• Title/Summary/Keyword: 2-cell block

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Channeling of Intermediates Derived from Medium-Chain Fatty Acids and De novo-SYnthesized Fatty Acids to Polyhydroxyalkanoic Acid by 2-Bromooctanoic Acid in Pseudomonas fluorescens BM07

  • LEE, HO-JOO;RHO, JONG-KOOK;KAMBIZ AKBARI NOGHABI,;LEE, SEUNG-EUN;CHOI, MUN-HWAN;YOON, SUNG-CHUL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1256-1266
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    • 2004
  • 2-Bromooctanoic acid (2-BrOA) is known to block the formation of polyhydroxyalkanoic acid (PHA) in Pseudomonasfluorescens BM07 without any influence on the cell growth when grown on fructose, but it inhibits the cell growth when grown on octanoate (OA) (Lee et al., Appl. Environ. Microbiol. 67: 4963- 4974, 2001). We investigated the role of 2-BrOA in the PHA synthesis of the bacterium grown with mixtures of fructose and fatty acids. OA, 11­phenoxyundecanoic acid (1 1-POU), and 5-phenylvaleric acid (5-PV) were selected as model substrates. When supplemented with 50 mM fructose, all these carboxylic acids suppressed the formation of PHA from fructose, however, the ~-oxidation coenzyme A monomers derived from the carboxylic acids were efficiently polymerized, but the conversion yield [(mol of carboxylate substrate converted into PHA)/(mol of carboxylate substrate in the feed)] was low (e.g., maximally $\~53\%$ for 5 mM 11-POU). Addition of 2-BrOA (up to 5 mM) to the mixed carbon sources raised the conversion yield sensitively and effectively only at low levels of the acid substrates (e.g., 2 mM 1 1-POU or 5 mM OA): For instance, $100\%$ of 2 mM ll-POU were converted into PHA in the presence of 5 mM 2-BrOA, whereas only $\~10\%$ of the 1 1-POU were converted in the absence of 2-BrOA. However, at highly saturated suppressing levels (e.g., 5 mM ll-POU), 2-BrOA inhibitor showed no significant additional effect on the conversion ($60- 70\%$ conversion irrespective of 2-BrOA level). The existence of competitive and compensative relationship between 2­BrOA and all the carboxylic acid substrates used may indicate 'Present address: Section on Brain Physiology and Metabolism, Bldg. 10, Rm. 6N202, National Institute on Agmg, National Institute of Health, Bethesda, MD 20892, U.S.A. that all the acid substrate-derived inhibiting species bind to the same site as the 2-BrOA inhibiting species does. We, therefore, suggest that 2-BrOA can be used for efficiently increasing the yield of conversion of expensive substituted fatty acids into PHA and then substituted 3-hydroxyacids by hydrolyzing it.

Electrical Stimulation Parameters in Normal and Degenerate Rabbit Retina (정상 망막과 변성 망막을 위한 전기자극 파라미터)

  • Jin, Gye-Hwan;Goo, Yong-Sook
    • Progress in Medical Physics
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    • v.19 no.1
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    • pp.73-79
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    • 2008
  • Retinal prosthesis is regarded as the most feasible method for the blind caused by retinal diseases such as retinitis pigmentosa (RP) or age related macular degeneration (AMD). Recently Korean consortium launched for developing retinal prosthesis. One of the prerequisites for the success of retinal prosthesis is the optimization of the electrical stimuli applied through the prosthesis. Since electrical characteristics of degenerate retina are expected to differ from those of normal retina, we performed voltage stimulation experiment both in normal and degenerate retina to provide a guideline for the optimization of electrical stimulation for the upcoming prosthesis. After isolation of retina, retinal patch was attached with the ganglion cell side facing the surface of microelectrode arrays (MEA). $8{\times}8$ grid layout MEA (electrode diameter: $30{\mu}m$, electrode spacing: $200{\mu}m$, and impedance: $50k{\Omega}$ at 1 kHz) was used to record in-vitro retinal ganglion cell activity. Mono-polar electrical stimulation was applied through one of the 60 MEA channel, and the remaining channels were used for recording. The electrical stimulus was a constant voltage, charge-balanced biphasic, anodic-first square wave pulse without interphase delay, and 50 trains of pulse was applied with a period of 2 sec. Different electrical stimuli were applied. First, pulse amplitude was varied (voltage: $0.5{\sim}3.0V$). Second, pulse duration was varied $(100{\sim}1,200{\mu}s)$. Evoked responses were analyzed by PSTH from averaged data with 50 trials. Charge density was calculated with Ohm's and Coulomb's law. In normal retina, by varying the pulse amplitude from 0.5 to 3V with fixed duration of $500{\mu}s$, the threshold level for reliable ganglion cell response was found at 1.5V. The calculated threshold of charge density was $2.123mC/cm^2$. By varying the pulse duration from 100 to $1,200{\mu}s$ with fixed amplitude of 2V, the threshold level was found at $300{\mu}s$. The calculated threhold of charge density was $1.698mC/cm^2$. Even after the block of ON-pathway with L-(1)-2-amino-4-phosphonobutyric acid (APB), electrical stimulus evoked ganglion cell activities. In this APB-induced degenerate retina, by varying the pulse duration from 100 to $1200{\mu}s$ with fixed voltage of 2 V, the threshold level was found at $300{\mu}s$, which is the same with normal retina. More experiment with APB-induced degenerate retina is needed to make a clear comparison of threshold of charge density between normal and degenerate retina.

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Effect of Dopamine on the $Ca^{2+}\;-dependent\;K^+\;currents$ in Isolated Single Gastric Myocytes of the Guinea-pig

  • Rhee, Poong-Lyul;Lee, Sang-Jin;Kim, Sung-Joon;So, In-Suk;Hwang, Sang-Ik;Kim, Ki-Whan
    • The Korean Journal of Physiology
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    • v.27 no.2
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    • pp.139-150
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    • 1993
  • We have reported that dopamine potentiates spontaneous contractions dose-dependently in guinea-pig antral circular muscle strips (Hwang et al, 1991). To clarify the underlying excitatory mechanism of dopamine on the gastric smooth muscle, the effects of dopamine on voltage-dependent $Ca^{2+}\;currents\;and\;Ca^{2+}\;-dependent\;K^+\;currents$ were observed in enzymatically dispersed guinea-pig gastric myocytes using the whole-cell voltage-clamp technique. Experiments were also done using isometric tension recording and conventional intracellular microelectrode techniques. 1) The effect of dopamine on the spontaneous contraction of antral circular muscle strips of the guinea-pig was excitatory in a dose-dependent manner, and was blocked by phentolamine, an ${\alpha}-adrenoceptor$ blocker. 2) The slow waves were not changed by dopamine. 3) The voltage-operated inward $Ca^{2+}$ current was not influenced by dopamine. 4) The $Ca^{2+}\;-dependent\;K^+$ outward current, which might reflect the changes of intracellular calcium concentration, was enhanced by dopamine. This effect was abolished by phentolamine. 5) The enhancing effect of dopamine on the $Ca^{2+}\;-dependent\;K^+$ current disappeared with heparin which is known to block the action of $InsP_3$. From these results, it is suggested that dopamine acts via $InsP_3-mediated\;Ca^{2+}$ mobilization from intracellular stores and such action potentiates the spontaneous contraction of guinea-pig gastric smooth muscle.

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Effects of Histamine on Cultured Interstitial Cells of Cajal in Murine Small Intestine

  • Kim, Byung Joo;Kwon, Young Kyu;Kim, Euiyong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.2
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    • pp.149-156
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    • 2013
  • Interstitial cells of Cajal (ICCs) are the pacemaker cells in the gastrointestinal tract, and histamine is known to regulate neuronal activity, control vascular tone, alter endothelial permeability, and modulate gastric acid secretion. However, the action mechanisms of histamine in mouse small intestinal ICCs have not been previously investigated, and thus, in the present study, we investigated the effects of histamine on mouse small intestinal ICCs, and sought to identify the receptors involved. Enzymatic digestions were used to dissociate ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record potentials (in current clamp mode) from cultured ICCs. Histamine was found to depolarize resting membrane potentials concentration dependently, and whereas 2-PEA (a selective H1 receptor agonist) induced membrane depolarizations, Dimaprit (a selective H2-agonist), R-alpha-methylhistamine (R-alpha-MeHa; a selective H3-agonist), and 4-methylhistamine (4-MH; a selective H4-agonist) did not. Pretreatment with $Ca^{2+}$-free solution or thapsigargin (a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum) abolished the generation of pacemaker potentials and suppressed histamine-induced membrane depolarization. Furthermore, treatments with U-73122 (a phospholipase C inhibitor) or 5-fluoro-2-indolyl des-chlorohalopemide (FIPI; a phospholipase D inhibitor) blocked histamine-induced membrane depolarizations in ICCs. On the other hand, KT5720 (a protein kinase A inhibitor) did not block histamine-induced membrane depolarization. These results suggest that histamine modulates pacemaker potentials through H1 receptor-mediated pathways via external $Ca^{2+}$ influx and $Ca^{2+}$ release from internal stores in a PLC and PLD dependent manner.

The Correlation between Caudal Epidurogram and Low Back Pain

  • Jo, Dae-Hyun;Jang, Sul
    • The Korean Journal of Pain
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    • v.25 no.1
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    • pp.22-27
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    • 2012
  • Background: The common causes of lower back pain with or without leg pain includes disk disease and spinal stenosis. A definitive diagnosis is usually made by means of magnetic resonance imaging (MRI), but treatment is often difficult because the MRI findings are not consistent with the symptoms of the patient in many cases. The objective of this study was to observe the correlation between the patterns of epidurography performed in patients having lower back pain with or without leg pain and the position or severity of the pain as subjectively described by the patients. Methods: The subjects of this study were 69 outpatients with lower back pain with or without leg pain who visited our clinic and complained of predominant pain on one side. We performed caudal epidural block using an image intensifier. A mixture of the therapeutic drug and the contrast agent (10 ml) was injected to observe the contrast flow pattern. The patients who complained of predominant pain on one side were divided into the left side group and the right side group. A judgment of inconsistency was made if the contrast agent flowed to the side of the pain, while a judgment of consistency was made if the contrast agent flowed to the opposite side of the pain. The degree of the drug distribution was evaluated by counting the number of cells to which the contrast agent's flowed for evaluating the correlation between the contrasted cell and the severity of pain (one group ${\leq}$ VAS 7, the other group ${\geq}$ VAS 8) the degree of the contrast agent's contrast was evaluated by dividing and counting an image into 15 cells (the left, right, and middle sections at each level of L4, L5, S1, S2, and S3). Results: Thirty out of the 69 patients who had laterality in pain, that is, those who complained of predominant pain on one side, showed that the laterality of the pain and the contrast agent flow was consistent, while 39 patients showed that the laterality was inconsistent (P: 0.137). The evaluation of the correlation between the pain and the contrast agent flow showed that the mean number of contrasted cells was $9.0{\pm}2.2$ for the 46 patients in the group with a VAS of 7 or lower and $6.5{\pm}2.0$ for the 23 patients in the group with a VAS of 8 or higher, indicating that the former group showed a significantly greater number of contrasted cells (P < 0.001). Conclusions: This study, conducted with patients having lower back pain with or without leg pain, showed that the contrast flow pattern of caudal epidurography had a significant correlation with the severity of the pain but not with the laterality of the pain.

The Significance of SDF-1α-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells

  • Bae, Yoon-Kyung;Kim, Gee-Hye;Lee, Jae Cheoun;Seo, Byoung-Moo;Joo, Kyeung-Min;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.40 no.6
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    • pp.386-392
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    • 2017
  • Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

Inhibitory Effects of Deer Antler Herbal-Acupuncture Solution on Cathepsin S and L Activity in RA Model Mice

  • Ahn, Hyung-Jun;Yoon, Jong-Hwa;Kim, Kyung-Ho;Lee, Seung-Deok;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.71-81
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    • 2005
  • Objective : Dear antler (Cervus korean TEMMINCK var. mantchuricus Swinhoe) used for traditional immunosuppressive and immuno-activating action. The effect of deer antler herbal-acupuncture(DAH) solution, prepared by water extract method, on cathepsin activities in bone tissues (cartilage and synovial) cells from mouse rheumatoid arthritis (RA) model was studied. The cysteine endoprotease cathepsin mediates degradation of the MHC class II invariant chain (Ii) in human and mouse antigen-presenting cells. The studies described here examine the functional significance of cathepsin inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for RA. Methods : An animal model for RA in BALB/c mice thymectomized 3 days after birth (3d-Tx) was constructed All 3d-Tx BALB/c mice developed autoimmune lesions in the bone tissue cells, starting at 3 weeks of age, and the disease mediated by CD4+ T cells was chronic and progressive. Significant inhibitory effects of DAH solution on cathepsin S and L were observed in each organ in a dose-dependent manner. Moreover, we confirmed that cathepsin S and L activity in each organ were clearly inhibited by DAB solution. When we examined the inhibitory effects of DAH solution against autoantigen-specific T cell responses in vitro, in regional lymph node cells, but not in spleens, from model mice, a significant inhibitory effect of DAB solution was observed in a dose-dependent manner. DAH solution do not block T cell proliferation to Con A, indicated that the dose of DAB solution 10 to $20\;{\mu}g/m{\ell}$ was sufficient to inactivate the autoantigen-specific T cell responses in vitro. In vivo therapeutic effects of DAB solution were examined in a murine model for RA, autoantigen-specific (C-II-specific) T cell response were significantly inhibited in LNCs from DAH solution-treated mice. Results : Iinhibition of cathepsin S and L in vivo alters autoantigen presentation and development of organ-specific autoimmunity in RA model. Conclusion : These data identify selective inhibition of cysteine protease cathepsin S and L as a potential therapeutic strategy for autoimmune disease process such RA. Thus, DAH solution will served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

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Spectrum Reuse Schemes with Power Control for Device-to-Device Communication in LTE-Advanced Cellular Network

  • Chhorn, Sok;Yoon, Seok-Ho;Seo, Si-O;Kim, Seung-Yeon;Cho, Choong-Ho
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.9 no.12
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    • pp.4819-4834
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    • 2015
  • The spectral efficiency of cellular networks can be improved when proximate users engage in device-to-device (D2D) communications to communicate directly without going through a base station. However, D2D communications that are not properly designed may generate interference with existing cellular networks. In this paper, we study resource allocation and power control to minimize the probability of an outage and maximize the overall network throughput. We investigate three power control-based schemes: the Partial Co-channel based Overlap Resource Power Control (PC.OVER), Fractional Frequency Reuse based Overlap Resource Power Control (FFR.OVER) and Fractional Frequency Reuse based Adaptive Power Control (FFR.APC) and also compare their performance. In PC.OVER, a certain portion of the total bandwidth is dedicated to the D2D. The FFR.OVER and FFR.APC schemes combine the FFR techniques and the power control mechanism. In FFR, the entire frequency band is partitioned into two parts, including a central and edge sub-bands. Macrocell users (mUEs) transmit using uniform power in the inner and outer regions of the cell, and in all three schemes, the D2D receivers (D2DRs) transmit with low power when more than one D2DRs share a resource block (RB) with the macrocells. For PC.OVER and FFR.OVER, the power of the D2DRs is reduced to its minimum, and for the FFR.APC scheme, the transmission power of the D2DRs is iteratively adjusted to satisfy the signal to interference ratio (SIR) threshold. The three schemes exhibit a significant improvement in the overall system capacity as well as in the probability of a user outage when compared to a conventional scheme.

BONE HEALING CAPACITY OF THE COLLAGEN BONE FILLER ($TERUPLUG^{(R)}$) AND RHBMP-2 IN THE RABBIT CRANIUM DEFECT (가토 두개골 결손부에 이식된 Collagen bone filler ($TERUPLUG^{(R)}$) 및 rhBMP-2의 골치유 능력)

  • Kim, Ju-Hoon;Kim, Chul-Hwan;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.2
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    • pp.119-130
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    • 2008
  • Absorbable atelo-collagen sponge $TERUPLUG^{(R)}$, Termo Co. Tokyo, Japan) is inserted in the extraction wound where alveolar bone is exposed. It protects wounds and promotes the formation of granulation. This is made of atelo-collagen, to minimize antigenicity, which is cross-linked by heat treatment for biocompatibility. $TERUPLUG^{(R)}$ consists of between 85 and 95 % of collagen type I and between 5 to 15 % of collagen type III. The raw material for the collagen is derived from bovine skin. It features a sponge block design and is shaped for easy insertion in the extraction wound. This study was designed to find out the bone healing capacity of $TERUPLUG^{(R)}$. We implanted $TERUPLUG^{(R)}$ (experimental group I) and $TERUPLUG^{(R)}$ with rhBMP-2 (experimental group II) in the rabbit cranium defect and then histologically analysed the specimen. The results were as follows. 1. In the 4 weeks, a lot of the newly formed collagen fibers around material of the experimental group I implanted $TERUPLUG^{(R)}$ were observed. But, in the experimental group II implanted $TERUPLUG^{(R)}$ with rhBMP-2, a little of newly formed collagen fibers around material were observed. The cell proliferating activity and apoptosis of the experimental group I, II was positive in and around the implanted material. 2. In the 8 weeks, the amount of newly formed and matured bone in the experimental group II was more observed than the experimental group I and control group. The results of this study indicate that absorbable atelo-collagen sponge ($TERUPLUG^{(R)}$) is relatively favorable bone void filler with biocompatibility and has the better bone healing capacity in case of application with rhBMP-2.

The venom of jellyfish, Chrysaora pacifica, induces neurotoxicity via activating Ca2+-mediated ROS signaling in HT-22 cells

  • Yang, Yoon-Sil;Kang, Young-Joon;Kim, Hye-Ji;Kim, Min-Soo;Jung, Sung-Cherl
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.347-353
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    • 2019
  • Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.