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1.  Kv2 dysfunction after peripheral axotomy enhances sensory neuron responsiveness to sustained input☆ 
Experimental Neurology  2014;251(100):115-126.
Peripheral nerve injuries caused by trauma are associated with increased sensory neuron excitability and debilitating chronic pain symptoms. Axotomy-induced alterations in the function of ion channels are thought to largely underlie the pathophysiology of these phenotypes. Here, we characterise the mRNA distribution of Kv2 family members in rat dorsal root ganglia (DRG) and describe a link between Kv2 function and modulation of sensory neuron excitability. Kv2.1 and Kv2.2 were amply expressed in cells of all sizes, being particularly abundant in medium-large neurons also immunoreactive for neurofilament-200. Peripheral axotomy led to a rapid, robust and long-lasting transcriptional Kv2 downregulation in the DRG, correlated with the onset of mechanical and thermal hypersensitivity. The consequences of Kv2 loss-of-function were subsequently investigated in myelinated neurons using intracellular recordings on ex vivo DRG preparations. In naïve neurons, pharmacological Kv2.1/Kv2.2 inhibition by stromatoxin-1 (ScTx) resulted in shortening of action potential (AP) after-hyperpolarization (AHP). In contrast, ScTx application on axotomized neurons did not alter AHP duration, consistent with the injury-induced Kv2 downregulation. In accordance with a shortened AHP, ScTx treatment also reduced the refractory period and improved AP conduction to the cell soma during high frequency stimulation. These results suggest that Kv2 downregulation following traumatic nerve lesion facilitates greater fidelity of repetitive firing during prolonged input and thus normal Kv2 function is postulated to limit neuronal excitability. In summary, we have profiled Kv2 expression in sensory neurons and provide evidence for the contribution of Kv2 dysfunction in the generation of hyperexcitable phenotypes encountered in chronic pain states.
Highlights
•Kv2.1 and Kv2.2 are expressed in rat dorsal root ganglion neurons.•Kv2 subunits are most abundant in myelinated sensory neurons.•Kv2.1 and Kv.2 subunits are downregulated in a traumatic nerve injury pain model.•Kv2 inhibition ex vivo allows higher firing rates during sustained stimulation.•We conclude that Kv2 channels contribute to limiting peripheral neuron excitability.
doi:10.1016/j.expneurol.2013.11.011
PMCID: PMC3898477  PMID: 24252178
AP, action potential; APD50, AP half width; AHPD50, after-hyperpolarization half width; ATF3, activating transcription factor 3; CGRP, calcitonin gene-related peptide; CNS, central nervous system; DRG, dorsal root ganglion; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IB4, isolectin B4; IHC, immunohistochemistry; IR, input resistance; ISH, in situ hybridization; Kv channel, voltage-gated potassium channel; NF200, neurofilament 200; RP, refractory period; ScTx, stromatoxin-1; SNT, spinal nerve transection; Neuropathic pain; Potassium channel; Dorsal root ganglia
2.  Downstream Signaling Pathways in Mouse Adipose Tissues Following Acute In Vivo Administration of Fibroblast Growth Factor 21 
PLoS ONE  2013;8(9):e73011.
FGF21 is a novel secreted protein with robust anti-diabetic, anti-obesity, and anti-atherogenic activities in preclinical species. In the current study, we investigated the signal transduction pathways downstream of FGF21 following acute administration of the growth factor to mice. Focusing on adipose tissues, we identified FGF21-mediated downstream signaling events and target engagement biomarkers. Specifically, RNA profiling of adipose tissues and phosphoproteomic profiling of adipocytes, following FGF21 treatment revealed several specific changes in gene expression and post-translational modifications, specifically phosphorylation, in several relevant proteins. Affymetrix microarray analysis of white adipose tissues isolated from both C57BL/6 (fed either regular chow or HFD) and db/db mice identified over 150 robust potential RNA transcripts and over 50 potential secreted proteins that were changed greater than 1.5 fold by FGF21 acutely. Phosphoprofiling analysis identified over 130 phosphoproteins that were modulated greater than 1.5 fold by FGF21 in 3T3-L1 adipocytes. Bioinformatic analysis of the combined gene and phosphoprotein profiling data identified a number of known metabolic pathways such as glucose uptake, insulin receptor signaling, Erk/Mapk signaling cascades, and lipid metabolism. Moreover, a number of novel events with hitherto unknown links to FGF21 signaling were observed at both the transcription and protein phosphorylation levels following treatment. We conclude that such a combined "omics" approach can be used not only to identify robust biomarkers for novel therapeutics but can also enhance our understanding of downstream signaling pathways; in the example presented here, novel FGF21-mediated signaling events in adipose tissue have been revealed that warrant further investigation.
doi:10.1371/journal.pone.0073011
PMCID: PMC3765203  PMID: 24039848
3.  Ziyuglycoside II Inhibits the Growth of Human Breast Carcinoma MDA-MB-435 Cells via Cell Cycle Arrest and Induction of Apoptosis through the Mitochondria Dependent Pathway 
Ziyuglycoside II is one of the major active compounds of Sanguisorba officinalis L., which has a wide range of clinical applications including hemostasis, antibiosis, anti-inflammation and anti-oxidation. This study investigated the effect of ziyuglycoside II on the growth of human breast carcinoma MDA-MB-435 cells for the first time. The results showed that ziyuglycoside II could significantly inhibit the growth of MDA-MB-435 cells through blocking cell cycle progression at G0/G1 and S phase as well as via inducing cell apoptosis. Accumulation of reactive oxygen species (ROS) was observed in the progression of cell cycle arrest, which was associated with the increased expression of cell cycle regulating factors, p53 and p21. Subsequent apoptosis induced by ziyuglycoside II was accompanied with the activation of mitochondrial pathway, in particular a decreased mitochondrial membrane potential (MMP) as well as increased Bax/Bcl-2 ratio, cytochrome c release and the activity of caspase-3 and caspase-9. In conclusion, our study was the first to report that ziyuglycoside II has inhibitory effect on the growth of MDA-MB-435 cells, which might become a potential therapeutic approach of breast cancer in the future.
doi:10.3390/ijms140918041
PMCID: PMC3794768  PMID: 24005866
ziyuglycoside II; MDA-MB-435; cell cycle arrest; cell apoptosis
4.  Sensory Neuron Downregulation of the Kv9.1 Potassium Channel Subunit Mediates Neuropathic Pain following Nerve Injury 
Chronic neuropathic pain affects millions of individuals worldwide, is typically long-lasting, and remains poorly treated with existing therapies. Neuropathic pain arising from peripheral nerve lesions is known to be dependent on the emergence of spontaneous and evoked hyperexcitability in damaged nerves. Here, we report that the potassium channel subunit Kv9.1 is expressed in myelinated sensory neurons, but is absent from small unmyelinated neurons. Kv9.1 expression was strongly and rapidly downregulated following axotomy, with a time course that matches the development of spontaneous activity and pain hypersensitivity in animal models. Interestingly, siRNA-mediated knock-down of Kv9.1 in naive rats led to neuropathic pain behaviors. Diminished Kv9.1 function also augmented myelinated sensory neuron excitability, manifested as spontaneous firing, hyper-responsiveness to stimulation, and persistent after-discharge. Intracellular recordings from ex vivo dorsal root ganglion preparations revealed that Kv9.1 knock-down was linked to lowered firing thresholds and increased firing rates under physiologically relevant conditions of extracellular potassium accumulation during prolonged activity. Similar neurophysiological changes were detected in animals subjected to traumatic nerve injury and provide an explanation for neuropathic pain symptoms, including poorly understood conditions such as hyperpathia and paresthesias. In summary, our results demonstrate that Kv9.1 dysfunction leads to spontaneous and evoked neuronal hyperexcitability in myelinated fibers, coupled with development of neuropathic pain behaviors.
doi:10.1523/JNEUROSCI.3561-12.2012
PMCID: PMC3713313  PMID: 23197740
5.  Adenovirus-mediated delivery of interferon-γ gene inhibits the growth of nasopharyngeal carcinoma 
Background
Interferon-γ (IFN-γ) is regarded as a potent antitumor agent, but its clinical application is limited by its short half-life and significant side effects. In this paper, we tried to develop IFN-γ gene therapy by a replication defective adenovirus encoding the human IFN-γ (Ad-IFNγ), and evaluate the antitumoral effects of Ad-IFNγ on nasopharyngeal carcinoma (NPC) cell lines in vitro and in xenografts model.
Methods
The mRNA levels of human IFN-γ in Ad-IFNγ-infected NPC cells were detected by reverse transcription-polymerase chain reaction (RT-PCR), and IFN-γ protein concentrations were measured by enzyme-linked immunosorbent assay (ELISA) in the culture supernatants of NPC cells and tumor tissues and bloods of nude mice treated with Ad-IFNγ. The effects of Ad-IFNγ on NPC cell proliferation was determined using MTT assay, cell cycle distribution was determined by flow cytometry analysis for DNA content, and cells apoptosis were analyzed by Annexin V-FITC/7-AAD binding assay and hoechst 33342/PI double staining. The anti-tumor effects and toxicity of Ad-IFNγ were evaluated in BALB/c nude mice carrying NPC xenografts.
Results
The results demonstrated that Ad-IFNγ efficiently expressed human IFN-γ protein in NPC cell lines in vitro and in vivo. Ad-IFNγ infection resulted in antiproliferative effects on NPC cells by inducing G1 phase arrest and cell apoptosis. Intratumoral administration of Ad-IFNγ significantly inhibited the growth of CNE-2 and C666-1 cell xenografts in nude mice, while no significant toxicity was observed.
Conclusions
These findings indicate IFN-γ gene therapy mediated by replication defective adenoviral vector is likely a promising approach in the treatment of nasopharyngeal carcinoma.
doi:10.1186/1479-5876-10-256
PMCID: PMC3573957  PMID: 23272637
Gene therapy; Interferon-γ; Nasopharyngeal carcinoma; Adenoviral vector
6.  Body mass index, recreational physical activity and female urinary incontinence in Gansu, China 
Objective
To elucidate the influence of recreational physical activity, body mass index (BMI), and waist circumference on the risk of specific types of urinary incontinence.
Study design
We conducted a population-based cross-sectional survey in Gansu, China among 2603 women aged 20 years or older.
Results
The study found that BMI was positively associated with urinary incontinence (P for trend = 0.008) and the association was mainly observed for stress urinary incontinence (OR = 1.4, 95% CI: 1.1, 1.9 for BMI = 24.0–27.9 kg/m2; OR = 2.3, 95% CI: 1.5, 3.6 for BMI ≥ 28.0 kg/m2; P for trend = 0.0005). A positive association between stress incontinence (OR = 1.7, 95% CI: 1.2, 2.5) and waist circumference was observed for women who had waist circumference between 70 cm and 75 cm compared to waist circumference less than 70 cm. Recreational physical activity was inversely associated with overall and mixed urinary incontinence (P for trend <0.0001 for both). A significant interaction between physical activity and waist circumference was found for overall (P = 0.0007) and stress incontinence (P = 0.001).
Conclusions
The findings that physical activity inversely associated with urinary incontinence and its interaction with waist circumference warrant further investigation, particularly in prospective studies.
doi:10.1016/j.ejogrb.2011.07.016
PMCID: PMC3229870  PMID: 21821342
Recreational physical activity; Body mass index; Waist circumference; Female urinary incontinence
7.  Preliminary study of an intestinal bio-robot system based on nerve stimulation 
Background
Wireless capsule endoscopes for diagnosis and treatment in the gastrointestinal tract face the common problem of active actuation. To tackle this difficulty, a non-invasive intestinal bio-robot system with active actuation based on nerve stimulation was developed.
Methods
This intestinal inspection system utilized a natural organism—the mud eel—to serve as the locomotion mechanism, and it was controlled by a LabVIEW-programmed pulse generator. The exterior control unit was able to actively drive and remotely control the navigation and site-specific anchoring of the organism.
Results
Through in vitro stimulation experiments, a method of controlling the organism’s forward motion was obtained: when the organism was stimulated at the tail, it moved forward at a relatively fast speed and with high repeatability. The stimulator parameters were as follows: amplitude 1.85 μA, frequency 2 Hz, pulse duration 500 μs.
Conclusions
Since this is a preliminary study, considerable work remains to be done. However, the results could provide a solid theoretical basis for further research toward producing a practical intestinal bio-robot for the diagnosis and treatment of the gastrointestinal tract.
doi:10.1186/1743-0003-9-68
PMCID: PMC3546848  PMID: 23020999
Intestinal bio-robot system; Intestinal examination; Nerve stimulation; Locomotion control
8.  Overexpression of Fc receptor-like 1 associated with B-cell activation during hepatitis B virus infection 
The role of B cells in the pathogenesis of hepatitis B virus (HBV) infection has not been explored in depth. In the present study, the activation status of B cells from peripheral blood of healthy controls (N = 20) and patients with acute hepatitis B (AHB, N = 15) or chronic hepatitis B (CHB, N = 30) was evaluated by measuring the expression levels of B-cell activation markers CD69 and CD86, using quantitative real-time PCR and flow cytometry. Moreover, the potential mechanism underlying B-cell activation during HBV infection was further investigated by analyzing the expression profile of FCRL1, an intrinsic activation molecule of B cells. An elevation in the levels of B-cell activation markers including CD69 and CD86 was observed in the AHB patients (44.31 ± 9.27, 27.64 ± 9.26%) compared to CHB patients (30.35 ± 11.27, 18.41 ± 6.56%, P < 0.05), which was still higher than healthy controls (12.23 ± 7.84, 8.22 ± 3.43%, P < 0.05). Furthermore, the expression of FCRL1 was found to be similar to B-cell activation markers, which was highest in AHB patients (70.15 ± 17.11%), lowest in healthy donors (36.32 ± 9.98%, P < 0.05) and half-way between these levels in patients with CHB (55.17 ± 12.03%, P < 0.05). The results were positively associated with aberrant B-cell activation. These data suggest that B cells can play a role in HBV infection, and therefore more effort should be devoted to exploring their functions.
doi:10.1590/S0100-879X2012007500130
PMCID: PMC3854230  PMID: 22892829
Hepatitis B virus; B-cell activation; Fc receptor-like 1; Gene expression
9.  Canine Hip Dysplasia is Predictable by Genotyping 
Summary
Objective
To establish a predictive method using whole genome genotyping for early intervention in canine hip dysplasia (CHD) risk management, for the prevention of the progression of secondary osteoarthritis (OA), and for selective breeding.
Design
Two sets of dogs (6 breeds) were genotyped with dense SNPs covering the entire canine genome. The first set contained 359 dogs upon which a predictive formula for genomic breeding value (GBV) was derived by using their estimated breeding value (EBV) of the Norberg angle (a measure of CHD) and their genotypes. To investigate how well the formula would work for an individual dog with genotype only (without using EBV or phenotype), a cross validation was performed by masking the EBV of one dog at a time. The genomic data and the EBV of the remaining dogs were used to predict the GBV for the single dog that was left out. The second set of dogs included 38 new Labrador retriever dogs, which had no pedigree relationship to the dogs in the first set.
Results
The cross validation showed a strong correlation (r>0.7) between the EBV and the GBV. The independent validation showed a strong correlation (r=0.5) between GBV for the Norberg angle and the observed Norberg angle (no EBV was available for the new 38 dogs). Sensitivity, specificity, positive, and negative predictive value of the genomic data were all above 70%.
Conclusions
Prediction of CHD from genomic data is feasible, and can be applied for risk management of CHD and early selection for genetic improvement to reduce the prevalence of CHD in breeding programs. The prediction can be implemented before maturity, at which age current radiographic screening programs are traditionally applied, and as soon as DNA is available.
doi:10.1016/j.joca.2010.12.011
PMCID: PMC3065507  PMID: 21215318
10.  Identification of Autotoxic Compounds in Fibrous Roots of Rehmannia (Rehmannia glutinosa Libosch.) 
PLoS ONE  2012;7(1):e28806.
Rehmannia is a medicinal plant in China. Autotoxicity has been reported to be one of the major problems hindering the consecutive monoculture of Rehmannia. However, potential autotoxins produced by the fibrous roots are less known. In this study, the autotoxicity of these fibrous roots was investigated. Four groups of autotoxic compounds from the aqueous extracts of the fibrous roots were isolated and characterized. The ethyl acetate extracts of these water-soluble compounds were further analyzed and separated into five fractions. Among them, the most autotoxic fraction (Fr 3) was subjected to GC/MS analysis, resulting in 32 identified compounds. Based on literature, nine compounds were selected for testing their autotoxic effects on radicle growth. Seven out of the nine compounds were phenolic, which significantly reduced radicle growth in a concentration-dependent manner. The other two were aliphatic compounds that showed a moderate inhibition effect at three concentrations. Concentration of these compounds in soil samples was determined by HPLC. Furthermore, the autotoxic compounds were also found in the top soil of the commercially cultivated Rehmannia fields. It appears that a close link exists between the autotoxic effects on the seedlings and the compounds extracted from fibrous roots of Rehmannia.
doi:10.1371/journal.pone.0028806
PMCID: PMC3250401  PMID: 22235251
11.  Therapeutic hypertension system based on a microbreathing pressure sensor system 
Background and methods
A novel therapeutic system for the treatment of hypertension was developed on the basis of a slow-breath training mechanism, using a microbreathing pressure sensor device for the detection of human respiratory signals attached to the abdomen. The system utilizes a single-chip AT89C51 microcomputer as a core processor, programmed by Microsoft Visual C++6.0 to communicate with a PC via a full-speed PDIUSBD12 interface chip. The programming is based on a slow-breath guided algorithm in which the respiratory signal serves as a physiological feedback parameter. Inhalation and exhalation by the subject is guided by music signals.
Results and conclusion
Our study indicates that this microbreathing sensor system may assist in slow-breath training and may help to decrease blood pressure.
doi:10.2147/MDER.S19971
PMCID: PMC3417874  PMID: 22915930
hypertension; microbreathing sensor; single-chip microcomputer; slow-pace breathing
12.  Basolateral Amygdala Inactivation Impairs Learning-Induced Long-Term Potentiation in the Cerebellar Cortex 
PLoS ONE  2011;6(1):e16673.
Learning to fear dangerous situations requires the participation of basolateral amygdala (BLA). In the present study, we provide evidence that BLA is necessary for the synaptic strengthening occurring during memory formation in the cerebellum in rats. In the cerebellar vermis the parallel fibers (PF) to Purkinje cell (PC) synapse is potentiated one day following fear learning. Pretraining BLA inactivation impaired such a learning-induced long-term potentiation (LTP). Similarly, cerebellar LTP is affected when BLA is blocked shortly, but not 6 h, after training. The latter result shows that the effects of BLA inactivation on cerebellar plasticity, when present, are specifically related to memory processes and not due to an interference with sensory or motor functions. These data indicate that fear memory induces cerebellar LTP provided that a heterosynaptic input coming from BLA sets the proper local conditions. Therefore, in the cerebellum, learning-induced plasticity is a heterosynaptic phenomenon that requires inputs from other regions. Studies employing the electrically-induced LTP in order to clarify the cellular mechanisms of memory should therefore take into account the inputs arriving from other brain sites, considering them as integrative units. Based on previous and the present findings, we proposed that BLA enables learning-related plasticity to be formed in the cerebellum in order to respond appropriately to new stimuli or situations.
doi:10.1371/journal.pone.0016673
PMCID: PMC3031621  PMID: 21304962
13.  An intelligent electronic capsule system for automated detection of gastrointestinal bleeding*  
In clinical practice, examination of the hemorrhagic spot (HS) remains difficult. In this paper, we describe a remote controlled capsule (RCC) micro-system with an automated, color-based sensor to identify and localize the HS of the gastrointestinal (GI) tract. In vitro testing of the detecting sensor demonstrated that it was capable of discriminating mimetic intestinal fluid (MIF) with and without the hemoglobin (Hb) when the concentration of Hb in MIF was above 0.05 g/ml. Therefore, this RCC system is able to detect the relatively accurate location of the HS in the GI tract.
doi:10.1631/jzus.B1000047
PMCID: PMC2997402  PMID: 21121072
Smart capsule; Intestinal bleeding; Color sensor; Biomedical micro-electromechanical system (BioMEMS)
14.  Differential Genetic Regulation of Canine Hip Dysplasia and Osteoarthritis 
PLoS ONE  2010;5(10):e13219.
Background
Canine hip dysplasia (HD) is a common polygenic trait characterized by hip malformation that results in osteoarthritis (OA). The condition in dogs is very similar to developmental dysplasia of the human hip which also leads to OA.
Methodology/Principal Findings
A total of 721 dogs, including both an association and linkage population, were genotyped. The association population included 8 pure breeds (Labrador retriever, Greyhounds, German Shepherd, Newfoundland, Golden retriever, Rottweiler, Border Collie and Bernese Mountain Dog). The linkage population included Labrador retrievers, Greyhounds, and their crosses. Of these, 366 dogs were genotyped at ∼22,000 single nucleotide polymorphism (SNP) loci and a targeted screen across 8 chromosomes with ∼3,300 SNPs was performed on 551 dogs (196 dogs were common to both sets). A mixed linear model approach was used to perform an association study on this combined association and linkage population. The study identified 4 susceptibility SNPs associated with HD and 2 SNPs associated with hip OA.
Conclusion/Significance
The identified SNPs included those near known genes (PTPRD, PARD3B, and COL15A1) reported to be associated with, or expressed in, OA in humans. This suggested that the canine model could provide a unique opportunity to identify genes underlying natural HD and hip OA, which are common and debilitating conditions in both dogs and humans.
doi:10.1371/journal.pone.0013219
PMCID: PMC2952589  PMID: 20949002
15.  Glucagon-Like Peptide 1/Glucagon Receptor Dual Agonism Reverses Obesity in Mice 
Diabetes  2009;58(10):2258-2266.
OBJECTIVE
Oxyntomodulin (OXM) is a glucagon-like peptide 1 (GLP-1) receptor (GLP1R)/glucagon receptor (GCGR) dual agonist peptide that reduces body weight in obese subjects through increased energy expenditure and decreased energy intake. The metabolic effects of OXM have been attributed primarily to GLP1R agonism. We examined whether a long acting GLP1R/GCGR dual agonist peptide exerts metabolic effects in diet-induced obese mice that are distinct from those obtained with a GLP1R-selective agonist.
RESEARCH DESIGN AND METHODS
We developed a protease-resistant dual GLP1R/GCGR agonist, DualAG, and a corresponding GLP1R-selective agonist, GLPAG, matched for GLP1R agonist potency and pharmacokinetics. The metabolic effects of these two peptides with respect to weight loss, caloric reduction, glucose control, and lipid lowering, were compared upon chronic dosing in diet-induced obese (DIO) mice. Acute studies in DIO mice revealed metabolic pathways that were modulated independent of weight loss. Studies in Glp1r−/− and Gcgr−/− mice enabled delineation of the contribution of GLP1R versus GCGR activation to the pharmacology of DualAG.
RESULTS
Peptide DualAG exhibits superior weight loss, lipid-lowering activity, and antihyperglycemic efficacy comparable to GLPAG. Improvements in plasma metabolic parameters including insulin, leptin, and adiponectin were more pronounced upon chronic treatment with DualAG than with GLPAG. Dual receptor agonism also increased fatty acid oxidation and reduced hepatic steatosis in DIO mice. The antiobesity effects of DualAG require activation of both GLP1R and GCGR.
CONCLUSIONS
Sustained GLP1R/GCGR dual agonism reverses obesity in DIO mice and is a novel therapeutic approach to the treatment of obesity.
doi:10.2337/db09-0278
PMCID: PMC2750209  PMID: 19602537
16.  The long (and winding) road to gene discovery for canine hip dysplasia 
Hip dysplasia is a common inherited trait of dogs that results in secondary osteoarthritis. In this article the methods used to uncover the mutations contributing to this condition are reviewed, beginning with hip phenotyping. Coarse, genome-wide, microsatellite-based screens of pedigrees of greyhounds and dysplastic Labrador retrievers were used to identify linked quantitative trait loci (QTL). Fine-mapping across two chromosomes (CFA11 and 29) was employed using single nucleotide polymorphism (SNP) genotyping. Power analyses and preferential selection of dogs for ongoing SNP-based genotyping is described with the aim of refining the QTL intervals to 1–2 megabases on these and several additional chromosomes prior to candidate gene screening. The review considers how a mutation or a genetic marker such as a SNP or haplotype of SNPs might be combined with pedigree and phenotype information to create a ‘breeding value’ that could improve the accuracy of predicting a dog’s hip conformation.
doi:10.1016/j.tvjl.2009.02.008
PMCID: PMC2679856  PMID: 19297220
Canine hip dysplasia; Genome wide screen; Microsatellites; Single nucleotide polymorphisms (SNP); Breeding values
17.  Simultaneous detection of different serum pepsinogens and its primary application 
AIM: To develop the simple, rapid and sensitive dual-label time-resolved fluoroimmunoassay for pepsinogens in human serum.
METHODS: Based on two-site sandwich protocol, monoclonal antibodies (McAbs) against pepsinogen I (PG I) and PG II were co-coated in 96 microtitration wells, and tracer McAbs against PG I and PG II were labeled with europium (Eu) and samarium (Sm) chelate, respectively. Diluted serum samples of Eu3+- and Sm3+-McAbs were added into microtitration wells simultaneously. After washing, fluorescence of bound Sm3+ and Eu3+ tracers was detected.
RESULTS: The detection limit was 0.2 μg/L for PG I and 0.05 μg/L for PG II. The assay range was 5.0-320.0 μg/L for PG I and 1.0-55.0 μg/L for PG II. The average recovery rate was 102.7% for PG I and 98.8% for PG II. Sera from healthy controls and patients with gastric disease were analyzed. The PG detected by dual-label assay was in good agreement with that detected by single-label assay or by enzyme-linked immunosorbent assay.
CONCLUSION: Dual-label assay can provide high-throughput serological screening for gastric diseases.
doi:10.3748/wjg.v16.i24.3072
PMCID: PMC2890949  PMID: 20572312
Serum pepsinogen; Simultaneous detection; Time-resolved fluoroimmunoassay
18.  Retrospective Analysis for Genetic Improvement of Hip Joints of Cohort Labrador Retrievers in the United States: 1970–2007 
PLoS ONE  2010;5(2):e9410.
Background
Canine Hip Dysplasia (CHD) is a common inherited disease that affects dog wellbeing and causes a heavy financial and emotional burden to dog owners and breeders due to secondary hip osteoarthritis. The Orthopedic Foundation for Animals (OFA) initiated a program in the 1960's to radiograph hip and elbow joints and release the OFA scores to the public for breeding dogs against CHD. Over last four decades, more than one million radiographic scores have been released.
Methodology/Principal Findings
The pedigrees in the OFA database consisted of 258,851 Labrador retrievers, the major breed scored by the OFA (25% of total records). Of these, 154,352 dogs had an OFA hip score reported between 1970 and 2007. The rest of the dogs (104,499) were the ancestors of the 154,352 dogs to link the pedigree relationships. The OFA hip score is based on a 7-point scale with the best ranked as 1 (excellent) and the worst hip dysplasia as 7. A mixed linear model was used to estimate the effects of age, sex, and test year period and to predict the breeding value for each dog. Additive genetic and residual variances were estimated using the average information restricted maximum likelihood procedure. The analysis also provided an inbreeding coefficient for each dog. The hip scores averaged 1.93 (±SD = 0.59) and the heritability was 0.21. A steady genetic improvement has accrued over the four decades. The breeding values decreased (improved) linearly. By the end of 2005, the total genetic improvement was 0.1 units, which is equivalent to 17% of the total phenotypic standard deviation.
Conclusion/Significance
A steady genetic improvement has been achieved through the selection based on the raw phenotype released by the OFA. As the heritability of the hip score was on the low end (0.21) of reported ranges, we propose that selection based on breeding values will result in more rapid genetic improvement than breeding based on phenotypic selection alone.
doi:10.1371/journal.pone.0009410
PMCID: PMC2827553  PMID: 20195372
19.  A Single IGF1 Allele Is a Major Determinant of Small Size in Dogs 
Science (New York, N.Y.)  2007;316(5821):112-115.
The domestic dog exhibits greater diversity in body size than any other terrestrial vertebrate. We used a strategy that exploits the breed structure of dogs to investigate the genetic basis of size. First, through a genome-wide scan, we identified a major quantitative trait locus (QTL) on chromosome 15 influencing size variation within a single breed. Second, we examined genetic variation in the 15-megabase interval surrounding the QTL in small and giant breeds and found marked evidence for a selective sweep spanning a single gene (IGF1), encoding insulin-like growth factor 1. A single IGF1 single-nucleotide polymorphism haplotype is common to all small breeds and nearly absent from giant breeds, suggesting that the same causal sequence variant is a major contributor to body size in all small dogs.
doi:10.1126/science.1137045
PMCID: PMC2789551  PMID: 17412960
20.  Endoscopic diagnosis of gastrointestinal graft-versus-host disease 
AIM: To evaluate the diagnostic value of endoscopy in patients with gastrointestinal graft-versus-host disease (GI GVHD).
METHODS: We identified 8 patients with GI GVHD following allogeneic hematopoietic stem cell trans-plantation (HSCT). GVHD was defined histologically as the presence of gland apoptosis, not explained by other inflammatory or infectious etiologies.
RESULTS: The symptoms of GI GVHD included anorexia, nausea, vomiting, watery diarrhea, abdominal pain, GI bleeding, etc. Upper endoscopic appearance varied from subtle mucosal edema, hyperemia, erythema to obvious erosion. Colonoscopic examination showed diffuse edema, hyperemia, patchy erosion, scattered ulcer, sloughing and active bleeding. Histological changes in GI GVHD included apoptosis of crypt epithelial cells, dropout of crypts, and lymphocytic infiltration in epithelium and lamina propria. The involvement of stomach and rectocolon varied from diffuse to focal.
CONCLUSION: Endoscopy may play a significant role in early diagnosis of GI GVHD patients following allogeneic HSCT, and histologic examination of gastrointestinal biopsies is needed to confirm the final diagnosis.
doi:10.3748/wjg.14.2262
PMCID: PMC2703857  PMID: 18407606
Gastrointestinal graft-versus-host disease; Endoscopy; Diagnosis; Allogeneic hematopoietic stem cell transplantation
21.  Investigation of iodine concentration in salt, water and soil along the coast of Zhejiang, China*  
Objective: We aim to describe the environment iodine concentration in salt, water and soil along Zhejiang Province coast in the China foreland. It will be helpful for us to judge whether this area is insufficient in iodine and universal iodized salt is necessary or not. Methods: We collected iodized salt samples, drinking water samples (tap water in the towns, and well water or spring water in the villages), water samples from different sources (ditches, lakes, rivers) and soil samples through random sampling in June, 2005. Salt, water and soil iodine was detected by arsenic-cerium redox method. Statistical analysis was expressed as mean±SEM by Windows SPSS 13.0. Results: (1) The iodine concentration in salt was 27.9±4.33 mg/kg (n=108). (2) Seventy-five water samples were collected. The water iodine value was 0.6~84.8 μg/L (mean of 11.66 μg/L). The watershed along the Qiantang River has significantly higher iodine content than the water in Lin’an in mountain area (P<0.01). The iodine content and mean iodine content of tap water, well or spring water and natural water sources were 4.30±2.43 μg/L (n=34), 23.59±27.74 μg/L (n=19) and 12.72±10.72 μg/L (n=22) respectively. This indicated that among environmental water sources, the ditch iodine content was the highest with river water iodine being the lowest (P<0.01). (3) Soil iodine value was 0.11~2.93 mg/kg (mean of 1.32 mg/kg). Though there was no statistical difference of soil iodine in different districts (P=0.131), soil iodine content correlated positively with water iodine content. Conclusion: Iodine concentration in salt accords with national policy of adding iodine in salt. Foreland has more iodine in water than mountain area. The data reflected that water and soil iodine in foreland area was not high, which suggests universal iodized salt should be necessary. Environment iodine has relatively close association with pollution.
doi:10.1631/jzus.2005.B1200
PMCID: PMC1390644  PMID: 16358379
Iodine; Salt; Water; Soil; Coast
22.  Microcalorimetric study on host-guest complexation of naphtho-15-crown-5 with four ions of alkaline earth metal*  
Thermodynamic parameters of complexation of naphto-15-crown-5 with four alkaline earth ions in aqueous media was determined using titration microcalorimetry at 298.15 K. The stability of the complexes, thermal effect and entropy effect of the complexation is discussed on the basis of the guest ions structure and the solvent effect. The stability constants tendency to vary with ion radius was interpreted. Complex of naphtha-15-crown-5 with calcium ion is very stable due to the synergism of static electric interaction and size selectivity between the host and the guest.
doi:10.1631/jzus.2005.B0069
PMCID: PMC1390764  PMID: 15593396
Microcalorimetry; Host-guest complexation; Naphtho-15-crown-5; Alkaline earth metal; Molecular recognition
23.  Evaluation of Cytotoxic T-Lymphocyte Responses in Human and Nonhuman Primate Subjects Infected with Human Immunodeficiency Virus Type 1 or Simian/Human Immunodeficiency Virus 
Journal of Virology  2001;75(1):73-82.
Cytotoxic T-lymphocyte (CTL) responses have been implicated as playing an important role in control of human immunodeficiency virus (HIV) infection. However, it is technically difficult to demonstrate CTL responses consistently in nonhuman primate and human subjects using traditional cytotoxicity assay methods. In this study, we systematically evaluated culture conditions that may affect the proliferation and expansion of CTL effector cells and presented a sensitive method for detection of cytotoxicity responses with bulk CTL cultures. We confirmed the sensitivity and specificity of this method by demonstration of vigorous CTL responses in a simian-HIV (SHIV)-infected rhesus macaque. The expansion of epitope-specific CTL effector cells was also measured quantitatively by CTL epitope-major histocompatibility complex tetramer complex staining. In addition, two new T-cell determinants in the SIV gag region are identified. Last, we showed the utility of this method for studying CTL responses in chimpanzee and human subjects.
doi:10.1128/JVI.75.1.73-82.2001
PMCID: PMC113899  PMID: 11119575

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