Epidermal Growth Factor Stimulates Cell Proliferation by Activating Voltage-Gated Potassium Channels in Rat Bone Marrow-Derived Mesenchymal Stem Cells

Poster PresentationBackground and objective: We have previously found that voltage-gated delayed rectifier potassium current (IKDR, encoded by Kv1.2 and Kv2.1) participated in regulation of cell cycling progression in rat mesenchymal stem cells (MSCs) from bone marrow. The present study was designed to investigate whether epidermal growth factor (EGF) regulates cell growth is mediated by activating IKDR. Methods: Whole-cell patch voltage-clamp, RT-PCR, Western blots, siRNA, cell proliferation assay were employed in the present study Results: EGF increased cell proliferation in a concentration-dependent manner, and the effect was countered by the broad spectrum protein tyrosine (PTK) inhibitor genistein and the EGFR kinase inhibitor AG556. We found that genistein and AG556 inhibited IKDR in a concentration-dependent manner, The protein tyrosine phosphatase (PTP) inhibitor orthovanadate enhanced IKDR, and counted the inhibitory effect of IKDR by genistein or AG556, suggesting the PTK-mediating modulation of IKDR. Interestingly EGF also increased IKDR, Downregulation of IKDR with siRNA targeting to Kv1.2 or Kv2.1 channels inhibited basal proliferation, and prevented EGF-stimulated proliferation in rat MSCs. Conclusion: These results demonstrate for the first time that EGF stimulates cell proliferation activating IKDR, and silencing Kv1.2 or Kv2.1 channels prevents the augmentation of proliferation by EFG, indicating that Kv1.2 and Kv2.1 channels mediate EGF effect in regulating cell growth in rat MSCs.published_or_final_versio

Non-thermal plasma technology for the development of antimicrobial surfaces: a review

Antimicrobial coatings are in high demand in many fields including the biomaterials and healthcare sectors. Within recent progress in nanoscience and engineering at the nanoscale, preparation of nanocomposite films containing metal nanoparticles ( such as silver nanoparticles, copper nanoparticles, zinc oxide nanoparticles) is becoming an important step in manufacturing biomaterials with high antimicrobial activity. Controlled release of antibiotic agents and eliminating free nanoparticles are of equal importance for engineering antimicrobial nanocomposite materials. Compared to traditional chemical 'wet' methods, plasma deposition and plasma polymerization are promising approaches for the fabrication of nanocomposite films with the advantages of gas phase dry processes, effective use of chemicals and applicability to various substrates. In this article, we present a short overview of state-of-the-art engineering of antimicrobial materials based on the use of non-thermal plasmas at low and atmospheric pressure

EGFR Tyrosine kinase regulates small conductance Ca2+-activated K+ (hSKCa1) channels expressed in HEK 293 cells

SKCa (small-conductance Ca(2+)-activated K(+)) channels are widely distributed in different tissues, including the brain, pancreatic islets and myocardium and play an important role in controlling electrical activity and cellular functions. However, intracellular signal modulation of SKCa channels is not fully understood. The present study was designed to investigate the potential regulation of hSKCa1 (human SKCa1) channels by PTKs (protein tyrosine kinases) in HEK (human embryonic kidney)-293 cells expressing the hSKCa1 (KCNN1) gene using approaches of whole-cell patch voltage-clamp, immunoprecipitation, Western blotting and mutagenesis. We found that the hSKCa1 current was inhibited by the broad-spectrum PTK inhibitor genistein, the selective EGFR (epidermal growth factor receptor) kinase inhibitors T25 (tyrphostin 25) and AG556 (tyrphostin AG 556), but not by the Src-family kinases inhibitor PP2. The inhibitory effect of these PTK inhibitors was significantly antagonized by the PTP (protein tyrosine phosphatase) inhibitor orthovanadate. The tyrosine phosphorylation level of hSKCa1 channels was reduced by genistein, T25 or AG556. The reduced tyrosine phosphorylation was countered by orthovanadate. Interestingly, the Y109F mutant hSKCa1 channel lost the inhibitory response to T25 or AG556, and showed a dramatic reduction in tyrosine phosphorylation levels and a reduced current density. These results demonstrate the novel information that hSKCa1 channels are inhibited by genistein, T25 and AG556 via EGFR tyrosine kinase inhibition, which is related to the phosphorylation of Tyr(109) in the N-terminus. This effect may affect electrical activity and cellular functions in brain, pancreatic islets and myocardium.postprin

Modulation of human cardiac transient outward potassium current by EGFR tyrosine kinase and Src-family kinases

Aims: The human cardiac transient outward K + current Ito (encoded by Kv4.3 or KCND3) plays an important role in phase 1 rapid repolarization of cardiac action potentials in the heart. However, modulation of I to by intracellular signal transduction is not fully understood. The present study was therefore designed to determine whether/how human atrial I to and hKv4.3 channels stably expressed in HEK 293 cells are regulated by protein tyrosine kinases (PTKs). Methods and results: Whole-cell patch voltage-clamp, immunoprecipitation, western blotting, and site-directed mutagenesis approaches were employed in the present study. We found that human atrial I to was inhibited by the broad-spectrum PTK inhibitor genistein, the selective epidermal growth factor receptor (EGFR) kinase inhibitor AG556, and the Src-family kinases inhibitor PP2. The inhibitory effect was countered by the protein tyrosine phosphatase inhibitor orthovanadate. In HEK 293 cells stably expressing human KCND3, genistein, AG556, and PP2 significantly reduced the hKv4.3 current, and the reduction was antagonized by orthovanadate. Interestingly, orthovanadate also reversed the reduced tyrosine phosphorylation level of hKv4.3 channels by genistein, AG556, or PP2. Mutagenesis revealed that the hKv4.3 mutant Y136F lost the inhibitory response to AG556, while Y108F lost response to PP2. The double-mutant Y108FY136F hKv4.3 channels showed no response to either AG556 or PP2. Conclusion: Our results demonstrate that human atrial Ito and cloned hKv4.3 channels are modulated by EGFR kinase via phosphorylation of the Y136 residue and by Src-family kinases via phosphorylation of the Y108 residue; tyrosine phosphorylation of the channel may be involved in regulating cardiac electrophysiology. © The Author 2011.postprin

Genes Underlying Positive Influence Of Prenatal Environmental Enrichment And Negative Influence Of Prenatal Earthquake Simulation And Corrective Influence Of Chinese Herbalmedicine On Rat Offspring: Irf7 And Ninj2

Background: Prenatal environmental enrichment (EE) has been proven to positively affect but prenatal stress negatively influence the physiological and psychological processes in animals, whose trans-generational genetic mechanism remains unclearly defined. We aimed to investigate and find out key genes underlying the positive-negative effects derived from prenatal interventions.Materials and Methods: Pregnant rats were randomized into EE group (EEG), earthquake simulation group (ESG), herbal group (HG) received herbal supplements in feed after earthquake simulation, and control group (CG).Results: Light Box Defecation Test (LBDT) showed EEG offspring presented less fecal pellets than CG offspring, ESG’s more than CG’s, and HG’s less than ESG (p’s<0.05). Open-field Test (OFT) score of EEG was higher than CG offspring, of ESG’s was lower than CG’s, and HG’s higher than ESG’s. Irf7 and Ninj were screened, which were up-regulated in EEG, down-regulated in ESG (FC<0.5), and were neutralized in HG. Prenatal EE could positively promote the nervous system development, prenatal earthquake simulation could retard the nervous system development and Chinese herbal remedy (JKSQW) which could correct the retardation.Conclusion: The negative-positive prenatal effect could contribute to altered gene expression of Irf7 and Ninj2 which also could play a key role in the improving function of JKSQWfor the kidneys.Keywords: Prenatal stress; Earthquake simulation; Light Box Defecation Test; Open-field Test; Irf7; Ninj

Passively Q-switched Yb : YAG laser with Cr4+: YAG as the saturable absorber

By using a continuous-wave Ti:sapphire laser as a pumping source, we demonstrated a passively Q-switched Yb:YAG laser at room temperature with Cr4+:YAG as the saturable absorber. We achieved an average output power of as much as 55 mW at 1.03 mum with a pulse width (FWHM) as short as 350 ns. The initial transmission of the Cr4+:YAG has an effect on the pulse duration (FWHM) and the repetition rate of the Yb:YAG passively Q-switched laser. The Yb:YAG crystal can be a most promising passively Q-switched laser crystal for compact, efficient, solid-state lasers. (C) 2001 Optical Society of America

Search for K_S K_L in psi'' decays

K_S K_L from psi'' decays is searched for using the psi'' data collected by BESII at BEPC, the upper limit of the branching fraction is determined to be B(psi''--> K_S K_L) < 2.1\times 10^{-4} at 90% C. L. The measurement is compared with the prediction of the S- and D-wave mixing model of the charmonia, based on the measurements of the branching fractions of J/psi-->K_S K_L and psi'-->K_S K_L.Comment: 5 pages, 1 figur

Reduced expression of N-Myc downstream-regulated gene 2 in human thyroid cancer

<p>Abstract</p> <p>Background</p> <p><it>NDRG</it>2 (N-Myc downstream-regulated gene 2) was initially cloned in our laboratory. Previous results have shown that <it>NDRG</it>2 expressed differentially in normal and cancer tissues. Specifically, <it>NDRG</it>2 mRNA was down-regulated or undetectable in several human cancers, and over-expression of <it>NDRG</it>2 inhibited the proliferation of cancer cells. <it>NDRG</it>2 also exerts important functions in cell differentiation and tumor suppression. However, it remains unclear whether <it>NDRG</it>2 participates in carcinogenesis of the thyroid.</p> <p>Methods</p> <p>In this study, we investigated the expression profile of human <it>NDRG</it>2 in thyroid adenomas and carcinomas, by examining tissues from individuals with thyroid adenomas (n = 40) and carcinomas (n = 35), along with corresponding normal tissues. Immunohistochemistry, quantitative RT-PCR and western blot methods were utilized to determine both the protein and mRNA expression status of Ndrg2 and c-Myc.</p> <p>Results</p> <p>The immunostaining analysis revealed a decrease of Ndrg2 expression in thyroid carcinomas. When comparing adenomas or carcinomas with adjacent normal tissue from the same individual, the mRNA expression level of <it>NDRG</it>2 was significantly decreased in thyroid carcinoma tissues, while there was little difference in adenoma tissues. This differential expression was confirmed at the protein level by western blotting. However, there were no significant correlations of <it>NDRG</it>2 expression with gender, age, different histotypes of thyroid cancers or distant metastases.</p> <p>Conclusion</p> <p>Our data indicates that <it>NDRG</it>2 may participate in thyroid carcinogenesis. This finding provides novel insight into the important role of <it>NDRG2 </it>in the development of thyroid carcinomas. Future studies are needed to address whether the down-regulation of <it>NDRG</it>2 is a cause or a consequence of the progression from a normal thyroid to a carcinoma.</p

Observation of a ppb mass threshoud enhancement in \psi^\prime\to\pi^+\pi^-J/\psi(J/\psi\to\gamma p\bar{p}) decay

The decay channel $\psi^\prime\to\pi^+\pi^-J/\psi(J/\psi\to\gamma p\bar{p})$ is studied using a sample of $1.06\times 10^8$ $\psi^\prime$ events collected by the BESIII experiment at BEPCII. A strong enhancement at threshold is observed in the $p\bar{p}$ invariant mass spectrum. The enhancement can be fit with an $S$-wave Breit-Wigner resonance function with a resulting peak mass of $M=1861^{+6}_{-13} {\rm (stat)}^{+7}_{-26} {\rm (syst)} {\rm MeV/}c^2$ and a narrow width that is $\Gamma<38 {\rm MeV/}c^2$ at the 90% confidence level. These results are consistent with published BESII results. These mass and width values do not match with those of any known meson resonance.Comment: 5 pages, 3 figures, submitted to Chinese Physics