with silver ammonia solution to form a clear and stable solution. After inkjet-printed or pen-written on plastic sheets, DEA in the silver ink decomposes at temperatures higher than 50oC and generates formaldehyde, which reacts spontaneously with silver ammonia ions to form silver thin films. The electrical conductivity of the inkjet-printed silver films can be 26% of bulk silver after heating at 75 oC for 20 minutes, and show great adhesion on plastic sheets.

Microbial Synthesis of N-butanol, Isobutanol, and Other Higher Alcohols from Diverse Resources

Microbial production of fuel and chemical feedstock is a promising approach to solving energy and environmental problems. n-Butanol, isobutanol and other higher alcohols are of particular interest because they can serve as both fuel and chemical feedstock. Alternative resources such as CO(2), syngas, waste protein, and lignocellulose are currently being investigated for their potential to produce these compounds. Except for lignocellulose, utilization of such alternative resource has not been examined extensively. This review aims to summarize the development of metabolic pathways for efficient synthesis of these higher alcohols and the current status of microbial strain development for the conversion of diverse resources into higher alcohols.

Effect of Initial PH Control on Biological Phosphorus Removal Induced by the Aerobic/extended-idle Regime

Recently, it has been reported that biological phosphorus removal (BPR) can be induced by an aerobic/extended-idle (AEI) regime. This study further investigated the effect of initial pH ranging from 6.6 to 8.2 on BPR in the AEI process, and compared the BPR performance between the AEI and the anaerobic/oxic (A/O) regimes under their optimal initial pH value. Experimental results firstly showed that phosphorus removal linearly increased with initial pH increasing from 6.6 to 7.8, but slightly decreased when initial pH increased from 7.8 to 8.2. The optimal initial pH should be controlled at 7.8, and the phosphorus removal at initial pH 7.8 was approximately 1.7-time than that at initial pH 6.6. The mechanism studies showed that the biomass cultured at initial pH 7.8 contained more polyphosphate accumulating organisms (PAOs), lower glycogen accumulating organisms (GAOs), and had higher activities of exopolyphosphatase and polyphosphate kinase than that cultured at initial pH 6.6. Cyclic studies revealed that initial pH control affected the transformations of intracellular polyhydroxyalkanoates and glycogen, which might thereby influence microbial competition between PAOs and GAOs. Then, BPR performance between the AEI and the A/O regimes by adjusting initial pH at 7.8 was also compared. The results showed the AEI regime could drive a better BPR than the generally accepted A/O regime (98% vs 88%). Finally, controlling initial pH at 7.8 to promote BPR in the AEI process was confirmed for a municipal wastewater.

Spinal MCP-1 Contributes to the Development of Morphine Antinociceptive Tolerance in Rats

: The chemokine monocyte chemoattractant protein-1 (MCP-1) has been shown to contribute to neuropathic pain. However, whether MCP-1 is involved in the development of morphine antinociceptive tolerance is incompletely understood.

Focused Ultrasound Therapy for Vulvar Intraepithelial Neoplasia in a Mice Model

BACKGROUND:: The purpose of this study was to determine the effectiveness and safety of focused ultrasound (FU) for the treatment of vulvar intraepithelial neoplasia (VIN) in a mice model. METHODS:: Estradiol benzoate was subcutaneously injected into the abdomens of eighty 129/J mice. VIN was successfully induced in 56 mice and was divided into the FU group and the control group. Pathologic features and changes in vascular endothelial growth factor expression in the lesions were analyzed before and after treatment. RESULTS:: Two months after treatment, lesions in 25 of the 56 mice showed restoration of normal skin. Nineteen of the 21 VINI and VINII lesions returned to normal and the other 2 VINII lesions were down graded to VINI, yielding a curative rate of 90.1%. In the control group, all 21 mice had persistent VIN (P < 0.0001). In the 14 mice with VINIII lesions, 6 returned to normal skin histology representing a curative rate of 42.9%, 5 were reclassified as VINI and 3 were reclassified as VINII. Thus, the total effectiveness rate was 100%. CONCLUSIONS:: The present study suggests that FU therapy is effective, noninvasive and safe in treating VIN in a mice model.

Modulation of Nanocavity Plasmonic Emission by Local Molecular States of C60 on Au(111)

We investigate the modulation of C60 monolayers on the nanocavity plasmonic (NCP) emission on Au(111) by tunneling electron excitation from a scanning tunneling microscope (STM) tip. STM induced luminescence spectra show not only suppressed emission, but also significant redshift of NCP emission bands on the C60 molecules relative to the bare metal surface. The redshift, together with the bias- and coverage-dependent emission feature, indicates that the C60 molecules act beyond a pure dielectric spacer, their electronic states are heavily involved in the inelastic tunneling process for plasmonic emission. A modified quantum cutoff relation is proposed to explain qualitatively the observed emission feature at both bias polarities. We also demonstrate molecularly resolved optical contrast on the C60 monolayer and discuss the contrast mechanism briefly.

Observation of Unusual Optical Transitions in Thin-film Cu(In,Ga)Se2 Solar Cells

In this paper, we examine photoluminescence spectra of Cu(In,Ga)Se2 (CIGS) via temperature-dependent and power-dependent photoluminescence (PL). Donor-acceptor pair (DAP) transition, near-band-edge transition were identified by their activation energies. S-shaped displacement of peak position was observed and was attributed to carrier confinement caused by potential fluctuation. This coincides well with the obtained activation energy at low temperature. We also present a model for transition from VSe to VIn and to VCu which illustrates competing mechanisms between DAPs recombinations.

[Identification Characters of Leaf Morphological and Venation Pattern of Abutilon Indicum with Its Confused Herb A. Theophrasti]

To study the identification characters of Abutilon indicum and its confused herb A. theophrasti and establish an identification method.

[The Characteristic of T Cells Response to HBV-specific Antigen Proteins in Patients with HBV Infection]

To analyze the characteristic of T cell response to specific antigen proteins in patients with hepatitis B virus infection.

[Evaluation of Computational Methods for HLA Three Loci Haplotype by Compare with Family-based Data]

We evaluated the accuracy and efficiency of computational inference methods for haplotype on estimate HLA-A-B-C haplotype frequencies by compared with the haplotypes manually defined in a family-base dataset.

High-Efficiency Broadband Anomalous Reflection by Gradient Meta-Surfaces

We combine theory and experiment to demonstrate that a carefully designed gradient meta-surface supports high-efficiency anomalous reflections for near-infrared light following the generalized Snell's law, and the reflected wave becomes a bounded surface wave as the incident angle exceeds a critical value. Compared to previously fabricated gradient meta-surfaces in infrared regime, our samples work in a shorter wavelength regime with a broad bandwidth (750-900 nm), exhibit a much higher conversion efficiency (∼80%) to the anomalous reflection mode at normal incidence, and keep light polarization unchanged after the anomalous reflection. Finite-difference-time-domain (FDTD) simulations are in excellent agreement with experiments. Our findings may lead to many interesting applications, such as antireflection coating, polarization and spectral beam splitters, high-efficiency light absorbers, and surface plasmon couplers.

Novel PRRT2 Mutations in Paroxysmal Dyskinesia Patients with Variant Inheritance and Phenotypes

Paroxysmal dyskinesias (PDs) are a group of episodic movement disorders with marked variability in clinical manifestation and potential association with epilepsy. PRRT2 has been identified as a causative gene for PDs, but the phenotypes and inheritance patterns of PRRT2 mutations need further clarification. In this study, ten familial and 21 sporadic cases with PDs and PDs-related phenotypes were collected. Genomic DNA was screened for PRRT2 mutations by direct sequencing. Seven PRRT2 mutations were identified in nine (90.0%) familial cases, and in six (28.6%) sporadic cases.Five mutations are novel: two missense mutations (c.647C>G/p.Pro216Arg and c.872C>T/p.Ala291Val) and three truncating mutations (c.117delA/p.Val41TyrfsX49, c.510dupT/p.Leu171SerfsX3 and c.579dupA/p.Glu194ArgfsX6). Autosomal dominant inheritance with incomplete penetrance was observed in most of the familial cases. In the sporadic cases, inheritance was heterogeneous including recessive inheritance with compound heterozygous mutations, inherited mutations with incomplete parental penetrance and de novo mutation. Variant phenotypes associated with PRRT2 mutations, found in 36.0% of the affected cases, included febrile convulsions, epilepsy, infantile non-convulsive seizures (INCS) and nocturnal convulsions (NC). All patients with INCS or NC, not reported previously, displayed abnormalities on EEG. No EEG abnormalities were recorded in patients with classical ICCA/PKD. Our study further confirms that PRRT2 mutations are the most common cause of familial PDs, displaying both dominant and recessive inheritance. Epilepsy may occasionally occur in ICCA/PKD patients with PRRT2 mutations. Variant phenotypes INCS or NC differ from classical ICCA/PKD clinically and electroencephalographically. They have some similarities with, but not identical to epilepsy, possibly represent an overlap between ICCA/PKD and epilepsy.

Comparative Outcomes of Two Nasoalveolar Molding Techniques for Unilateral Cleft Nose Deformity

: Nasoalveolar molding is increasingly being used to treat unilateral cleft nose deformity before primary repair. The Grayson technique starts nasal molding when an alveolar gap is reduced to 5 mm, whereas the Figueroa technique performs nasal and alveolar molding at the same time. The authors investigated the comparative efficacy, efficiency, and incidence of complications of the two techniques.

Association of Cardiovascular and Metabolic Disease Genes with Psoriasis

Design, Synthesis, Antinociceptive and Anti-inflammatory Activities of Novel Piroxicam Analogues

In this paper we report the design, synthesis, antinociceptive and anti-inflammatory activities of a series of benzothiazine N-acylhydrazones 14a–h, planned by structural modification of piroxicam (1), a non steroidal anti-inflammatory drug. Among the synthesized analogues, compounds 14f (LASSBio-1637) and 14g (LASSBio-1639) were identified as novel antinociceptive and anti-inflammatory prototypes, active by oral administration, acting by a mechanism of action that seems to be different from that of piroxicam, since they were inactive as an inhibitor of cyclooxygenase (COX-1 and COX-2) at concentrations of 10 mM.

Proteomics Dissection of Plant Responses to Mineral Nutrient Deficiency

Plants require at least 17 essential nutrients to complete their life cycle. Except for carbon (C), hydrogen (H) and oxygen (O), other essential nutrients are mineral nutrients, which are mainly acquired from soils by roots. In natural soils, the availability of most essential mineral nutrients is very low and hard to meet the demand of plants. Developing crops with high nutrient efficiency is essential for sustainable agriculture, which requires more understandings of crop responses to mineral nutrient deficiency. Proteomic techniques provide a crucial and complementary tool to dissect molecular mechanisms underlying crop adaptation to mineral nutrient deficiency in the rapidly processing post-genome era. This review gives a comparative overview about identification of mineral nutrient deficiency responsive proteins using proteomic analysis, and discusses the current status for crop proteomics and its challenges to be integrated into systems biology approaches for developing crops with high mineral nutrient efficiency.

Clinical Efficacy and Safety of Buyang Huanwu Decoction for Acute Ischemic Stroke: a Systematic Review and Meta-analysis of 19 Randomized Controlled Trials

Buyang Huanwu Decoction (BHD) is a well-known traditional Chinese herbal prescription for treating stroke-induced disability. The objective of this study was to evaluate the efficacy and safety of BHD for acute ischemic stroke. A systematic literature search was performed in 6 databases until February 2012. Randomized controlled clinical trials (RCTs) that evaluate efficacy and safety of BHD for acute ischemic stroke were included. Nineteen RCTs with 1580 individuals were identified. The studies were generally of low methodological quality. Only one of the trial included death or dependency as a primary outcome measure. Only 4 trials reported adverse events. Meta-analysis showed the clinical effective rate of neurological deficit improvement favoring BHD when compared with western conventional medicines (WCM), P < 0.001. There is significant difference in the neurologic deficit score between the BHD treatment group and the WCM control group, P < 0.001. In Conclusion, BHD appears to improve neurological deficit and seems generally safe in patients with acute ischemic stroke. However, the current evidence is insufficient to support a routine use of BHD for acute ischemic stroke due to the poor methodological quality and lack of adequate safety data of the included studies. Further rigorously designed trials are required.

Fentanyl Inhibits Progression of Human Gastric Cancer MGC-803 Cells by NF-kappaB Downregulation and PTEN Upregulation in Vitro

Fentanyl is used as an analgesic to treat pain in a variety of patients with cancer. Moreover, fentanyl may affect tumor growth in many cell lines. To gain better insight into the interaction between fentanyl and tumor, we investigated the effects of fentanyl on the growth of gastric carcinoma cells and the expression of some apoptosis-related genes including NF-kappaB and PTEN. A human gastric cancer cell line MGC-803 was used. The viability and proliferation of gastric cancer MGC-803 cells were detected by MTT assay and colony formation assay. The cell cycle progression and apoptosis were assessed by flow cytometry and the ultrastructure of cells was examined with transmission electron microscope. The migration of cells was investigated by wound healing assay. The expression of NF-kappaB and PTEN was evaluated by semiquantitative RT-PCR and Western blot. Our data showed that fentanyl could inhibit cell growth and proliferation and made cell cycle arrest at G2/M phase. Compared with control cells, MGC-803 cells that were incubated with fentanyl also had a higher apoptotic rate. Fentanyl could lead to morphological changes of gastric cancer cells and reduce the motility of MGC-803 cells. Moreover, fentanyl could downregulate NF-kappaB and upregulate PTEN, which might be the mechanism of fentanyl inhibiting gastric cancer progression in vitro.

Taqman Real-time PCR Assay Based on ORFV024 Gene for Rapid Detection of Orf Infection

In this study, a TaqMan real-time PCR assay was developed to detect and quantify orf virus (ORFV) DNA in infected cell culture and clinical samples. Primers and probe were designed to amplify an 87 bp fragment DNA based on the sequence of ORFV024 gene encoding a NF-κB inhibitor of Orf virus. The assay was highly specific and sensitive for ORFV DNA and no cross-reactions were detected with any other poxviruses. The sensitivity was 5fg or 15 copies of ORFV genomic DNA. Both intra- (1.490±1.261%) and inter-assay (1.958±0.568%) variabilities were within the acceptable range indicating the high efficiency and reproducibility of the assay. Further, the assay has shown a relative diagnostic sensitivity and specificity of 100%, when compared to B2L gene based semi-nested PCR. The assay is simple, rapid, specific and sensitive with a wide potential for rapid field diagnosis of orf in sheep and goats.

Establishment of a Bioluminescence-based Bioassay for the Detection of Dioxin-like Compounds

Abstract Dioxin and dioxin-like compounds are among the most prevalent and toxic environmental pollutants. At present, analytical chemical techniques are considered the gold standard for detection of dioxins. Here, we describe a highly sensitive and cost-effective alternative, based on bioluminescence and bioluminescence resonance energy transfer (BRET). Upon binding to dioxin, Aryl hydrocarbon receptor (AHR) dissociates from HSP90 and subsequently translocates to the nucleus, where it interacts with AHR nuclear translocator (ARNT). We generated cell lines that stably co-express a fusion protein of AHR and Renilla luciferase (AHR-RL) and either HSP90 or ARNT tagged with yellow fluorescent protein (HSP90-YFP or ARNT-YFP). The fluorescent signals of YFP are activated by the emission of RL while the interactions between AHR and HSP90 (or ARNT) were monitored. Application of 3-methylcholanthrene (3MC), the AHR agonist, enhances BRET signals in cells co-expressing AHR-RL, AIP-HIS, P23-HIS and ARNT-YFP (AAPA cells), while suppressing BRET signals in cells co-expressing AHR-RL, AIP-HIS, P23-HIS and HSP90-YFP (AAPH cells). In addition, dioxin treatment reduced Renilla luminescence in AAPH cells in a concentration-dependent manner, due to degradation of AHR. Intriguingly, the detection limit for dioxin in our AHR degradation assay was as low as 10(-17) M. This work highlights the potential of AHR-RL degradation assays to detect dioxin-like pollutants.

Thymic Stromal Lymphopoietin Gene Promoter Polymorphisms and Expression Levels in Graves' Disease and Graves' Ophthalmopathy

ABSTRACT: BACKGROUND: Graves disease (GD) is an organ-specific autoimmune disease characterized by hyperthyroidism, diffuse goiter, autoantibodies against thyroid-specific antigens, and dermopathy. Studies of GD have demonstrated the importance of the Th2 and Th17 immune responses in mediating disease progression. In the present study, we investigated the role of a Th2 cytokine, thymic stromal lymphopoietin (TSLP), in GD and Th17 differentiation. METHODS: In this study, we genotyped 470 patients with GD at 3 single nucleotide polymorphisms (SNPs) in TSLP. In addition, the serum concentrations of TSLP were determined in 432 patients and 272 controls. Ten patients and controls each were further screened using in vitro Th17 differentiation assays. The SNPs were genotyped using ABI TaqMan(R) SNP genotyping assays. For the Th17 differentiation assays, peripheral blood mononuclear cells (PBMCs) isolated from the patients and controls were placed into Th17 differentiation media, and interleukin 17 expression levels were determined. RESULTS: Haplotype analysis indicated that patients with the Ht3 (TCC) haplotype have a 3.28-fold higher risk of developing GD (p = 0.007), whereas those with the Ht5 (TCG) haplotype had a 0.03-fold, reduced risk of developing GD (p = 1 x 10-14). SNP rs3806933 (p = 0.007) was associated with female Graves ophthalmopathy (GO). TSLP expression levels were higher in GD patients than in control subjects, and TLSP was also shown to promote the differentiation of Th17 cells in GD patients. CONCLUSIONS: These results suggest that polymorphisms in TSLP may be used as genetic markers for the diagnosis and prognosis of GD. Furthermore, TLSP may be a target for treating GD.

Programmable Cellular Retention of Nanoparticles by Replacing the Synergistic Anion of Transferrin

The ability to program the intracellular retention of nanoparticles (NPs) would increase their applicability for imaging and therapeutic applications. To date, there has been no efficient method developed to control the fate of NPs once they enter cells. Existing approaches to manipulate the intracellular retention of NPs are mostly "passive" and particle size-dependent. Different sized particles hold distinct cellular responses. The adverse effect of particle size may limit the utility of nanodelivery systems. Therefore, the development of tunable/"active" NP intracellular retention systems with fixed particle sizes remains a considerable challenge. By replacing the synergistic anions of transferrin (Tf) immobilized on quantum dots (Tf-QDs, ca. 25 nm), we have examined the feasibility of this concept. Substitution of synergistic anions of Tf from carbonate (holo-Tf) to oxalate (oxa-Tf) significantly increased the intracellular accumulation of the oxa-Tf-QDs as a result of (i) a delay in cellular removal triggered by oxalate (oxa-Tf)-induced endosomal Tf iron-release retardation, and (ii) enhanced recycling of Tf-QD/TfR (Tf receptor) complexes from early endosomes to the plasma membrane. This accumulation extended the intracellular NP retention interval. The half-maximum fluorescence intensity of the oxa-Tf-QDs in vivo was four times higher than that of the holo-Tf-QDs. Programming of the intracellular NP retention time was accomplished through manipulation of the ratio of holo- and oxa-Tfs on the surfaces of the QDs. Using this simple and efficient approach, it was possible to readily achieve a desirable intracellular retention interval for the NPs.

Significance of CD4 T-cell Adenosine Triphosphate Levels Monitoring in Elderly Renal Transplant Recipients

To find the significance of CD4 T-cell adenosine triphosphate (ATP) levels in elderly renal recipients in correlation with drug doses, levels, and clinical parameters.

Accuracy of In-vitro Tooth Volumetric Measurements from Cone-beam Computed Tomography

The aims of this study were to determine the accuracy of volumetric measurements of teeth in vitro by cone-beam computed tomography (CBCT) and to analyze the factors affecting their volume measurements from the CBCT data.

Fungal Pretreatment of Switchgrass for Improved Saccharification and Simultaneous Enzyme Production

This study investigates fungal pretreatment of switchgrass involving solid state fermentation (SSF) to improve saccharification and simultaneously produce enzymes as co-products. The results revealed that the fungus Pycnoporus sp. SYBC-L3 can significantly degrade lignin and enhance enzymatic hydrolysis efficiency. After a 36-d cultivation period, nearly 30% reduction in lignin content was obtained without significant loss of cellulose and hemicellulose, while a considerable amount of laccase, as high as 6.3U/g, was produced. After pretreatment, pores on switchgrass surface were observed using scanning electron microscopy (SEM). The enzymatic hydrolysis efficiency for the switchgrass with 36-d pretreatment was about 50% greater than the untreated one. Our results suggest that solid state fungal cultivation may be a good method for switchgrass pretreatment, which can simultaneously achieve high efficiency of enzymatic hydrolysis and production of some useful enzymes for other industrial utilization.

Effects of Dairy Manure and Corn Stover Co-digestion on Anaerobic Microbes and Corresponding Digestion Performance

This study investigated the effects of corn stover as a supplemental feed on anaerobic digestion of dairy manure under different hydraulic retention times (HRT). The results elucidated that both HRT and corn stover supplement significantly influenced microbial community and corresponding anaerobic digestion performance. The highest biogas production of 497mL per gram total solid loading per day was observed at a HRT of 40days from digestion of manure supplemented with corn stover. Biogas production was closely correlated with the populations of Bacteroidetes, Clostridia and methanogens. Composition of the solid digestate (AD fiber) from the co-digestion of corn stover and dairy manure was similar to the digestion of dairy manure. However, the hydrolysis of AD fiber was significantly (P<0.05) different among the different digestions. Both HRT and feed composition influenced the hydrolyzability of AD fiber via shifting the composition of microbial community.

Characteristics of Wastewater and Mixed Liquor and Their Role in Membrane Fouling

Effects of wastewater and mixed liquor characteristics on membrane fouling in both a submerged anaerobic membrane bioreactor and a thermophilic submerged aerobic membrane bioreactor were studied with four types of industrial wastewaters. Significant differences in particle size distribution, colloidal content, the protein to polysaccharide ratio, and soluble compounds molecular weight distribution were observed among the four types of wastewaters and mixed liquors. Differences in wastewater and mixed liquor characteristics were correlated to the changes in membrane filtration behavior in both systems. The colloidal content in feed and mixed liquor plays a dominant role and is more important than the quantity of total suspended solids in controlling membrane fouling. The ratio of proteins to polysaccharides is more important than the total quantity of soluble organic substances in controlling membrane fouling. A full characterization of feed and mixed liquor may be used as a tool to predict membrane performance.

Figure-of-merit Enhancement in Nanostructured FeSb(2-x)Ag(x) with Ag(1-y)Sb(y) Nanoinclusions

We present the figure-of-merit (ZT) improvement in nanostructured FeSb(2-x)Ag(x) with Ag(1-y)Sb(y) nanoinclusions through a metal/semiconductor interface engineering approach. Owing to the interfaces between FeSb(2-x)Ag(x) and Ag(1-y)Sb(y) phases, as well as the identical work functions, both thermal conductivity and electrical resistivity of the nanocomposites were significantly reduced in the lower temperature regime compared with pure FeSb(2). Overall, an improvement of 70% in ZT was achieved for the optimized nanocomposite FeSb(1.975)Ag(0.025)/Ag(0.77)Sb(0.23) sample, in which Ag(0.77)Sb(0.23) is about 10% by molar ratio. The results of this approach clearly demonstrated the metal/semiconductor interface concept and confirmed the potential of strongly correlated material systems as promising thermoelectric materials.

CoCrMo Metal-on-metal Hip Replacements

After the rapid growth in the use of CoCrMo metal-on-metal hip replacements since the second generation was introduced circa 1990, metal-on-metal hip replacements have experienced a sharp decline in the last two years due to biocompatibility issues related to wear and corrosion products. Despite some excellent clinical results, the release of wear and corrosion debris and the adverse response of local tissues have been of great concern. There are many unknowns regarding how CoCrMo metal bearings interact with the human body. This perspective article is intended to outline some recent progresses in understanding wear and corrosion of metal-on-metal hip replacement both in vivo and in vitro. The materials, mechanical deformation, corrosion, wear-assisted corrosion, and wear products will be discussed. Possible adverse health effects caused by wear products will be briefly addressed, as well as some of the many open questions such as the detailed chemistry of corrosion, tribochemical reactions and the formation of graphitic layers. Nowadays we design almost routinely for high performance materials and lubricants for automobiles; humans are at least as important. It is worth remembering that a hip implant is often the difference between walking and leading a relatively normal life, and a wheelchair.

Repeated Syncope in a Needle Man

Conceptualizing a Tool to Optimize Therapy Based on Dynamic Heterogeneity

Complex biological systems often display a randomness paralleled in processes studied in fundamental physics. This simple stochasticity emerges owing to the complexity of the system and underlies a fundamental aspect of biology called phenotypic stochasticity. Ongoing stochastic fluctuations in phenotype at the single-unit level can contribute to two emergent population phenotypes. Phenotypic stochasticity not only generates heterogeneity within a cell population, but also allows reversible transitions back and forth between multiple states. This phenotypic interconversion tends to restore a population to a previous composition after that population has been depleted of specific members. We call this tendency homeostatic heterogeneity. These concepts of dynamic heterogeneity can be applied to populations composed of molecules, cells, individuals, etc. Here we discuss the concept that phenotypic stochasticity both underlies the generation of heterogeneity within a cell population and can be used to control population composition, contributing, in particular, to both the ongoing emergence of drug resistance and an opportunity for depleting drug-resistant cells. Using notions of both 'large' and 'small' numbers of biomolecular components, we rationalize our use of Markov processes to model the generation and eradication of drug-resistant cells. Using these insights, we have developed a graphical tool, called a metronomogram, that we propose will allow us to optimize dosing frequencies and total course durations for clinical benefit.

Generalized Principles of Stochasticity Can Be Used to Control Dynamic Heterogeneity

It is increasingly appreciated that phenotypic stochasticity plays fundamental roles in biological systems at the cellular level and that a variety of mechanisms generates phenotypic interconversion over a broad range of time scales. The ensuing dynamic heterogeneity can be used to understand biological and clinical processes involving diverse phenotypes in different cell populations. The same principles can be applied, not only to populations composed of cells, but also to populations composed of molecules, tissues, and multicellular organisms. Stochastic units generating dynamic heterogeneity can be integrated across various length scales. We propose that a graphical tool we have developed, called a metronomogram, will allow us to identify factors that suitably influence the restoration of homeostatic heterogeneity so as to modulate the consequences of dynamic heterogeneity for desired outcomes.

[Effects of Mechanical Stimulation on Proliferation and Differentiation in MG-63 Osteoblast-like Cells]

This paper is aimed to explore the effects of mechanical stimulation on proliferation and differentiation of osteoblasts. Cultured MG-63 osteoblast-like cells were strained by the four-point bending cell mechanics loader. In the study, we observed the effects of different magnitudes and duration of mechanical strain on the markers of proliferation and differentiation in osteoblasts. The protein levels and Alkaline phosphatase (ALP) activity were determined by western blot and alpha-nitrophenyl phosphate assay respectively. The mineralization nodules were stained using Alizarin Red-S method. We found: (1) the expression of proliferating cell nuclear antigen (PCNA), ALP activity in strained group were significantly increased compared to those in the control group, but the role did not increase with the increase of the magnitude of the stimulation; and (2) under appropriate stimulation (2000 microstrain), the expression of PCNA, COL I protein and ALP activity increased gradually with the increase of loading time, and appropriate stimulation promoted the formation of mineralization nodules. It indicated that appropriate mechanical stimulation could promote proliferation and differentiation of osteoblasts.

[Optimization of Expression Conditions of Recombinant Fuantai-03 and Detection of Its Biological Activities]

Fuantai-03(FAT-03), isolated from the Dasyatis akajei, has a strong antiangiogenic activity. The recombinant Fuantai-03 (GST/rFAT-03) fusion protein can be obtained with the DNA recombination technology. In this study, expression conditions of GST/rFAT-03 were optimized by response surface experimental design method. The constructed engineering bacteria containing GST/rFAT-03 plasmid was induced by isopropy-beta-D-thiogalactosid (IPTG), the GST affinity column was used for isolation and purification, and then the effects of different culture time, IPTG concentration, induction temperature and induction time on the amount of soluble GST/rFAT-03 fusion protein were compared. The culture time for optimal expression was 6.13 h, IPTG concentration was 0.36 mmol/L, induction temperature was 19.71 degrees C, and induction time was 13.60 h. The amount of soluble GST/rFAT-03 fusion protein was 7.57 mg/L under above mentioned expression conditions. The results also showed that rFAT-03 significantly inhibited angiogenesis in chicken chorioallantoic membrane in a dose-dependent manner. Moreover, the soluble form of the target protein is useful for further work on purification and on studying its biological function.

How Comorbidities and Preoperative Expenditures Correlate with Postoperative Adverse Outcomes

Objectives: To examine the correlation of preexisting illnesses and preoperative medical expenditures with postoperative major adverse outcomes among geriatric surgical patients. Study Design: Retrospective cohort study using claims from Taiwan's National Health Insurance Research Database. Methods: All geriatric patients aged >65 years receiving inpatient surgeries during 2004 to 2007 under universal healthcare coverage were included. Surgical patients aged 55 to 64 years were the reference group. Risk-adjusted 30-day postoperative complication and mortality rates among elderly patients in various age sectors were analyzed and correlated with the preexisting illnesses and preoperative medical expenditures quantitatively. Results: Among 432,614 elderly surgical patients in specific age sectors and 238,802 controls, the prevalence of preexisting illnesses and the riskadjusted postoperative adverse outcome rates were highly age dependent and illness related. When comparing patients aged >85 years with patients aged 55 to 64 years, the adjusted odds ratios were 2.74 (95% confidence interval [CI], 2.67-2.82) and 3.56 (95% CI 3.31-3.84) for incidence of major postoperative complications and mortality after major complications, respectively. Numbers of preexisting illnesses increased in an age-dependent pattern and the preoperative 24-month medical expenditures increased incrementally with the numbers of comorbidities. Postoperative major complications, but not mortality rates, were highly correlated with the numbers of comorbidities and increased parallel with preoperative 24-month comorbidity-related medical expenditures, especially in the younger age group. Conclusions: Adjusting for preexisting covariates, geriatric patients had an age-dependent, illnessrelated, and expenditure-associated pattern of higher postoperative complication and mortality rates. The numbers of comorbidities and preoperative medical expenditures had high predictive value for postoperative adverse outcomes.

Resolvin D1 Attenuates Inflammation in Lipopolysaccharide-induced Acute Lung Injury Through a Process Involving the PPARgamma/NF-kappaB Pathway

ABSTRACT: BACKGROUND: Docosahexaenoic acid (DHA) and DHA-derived lipid mediators have recently been shown to possess anti-inflammatory and pro-resolving properties. In fact, DHA can down-regulate lipolysaccharide (LPS)-induced activation of NF-kappaB via a PPARgamma-dependent pathway. We sought to investigate the effects of the novel DHA-derived mediator resolvin D1 (RvD1) on LPS-induced acute lung injury and to determine whether these effects occur via a PPARgamma-dependent pathway. METHODS: BALB/c mice aged 6--8 weeks were randomly divided into seven groups: two control groups receiving saline or RvD1 (600 ng) without LPS; a control group receiving LPS only; an experimental group receiving RvD1 (300 ng) or RvD1 (600 ng), followed by LPS; a group receiving the PPARgamma antagonist GW9662; and a group receiving GW9662, then RvD1 (600 ng) and finally LPS. LPS (50 muM) and saline were administered intratracheally. RvD1 was injected intravenously 24 h and 30 min before LPS, while GW9662 was injected intravenously 30 min before RvD1. Mice were killed at 6, 12, and 24 h. Samples of bronchoalveolar lavage fluid (BALF) were analyzed for cell counts and cytokine analysis. Lung tissues were collected for histology, Western blotting and electrophoretic mobility shift assays (EMSAs). RESULTS: At all three time points, groups receiving either dose of RvD1 followed by LPS had significantly lower total leukocyte counts and levels of TNF-alpha and IL-6 levels in BALF than did the group given only LPS. RvD1 markedly attenuated LPS-induced lung inflammation at 24 h, based on hematoxylin-eosin staining of histology sections. RvD1 activated PPARgamma and suppressed IkappaBalpha degradation and NF-kappaB p65 nuclear translocation, based on Western blots and EMSAs. The PPARgamma inhibitor GW9662 partially reversed RvD1-induced suppression of IkappaBalpha degradation and p65 nuclear translocation. CONCLUSIONS: These results suggest that RvD1 may attenuate lung inflammation of LPS-induced acute lung injury by suppressing NF-kappaB activation through a mechanism partly dependent on PPARgamma activation.

Decompressed Percutaneous Vertebroplasty: A Secured Bone Cement Delivery Procedure for Vertebral Augmentation in Osteoporotic Compression Fractures

The purpose of this study was to assess the efficacy of a new assistive procedure for injecting cement in percutaneous vertebroplasty (PV). Percutaneous vertebroplasty is frequently used for treating patients with osteoporotic vertebral compression fractures. However, the leakage of bone cement during PV may lead to serious complications, such as spinal cord compression or pulmonary embolism. Herein we present a secure procedure designed to safely and effectively deliver the bone cement into the vertebral column. MATERIALS AND METHODS: Thirty-five patients with a total of 50 levels of osteoporotic compression fracture were consecutively recruited for the study. During a routine PV operation, acrylic cement was injected with a simultaneous application of a continuous negative pressure to the contralateral side of the vertebral body. This negative pressure exerts a pulling force that attracts the bone cement to flow within the vertebral body. RESULTS: With the proposed decompressed PV procedure, cross-filling of the vertebrographys was achieved for all 50 fracture levels, with no paravertebral venous plexus leakage. Three of the 50 levels (6%) exhibited contrast-medium leakage into the intradisc or cortical defect regions. After decompressed cement injection, excellent cross-filling of bone cement deposition was achieved in 38 of the 50 levels (76%; cement cross-filling region >75%), good cross-filling deposition was achieved in 7 levels (14%; cement cross-filling region >50%), deposition was poor in 3 levels (6%; cement cross-filling region <50%), and deposition failed in 2 levels (4%; fixed cement with no sign of cross-filling). Routine postoperative reviews revealed that six fracture levels (12%) had minimal cement leakage, with two leaking into the disc and four into paravertebral cortical defect regions. CONCLUSIONS: Compared to the reported 20-88% cement leakage rate for the conventional PV procedure, the proposed decompressed PV procedure offers a more secure and effective way to perform cement injection, and reduces the likelihood of cement leakage.

MicroRNA-137, a HMGA1 Target, Suppresses Colorectal Cancer Cell Invasion and Metastasis in Mice by Directly Targeting FMNL2

BACKGROUND & AIMS: Formin-like (FMNL)2 is upregulated in colorectal tumors and has been associated with tumor progression, but little is known about regulatory mechanisms. We investigated whether microRNAs (miRNAs) regulate levels of FMNL2 in colorectal cancer (CRC) cells. METHODS: We used real-time PCR and immunoblot analyses to measure levels of miR-137, high-mobility group AT-hook (HMGA)1, and FMNL2 in CRC cells and tissue samples from patients (n=50). We used luciferase reporter assays to determine the association between miR-137 and the FMNL2 3' untranslated region (UTR), and HMGA1 and the miR-137 promoter. Chromatin immunoprecipitation assays were used to assess direct binding of HMGA1 to the miR-137 promoter. RESULTS: miR-137 and miR-142-3p were predicted to bind FMNL2, based on bioinformatic data. Only the level of miR-137 had a significant inverse correlation with the level of FMNL2 protein in CRC cell lines and tissues. FMNL2 mRNA was targeted by miR-137; expression of miR-137 inhibited proliferation and invasion by CRC cells in vitro, and metastasis to liver and intestine by CRC xenografts in nude mice. HMGA1 bound to the promoter of miR-137 and activated its transcription, which reduced levels of FMNL2 in CRC cells. Ectopic expression of miR-137 in CRC cells inhibited phosphorylation of mitogen-activated protein kinase (MAPK) and Akt, which reduced levels of matrix metalloproteinase (MMP)2, MMP9, and vascular endothelial growth factor (VEGF); it also reduced invasiveness of CRC cells, inhibiting signaling via phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), Akt, and MAPK. CONCLUSIONS: Levels of miR-137 and HMGA1 are reduced, and levels of FMNL2, are increased in CRC samples compared with adjacent normal mucosa. In CRC cells, miR-137 targets FMNL2 mRNA and is regulated by the transcription factor HMGA1. Expression of miR-137 reduces CRC cell invasion in vitro and metastasis of tumor xenografts in mice. FMNL2 appears to activate PI3K, Akt, and MAPK signaling pathways.

Changes in Cell Autophagy and Apoptosis During Age-related Left Ventricular Remodeling in Mice and Their Potential Mechanisms

Cardiac structures and functions change with advanced age, but the underlying mechanisms are not well understood. Autophagy and apoptosis play important roles in the process of cardiac remodeling. This study was designed to explore changes in cell autophagy and apoptosis during age-related left ventricular remodeling and to determine whether the mitogen-activated protein kinase (MAPK) pathway is an underlying mechanism. Eight 5-month-old (adult group) and eight 24-month-old male C57bl/6 mice (aged group) were studied. The heart mass index, left ventricular mass index and hydroxyproline content of both groups were compared. Western blotting was used to quantify the protein expression of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, caspase-3, B-cell leukemia-2 (Bcl-2) and MAPKs in the left ventricles of adult and aged mice. Our results showed that the heart mass index, left ventricular mass index and hydroxyproline content in the left ventricles of the aged mice were increased significantly compared with the adult mice, indicating that left ventricular remodeling occurs with aging. The expression of LC3 and Beclin-1 in the left ventricles of aged mice were decreased significantly compared to adult mice. Meanwhile, the level of myocardial caspase-3 in adult mice remained the same in aged mice, and the level of myocardial Bcl-2 increased significantly in aged mice. There were no differences in the expression level of myocardial extracellular signal-regulated kinase 1/2 (ERK1/2), activated/phospho-ERK1/2, c-Jun N-terminal kinase 1/2 (JNK1/2) and p38 between aged and adult mice. However, the expression of myocardial activated/phospho-JNK1/2 increased significantly in aged mice, while activated/phospho-p38 decreased significantly. These findings indicate that autophagy decreases without a concurrent change in apoptosis during age-related left ventricular remodeling in mice. The MAPK pathway may be involved in the regulation of age-related left ventricular remodeling by modulating autophagy.

Phase Retrieval with Random Phase Illumination

This paper presents a detailed numerical study on the performance of the standard phasing algorithms with random phase illumination (RPI). Phasing with high resolution RPI and the oversampling ratio σ=4 determines a unique phasing solution up to a global phase factor. Under this condition, the standard phasing algorithms converge rapidly to the true solution without stagnation. Excellent approximation is achieved after a small number of iterations, not just with high resolution but also low resolution RPI in the presence of additive as well multiplicative noises. It is shown that RPI with σ=2 is sufficient for phasing complex-valued images under a sector condition and σ=1 for phasing nonnegative images. The error-reduction algorithm with RPI is proved to converge to the true solution under proper conditions.

Microbial Population Responses to PH and Salt Shock During Phenols Degradation Under High Salt Conditions Revealed by RISA and AFDRA

The responses of microbial community to pH and salt shock during phenols degradation under high salt conditions were revealed by two DNA fingerprint methods, i.e. ribosomal intergenic spacer analysis (RISA) and amplified functional DNA restriction analysis (AFDRA), together with 16S rDNA clone library analysis. It was shown that the phenols removal rate was improved with increasing NaCl concentration from 0 to 50 mg/L, and could remain at a high level even in the presence of 100 mg/L NaCl. The degradation efficiency remained stable under neutral conditions (pH 7.0-9.0), but decreased sharply under acidic (below pH 5.0) or more alkaline conditions (above pH 10.0). The community structure was dramatically changed during salt fluctuations, with Halomonas sp. and Marinobacter sp. as the predominant salt-tolerant species. Meanwhile, Marinobacter sp. and Alcaligenes faecalis sp. were the major species which might play the key role for stabilizing the treatment systems under different pH conditions. Moreover, the changes of phenol hydroxylase genes were analyzed by AFDRA, which showed that these functional genes were substantially different under any shock conditions.

[Functional MRI Analysis of Deception Among People with Antisocial Personality Disorders]

Objective: To investigate the functional magnetic resonance imaging (fMRI) data of deception in antisocial personality disorders (ASPD). Methods: A total of 32 criminals meeting the criteria for ASPD underwent fMRI at 1.5T while responding truthfully questions or lying. We compared the brain activities between truth-telling and lie-telling, and then computed the correlation coefficient between the contrast brain activities and the inclination to deception. Results: The left anterior cingulate gyrus, the bilateral dorsolateral prefrontal cortex, and left inferior parietal lobule were associated with the executive aspects of deception among people with ASPD. But with the greater inclination to deception, the blood oxygen level dependent (BOLD) activities in those regions decreased. Conclusion: Evaluations of truthful and untruthful communications pertaining to ASPD subjects may be differentiated in terms of brain BOLD activities, though those activities may decrease in habitual liars, which remains a challenge to the diagnostic accuracy in lie detection.

[1H-magnetic Resonance Spectroscopy on Bilateral Thalamus of Patients with Secondarily Generalized Tonic-clonic Seizures]

Objective: To examine the changes of metabolites in the bilateral thalamus of patients with secondarily generalized tonic-clonic seizure (SGTCS) and to explore the mechanism of SGTCS. Methods: Thirty patients with SGTCS (epilepsy group) and 30 matched healthy controls (control group) were examined by 1H-magnetic resonance spectroscopy (1H-MRS). The levels of N-acetyl aspartate (NAA), choline-containing compounds (Cho), creatine phosphocreatine (Cr-PCr), and myo-inositol (mI) of the bilateral thalamus were measured in both the epilepsy group and the control group. The ratios of NAA/Cr-PCr, NAA/(Cr-PCr+Cho), Cho/Cr-PCr and mI/Cr-PCr were compared and analyzed in the 2 groups. Results: The ratios of NAA/Cr-PCr, and NAA/(Cr-PCr+Cho)(1.7074 ± 0.2214; 0.9333 ± 0.2173) in the left thalamus in the epilepsy group were significantly lower than those in the control group(1.8834 ±0.2093; 1.1243 ±0.2447)(P<0.05). The ratios of NAA/Cr-PCr, and NAA/(Cr- PCr+Cho) (1.7472 ±0.2439; 0.9165 ±0.2462) in the right thalamus in the epilepsy group were also significantly lower than those in the control group(1.8925 ± 0.2004; 1.0941 ± 0.2372)(P<0.05). There were no significant differences in the ratios of NAA/Cr-PCr, NAA/(Cr-PCr+Cho), Cho/Cr- PCr, and mI/Cr-PCr between the bilateral thalamis in the epilepsy group (P>0.05). Conclusion: There is neuronal dysfunction in the bilateral thalamus in the epilepsy group. Abnormal changes of the bilateral thalamus are involved in the mechanism of SGTCS.

WIN55,212-2 Protects Oligodendrocyte Precursor Cells in Stroke Penumbra Following Permanent Focal Cerebral Ischemia in Rats

Aim:To explore whether the synthetic cannabinoid receptor agonist WIN55,212-2 could protect oligodendrocyte precursor cells (OPCs) in stroke penumbra, thereby providing neuroprotection following permanent focal cerebral ischemia in rats.Methods:Adult male SD rats were subjected to permanent middle cerebral artery occlusion (p-MCAO). The animals were administered WIN55,212-2 at 2 h, and sacrificed at 24 h after the ischemic insult. The infarct volumes and brain swelling were assessed. The expression of cannabinoid receptor type 1 (CB1) in the stroke penumbra was examined using Western blot assay. The pathological changes and proliferation of neural glial antigen 2-positive OPCs (NG2(+) cells) in the stroke penumbra were studied using immunohistochemistry staining.Results:p-MCAO significantly increased the expression of CB1 within the stroke penumbra with the highest level appearing at 2 h following the ischemic insult. Administration of WIN55,212-2 (9 mg/kg, iv) significantly attenuated the brain swelling, and reduced the infarct volume as well as the number of tau-immunoreactive NG2(+) cells (tau-1(+)/NG2(+) cells) in the stroke penumbra. Moreover, WIN55,212-2 significantly promoted the proliferation of NG2(+) cells in the stroke penumbra and in the ipsilateral subventricular zone at 24 h following the ischemic insult. Administration of the selective CB1 antagonist rimonabant (1 mg/kg, iv) partially blocked the effects caused by WIN55,212-2.Conclusion:Tau-1 is expressed in NG2(+) cells following permanent focal cerebral ischemic injury. Treatment with WIN55,212-2 reduces the number of tau-1(+)/NG2(+) cells and promotes NG2(+) cell proliferation in the stroke penumbra, which are mediated partially via CB1 and may contribute to its neuroprotective effects.

Sparse Patch-Based Label Propagation for Accurate Prostate Localization in CT Images

In this paper, we propose a new prostate CT segmentation method for image guided radiation therapy (IGRT). The main contributions of our method lie in the following aspects: (1) Instead of using voxel intensity information alone, patch-based representation in the discriminative feature space with logistic sparse LASSO is used as anatomical signature to deal with low contrast problem in prostate CT images. (2) Based on the proposed patch-based signature, a new multi-atlases label fusion method formulated under sparse representation framework is designed to segment prostate in the new treatment images, with guidance from the previous segmented images of the same patient. This method estimates the prostate likelihood of each voxel in the new treatment image from its nearby candidate voxels in the previous segmented images, based on the non-local mean principle and sparsity constraint. (3) A hierarchical labeling strategy is further designed to perform label fusion, where voxels with high confidence are first labeled for providing useful context information in the same image for aiding the labeling of the remaining voxels. (4) An online update mechanism is finally adopted to progressively collect more patient-specific information from newly segmented treatment images of the same patient, for adaptive and more accurate segmentation. The proposed method has been extensively evaluated on a prostate CT image database consisting of 24 patients where each patient has more than 10 treatment images, and further compared with several state-of-the-art prostate CT segmentation algorithms using various evaluation metrics. Experimental results demonstrate that the proposed method consistently achieves higher segmentation accuracy than any other methods under comparison.

Crystal-storing Histiocytosis in a Patient with Ocular Extranodal Marginal Zone Lymphoma

Implementation and Outcome of Thrombolysis with Alteplase 3 to 4.5 h After Acute Stroke in Chinese Patients

BACKGROUND AND PURPOSE: The European Cooperative Acute Stroke Study (ECASS) III showed that intravenous recombinant tissue plasminogen activator (rtPA) administered in the 3 to 4.5 h after symptom onset significantly improved clinical outcomes in patients with acute ischemic stroke (AIS). But little is known regarding the safety and efficacy of intravenous rtPA treatment within this extended time window in Chinese patients with AIS. METHODS AND RESULTS: Data were collected from the Thrombolysis Implementation and Monitor of acute ischemic Stroke in China (TIMS-China). A total of 574 patients who underwent rtPA therapy within 4.5 h after symptom onset were included in this study: 409 in the 0- to 3-h group and 165 in the 3- to 4.5-h group. There were no significant differences in SICH rate (2.4% vs. 1.5%, P = 0.70) at 24 to 36 h, mortalities (7.5% vs. 7.3%, P = 0.84), independence rate (68.9% vs. 63.9%, P = 0.19), and excellent recovery rate (60.9% vs. 52.4%, P = 0.11) between the two time window groups. These results were comparable with previous Western studies. CONCLUSION: This study suggests that intravenous rtPA treatment at 3 to 4.5 h of symptom onset remains safe and effective in Chinese patients with AIS.

Examining the Effects of an Outpatient Palliative Care Consultation on Symptom Burden, Depression, and Quality of Life in Patients with Symptomatic Heart Failure

We conducted this prospective comparative study to examine the feasibility and effectiveness of a palliative care consultation along with standard heart failure care in an outpatient setting regarding symptom burden, depression, and quality of life (QOL).

Characterization of in Vitro Modified Human Very Low-density Lipoprotein Particles and Phospholipids by Capillary Electrophoresis

A simple capillary zone electrophoresis (CZE) method was used to characterize human very low-density lipoprotein (VLDL) particles for four healthy donors. One major peak was observed for native, in vitro oxidized and glycated VLDL particles. The effective mobilities and peak areas of the capillary electrophoresis (CE) profiles showed good reproducibility and precision. The mobility of the oxidized VLDL peak was higher than that of the native VLDL. The mobility of the glycated VLDL peak was similar to that of the native VLDL. Phospholipids isolated from VLDL particles were analyzed by our recently developed micellar electrokinetic chromatography (MEKC) with a high-salt stacking method. At absorbance 200 nm, the native VLDL phospholipids showed a major peak and a minor peak for each donor. For oxidized VLDL phospholipids, the area of the major peak reduced for three donors, possibly due to phospholipid decomposition. For glycated VLDL phospholipids, the peak mobilities were more positive than native VLDL phospholipids for two donors, possibly due to phospholipid-linked advanced glycation end products (AGEs). Very interestingly, at absorbance 234 nm, the major peak of oxidized VLDL phospholipids was resolved as two peaks for each donor, possibly due to conjugated dienes formed upon oxidation.

Fetal Liver Bisphenol A Concentrations and Biotransformation Gene Expression Reveal Variable Exposure and Altered Capacity for Metabolism in Humans

Widespread exposure to the endocrine active compound, bisphenol A (BPA), is well documented in humans. A growing body of literature suggests adverse health outcomes associated with varying ranges of exposure to BPA. In the current study, we measured the internal dose of free BPA and conjugated BPA and evaluated gene expression of biotransformation enzymes specific for BPA metabolism in 50 first- and second-trimester human fetal liver samples. Both free BPA and conjugated BPA concentrations varied widely, with free BPA exhibiting three times higher concentrations than conjugated BPA concentrations. As compared to gender-matched adult liver controls, UDP-glucuronyltransferase, sulfotransferase, and steroid sulfatase genes exhibited reduced expression whereas β-glucuronidase mRNA expression remained unchanged in the fetal tissues. This study provides evidence that there is considerable exposure to BPA during human pregnancy and that the capacity for BPA metabolism is altered in the human fetal liver.

Lymphatic Vessels in Health and Disease

The lymphatic vasculature plays vital roles in tissue fluid balance, immune defense, metabolism, and cancer metastasis. In adults, lymphatic vessel formation and remodeling occur primarily during inflammation, development of the corpus luteum, wound healing, and tumor growth. Unlike the blood circulation, where unidirectional flow is sustained by the pumping actions of the heart, pumping actions intrinsic to the lymphatic vessels themselves are important drivers of lymphatic flow. This review summarizes critical components that control lymphatic physiology. WIREs Syst Biol Med 2012. doi: 10.1002/wsbm.1201 For further resources related to this article, please visit the WIREs website.

CD11b/CD18 (Mac-1) Is a Novel Surface Receptor for Extracellular Double-Stranded RNA To Mediate Cellular Inflammatory Responses

During viral infection, extracellular dsRNA is a potent signaling molecule that activates many innate immune cells, including macrophages. TLR3 is a well-known receptor for extracellular dsRNA, and internalization of extracellular dsRNA is required for endosomal TLR3 activation. Preserved inflammatory responses of TLR3-deficient macrophages to extracellular dsRNA strongly support a TLR3-independent mechanism in dsRNA-mediated immune responses. The present study demonstrated that CD11b/CD18 (Mac-1 [macrophage-1 Ag]), a surface integrin receptor, recognized extracellular dsRNA and induced macrophage immune responses. CD11b deficiency reduced inflammatory cytokine induction elicited by polyinosinic:polycytidylic acid (poly I:C; a synthetic dsRNA) in mouse sera and livers, as well as in cultured peritoneal macrophages. dsRNA-binding assay and confocal immunofluorescence showed that Mac-1, especially the CD11b subunit, interacted and colocalized with poly I:C on the surface of macrophages. Further mechanistic studies revealed two distinct signaling events following dsRNA recognition by Mac-1. First, Mac-1 facilitated poly I:C internalization through the activation of PI3K signaling and enhanced TLR3-dependent activation of IRF3 in macrophages. Second, poly I:C induced activation of phagocyte NADPH oxidase in a TLR3-independent, but Mac-1-dependent, manner. Subsequently, phagocyte NADPH oxidase-derived intracellular reactive oxygen species activated MAPK and NF-κB pathways. Our results indicate that extracellular dsRNA activates Mac-1 to enhance TLR3-dependent signaling and to trigger TLR3-independent, but Mac-1-dependent, inflammatory oxidative signaling, identifying a novel mechanistic basis for macrophages to recognize extracellular dsRNA to regulate innate immune responses. This study identifies Mac-1 as a novel surface receptor for extracellular dsRNA and implicates it as a potential therapeutic target for virus-related inflammatory diseases.

The Principal Genetic Determinants for Nasopharyngeal Carcinoma in China Involve the HLA Class I Antigen Recognition Groove

Nasopharyngeal carcinoma (NPC) is an epithelial malignancy facilitated by Epstein-Barr Virus infection. Here we resolve the major genetic influences for NPC incidence using a genome-wide association study (GWAS), independent cohort replication, and high-resolution molecular HLA class I gene typing including 4,055 study participants from the Guangxi Zhuang Autonomous Region and Guangdong province of southern China. We detect and replicate strong association signals involving SNPs, HLA alleles, and amino acid (aa) variants across the major histocompatibility complex-HLA-A, HLA -B, and HLA -C class I genes (P(HLA-A-aa-site-62) = 7.4×10(-29); P (HLA-B-aa-site-116) = 6.5×10(-19); P (HLA-C-aa-site-156) = 6.8×10(-8) respectively). Over 250 NPC-HLA associated variants within HLA were analyzed in concert to resolve separate and largely independent HLA-A, -B, and -C gene influences. Multivariate logistical regression analysis collapsed significant associations in adjacent genes spanning 500 kb (OR2H1, GABBR1, HLA-F, and HCG9) as proxies for peptide binding motifs carried by HLA- A*11:01. A similar analysis resolved an independent association signal driven by HLA-B*13:01, B*38:02, and B*55:02 alleles together. NPC resistance alleles carrying the strongly associated amino acid variants implicate specific class I peptide recognition motifs in HLA-A and -B peptide binding groove as conferring strong genetic influence on the development of NPC in China.

Construction of High-Density Genetic Linkage Maps and Mapping of Growth-Related Quantitative Trail Loci in the Japanese Flounder (Paralichthys Olivaceus)

High-density genetic linkage maps were constructed for the Japanese flounder (Paralichthys olivaceus). A total of 1624 microsatellite markers were polymorphic in the reference family. Linkage analysis using JoinMap 4.0 resulted in the mapping of 1487 markers to 24 linkage groups, a result which was consistent with the 24 chromosomes seen in chromosome spreads. The female map was composed of 1257 markers, covering a total of 1663.8 cM with an average interval 1.35 cM between markers. The male map consisted of 1224 markers, spanning 1726.5 cM, with an average interval of 1.44 cM. The genome length in the Japanese flounder was estimated to be 1730.3 cM for the females and 1798.0 cM for the males, a coverage of 96.2% for the female and 96.0% for the male map. The mean recombination at common intervals throughout the genome revealed a slight difference between sexes, i.e. 1.07 times higher in the male than female. High-density genetic linkage maps are very useful for marker-assisted selection (MAS) programs for economically valuable traits in this species and for further evolutionary studies in flatfish and vertebrate species. Furthermore, four quantiative trait loci (QTL) associated with growth traits were mapped on the genetic map. One QTL was identified for body weight on LG 14 f, which explained 14.85% of the total variation of the body weight. Three QTL were identified for body width on LG14f and LG14m, accounting for 16.75%, 13.62% and 13.65% of the total variation in body width, respectively. The additive effects were evident as negative values. There were four QTL for growth traits clustered on LG14, which should prove to be very useful for improving growth traits using molecular MAS.

Expansion of Hematopoietic Stem Cells for Transplantation: Current Perspectives

ABSTRACT: Hematopoietic stem cells (HSCs) are rare cells that have the unique ability to self-renew and differentiate into cells of all hematopoietic lineages. The expansion of HSCs has remained an important goal to develop advanced cell therapies for bone marrow transplantation and many blood disorders. Over the last several decades, there have been numerous attempts to expand HSCs in vitro using purified growth factors that are known to regulate HSCs. However, these attempts have been met with limited success for clinical applications. New developments in the HSC expansion field coupled with gene therapy and stem cell transplant should encourage progression in attractive treatment options for many disorders including hematologic conditions, immunodeficiencies, and genetic disorders.

Numerical Analysis of Airflow Alteration in Central Airways Following Tracheobronchial Stent Placement

ABSTRACT: The computational fluid dynamics method, which provides an estimation of the pressure drop in the airway before and after the stent implantation, is proposed in this study. This method is based on the finite volume model. The pressure field was solved by the Navier-Stokes equations. The proposed methodology was evaluated in seven health people (control group) and in fourteen patients who were assigned in two groups, in which one was tracheal stenosis and the other was bronchial stenosis. The results showed that the pressure drop after tracheal stent implantation became significantly smaller. For bronchial stent implantation cases, the airway resistance improved insignificantly.

Steroid Receptor Coactivator-1 Mediates Estrogenic Actions to Prevent Body Weight Gain in Female Mice

Estrogen receptor-α (ERα) expressed by hypothalamic proopiomelanocortin and steroidogenic factor-1 neurons largely mediates the antiobesity effects of estrogens in females. However, the critical molecular events that are coupled to ERα and mediate estrogenic effects on energy balance remain unknown. In the current study, we demonstrated that steroid receptor coactivator-1 (SRC1), a nuclear receptor coactivator, is abundantly expressed by both proopiomelanocortin and steroidogenic factor-1 neurons. We further showed that central administration of an ERα agonist, propyl pyrazole triol, acutely increases physical interaction between SRC1 and ERα in the hypothalamus. Finally, we demonstrated that the effects of estrogens on energy homeostasis are significantly blunted in female mice lacking SRC1 globally. Collectively our results indicate that SRC1 is functionally required to mediate the antiobesity effects of estrogen-ERα signals.

Continuous Infusion of N-acetylcysteine by Nasobiliary for Advanced Intraductal Papillary Mucinous Neoplasm of Bile Ducts (With Video)

Adherence to National Guidelines for Antiemesis Prophylaxis in Patients Undergoing Chemotherapy for Lung Cancer: A Population-based Study

BACKGROUND: Nausea and vomiting (N/V) during chemotherapy can have profound clinical and economic consequences. Effective antiemetic agents are available for prophylaxis, but barriers may prevent their use. For this population-based study, the authors assessed the rates of antiemetic prophylaxis use, and predictors of such use, among patients who were receiving platinum-based chemotherapy for lung cancer between 2001 and 2007. METHODS: The authors searched the Texas Cancer Registry-Medicare-linked database for individuals aged >65 years who received platinum-based chemotherapy within 12 months after a first diagnosis of lung cancer from 2001 to 2007; and all patients had continuous Medicare Part A and Part B coverage for the same period. Adherence to recommended regimens for N/V prophylaxis (established by the National Comprehensive Cancer Network) was scored as a binary variable (adherent vs nonadherent) and was calculated as the percentages of treated patients receiving each recommended agent within 1 day of beginning chemotherapy. Logistic regression with stepwise selection was used to examine whether patient characteristics influenced adherence. RESULTS: Of 4566 selected patients, adherence rates for the receipt of serotonin antagonists (eg, ondansetron) with dexamethasone were 60% to 90% regardless of whether the chemotherapy agent was considered moderately or highly emetogenic. The receipt of substance-P antagonists was much less common (<10%) during any period. On multivariate logistic regression modeling, variables that predicted adherence were older age, white race, higher median income, and concurrent radiation therapy. CONCLUSIONS: Recommended use of antiemetics for prophylaxis, especially substance-P antagonists, during chemotherapy for lung cancer is suboptimal. Factors that were correlated with adherence suggest socioeconomic barriers in the community. Cancer 2012. © 2012 American Cancer Society.

Characterization of Neuroblastic Tumors Using 18F-FDOPA PET

Neuroblastic tumors are childhood neoplasms that possess amino acid decarboxylase (AADC) activity and can theoretically be imaged by (18)F-fluorodihydroxyphenylalanine ((18)F-FDOPA) PET, a new diagnostic tool for neuroendocrine tumors. In this study, we explored the accuracy and clinical role of (18)F-FDOPA PET in neuroblastic tumors. METHODS: From 2008 to 2011, patients with tissue-proven neuroblastic tumors receiving (18)F-FDOPA PET at initial diagnosis or during follow-ups were enrolled. The sensitivity and specificity of (18)F-FDOPA PET were compared with those of (123)I-metaiodobenzylguanidine ((123)I-MIBG) scintigraphy and (18)F-FDG PET, using tumor histology as the standard. The maximum standardized uptake value and tumor-to-liver uptake ratio on (18)F-FDOPA PET were measured and correlated with AADC messenger RNA level in tumor tissue. RESULTS: Fifty tumors from 34 patients, including 42 neuroblastic tumors and 8 lesions without viable tumor cells, were eligible for analysis. (18)F-FDOPA PET successfully detected neuroblastic tumors of different histologic types in various anatomic sites, at a sensitivity of 97.6% (87.4%-99.9%) and a specificity of 87.5% (47.3%-99.7%). In tumors with concomitant studies, (18)F-FDOPA PET demonstrated a higher sensitivity than (123)I-MIBG scintigraphy (n = 18; P = 0.0455) or (18)F-FDG PET (n = 46; P = 0.0455). Among the 18 tumors with concomitant (123)I-MIBG scans, 4 tumors with viable cells were (123)I-MIBG-negative but were successfully detected by (18)F-FDOPA PET. The tumor uptake of (18)F-FDOPA significantly correlated with AADC expression (n = 15 nonhepatic tumors; maximum standardized uptake value, P = 0.0002; tumor-to-liver uptake ratio, P < 0.0001). CONCLUSION: (18)F-FDOPA PET showed high sensitivity and specificity in detecting and tracking neuroblastic tumors in this preliminary study with a small cohort of patients and might be complementary to (123)I-MIBG scintigraphy and (18)F-FDG PET. By correlating with AADC expression, (18)F-FDOPA PET might serve as a useful imaging tool for the functional assessment of neuroblastic tumors.

ChIP for Identification of P53 Responsive DNA Promoters

Chromatin immunoprecipitation assay (ChIP) has been frequently used to determine whether a transcriptional regulator can bind to a specific DNA element in the chromatin content of cells. Here, we describe a detailed protocol for this assay with hands-on tips based on our own experience in working on the transcriptional regulator and tumor suppressor p53.

Features of Airway Remodeling in Different Types of Chinese Chronic Rhinosinusitis Are Associated with Inflammation Patterns

The remodeling patterns in different types of chronic rhinosinusitis (CRS) have rarely been compared, particularly the difference between eosinophilic and noneosinophilic CRS with nasal polyps (CRSwNP). Moreover, whether there is a link between remodeling and inflammation remains controversial.

Electrical Coupling of Isolated Cardiomyocyte Clusters Grown on Aligned Conductive Nanofibrous Meshes for Their Synchronized Beating

Myocardial infarction is often associated with abnormalities in electrical function due to a massive loss of functioning cardiomyocytes. This work develops a mesh, consisting of aligned composite nanofibers of polyaniline (PANI) and poly(lactic-co-glycolic acid) (PLGA), as an electrically active scaffold for coordinating the beatings of the cultured cardiomyocytes synchronously. Following doping by HCl, the electrospun fibers could be transformed into a conductive form carrying positive charges, which could then attract negatively charged adhesive proteins (i.e. fibronectin and laminin) and enhance cell adhesion. During incubation, the adhered cardiomyocytes became associated with each other and formed isolated cell clusters; the cells within each cluster elongated and aligned their morphology along the major axis of the fibrous mesh. After culture, expression of the gap-junction protein connexin 43 was clearly observed intercellularly in isolated clusters. All of the cardiomyocytes within each cluster beat synchronously, implying that the coupling between the cells was fully developed. Additionally, the beating rates among these isolated cell clusters could be synchronized via an electrical stimulation designed to imitate that generated in a native heart. Importantly, improving the impaired heart function depends on electrical coupling between the engrafted cells and the host myocardium to ensure their synchronized beating.

Ethyl Acetate Extraction from a Chinese Herbal Formula, Jiedu Xiaozheng Yin, Inhibits the Proliferation of Hepatocellular Carcinoma Cells Via Induction of G0/G1 Phase Arrest In vivo and In vitro

Jiedu Xiaozheng Yin (JXY), a polyherbal formula of traditional Chinese medicine (TCM), has been used to treat various kinds of cancer in China. However, the mechanism of its anticancer activity has yet to be elucidated. Air-dried herbs were extracted with reagents of different polarity. HepG2 cells were treated with different doses of ethyl acetate extract (EE-JXY) and chloroform extract (CE-JXY) for 24 h. Cell viability was detected by MTT assay. Colony formation ability was also evaluated. Cell cycle was evaluated by FACS. Tumor bearing BALB/c nude mice was treated with EE-JXY (0.06 g/kg) for 20 days. Tumor volume and weight were monitored. The percentage of PCNA-positive cells and the level of G1 phase proteins [cyclin-dependent kinase2 (CDK2), cyclin‑dependent kinase4 (CDK4), cyclin D and cyclin E and G2 phase proteins [cyclin-dependent kinase1 (CDK1), cyclin A and cyclin B] were detected by immunohistochemistry and western blotting. EE-JXY and CE-JXY dose-dependently inhibited the growth of HepG2 cells (P<0.01 for both). Furthermore, EE-JXY inhibited the formation of cell colonies and blocked the cell cycle to G1 phase in a dose-dependent manner (P<0.01 for all). EE-JXY showed an obviously antitumor effect in vivo (P<0.05). Further investigation showed that EE-JXY decreased the proliferation index of tumors (P<0.01) through increasing the expression of G1-related proteins (cyclin D and cyclin E, P<0.05 and P<0.01). These results suggested that JXY inhibits the growth of HepG2 cells at least via arresting the cell cycle at the G0/G1 phase.

Letter: Pyogenic Liver Abscess and Colorectal Cancer

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