1.Nidogen Plays a Role in the Regenerative Axon Growth of Adult Sensory Neurons Through Schwann Cells.
Hyun Kyoung LEE ; In Ae SEO ; Duk Joon SUH ; Hwan Tae PARK
Journal of Korean Medical Science 2009;24(4):654-659
We previously reported that nidogen is an extracellular matrix protein regulating Schwann cell proliferation and migration. Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells. Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent. Continuous infusion of recombinant ectodomain of tumor endothelial marker 7, which specifically blocks nidogen function in Schwann cells, suppressed regenerative neurite growth in a sciatic nerve axotomy model. Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.
Animals
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Axotomy
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Cell Movement
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Cell Proliferation
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Male
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Membrane Glycoproteins/*physiology
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Membrane Proteins/pharmacology
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*Nerve Regeneration
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Nerve Tissue Proteins/pharmacology
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Neurites/drug effects/*physiology/ultrastructure
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Rats
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Rats, Sprague-Dawley
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Recombinant Proteins/pharmacology
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Schwann Cells/cytology/*physiology
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Sensory Receptor Cells/*physiology
2.Investigation of optimum concentrations of betaine for improving the resolution of sequencing G-C rich DNA with trinucleotide repeats.
Chinese Journal of Medical Genetics 2014;31(2):163-169
OBJECTIVETo develop an optimal sequencing system which can improve the resolution of sequencing G-C rich DNA with abundant trinucleotide repeats by applying concentration gradients of betaine to the Sanger sequencing system.
METHODSConcentration gradients of betaine were introduced into the sequencing system by taking the 5' terminal of Nogo-B cDNA (Am-Nogo-B) (G-C%=72%, without trinucleotide repeats) and 5' terminal of Huntingtin cDNA (Am-HTT) (G-C%=74%, with abundant CAG and CCG repeats) the results of sequencing were compared.
RESULTSThe optimum concentration of betaine for sequencing Am-Nogo-B has differed from that for Am-HTT. Result of sequencing Am-Nogo-B has achieved the best quality when the concentration of betaine was at 0.8-1.2 mol/L, whereas the result of sequencing Am-HTT obtained the best quality when the concentration of betaine was at 1.6 -2.4 mol/L. The results were reproducible.
CONCLUSIONG-C rich DNA with similar G-C% required different concentrations of betaine in the sequencing system due to base pair compositions. The sequencing system developed for improving the resolution of sequencing of G-C rich DNA with abundant trinucleotide repeats can be used as a reference for similar studies.
Base Sequence ; Betaine ; pharmacology ; Huntingtin Protein ; Molecular Sequence Data ; Nerve Tissue Proteins ; genetics ; Sequence Analysis, DNA ; methods ; Trinucleotide Repeats
3.Effect of 2,5-hexanedione on light-molecular-weight neurofilaments (NF-L) degradation of rat nerve tissues.
Chao-shuang ZOU ; Ke-qin XIE ; Rui-rui KOU ; Yuan GAO ; Fu-yong SONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(6):418-422
OBJECTIVETo investigate the effect of 2,5-hexanedione (HD) on degradation of low-molecular-weight neurofilaments (NF-L) in nervous tissue of rats, and to explore the molecular mechanism of n-hexane neuropathy.
METHODSFifty male Wistar rats were randomly divided into one-week poisoning group (n = 10), two-week poisoning group (n = 10), three-week poisoning group (n = 10), four-week poisoning group (n = 10), and control group (n = 10). In the four poisoning groups, a rat model of n-hexane neuropathy was established by intraperitoneal injection of HD (400 mg/kg/d). The change in the sciatic nerve ultrastructure of each rat was observed under an electron microscope. The progression of HD-induced peripheral neuropathy was evaluated using a gait scoring system. The degradation rates of NF-L in the sciatic nerve and spinal cord of each rat were measured by Western Blotting.
RESULTSThe rats showed decrease in muscle strength and abnormal gait after two weeks of HD poisoning and mild or moderate paralysis after four weeks of HD poisoning. The sciatic nerve showed degenerative change, according to electron microscope observation. Compared with the control group, the two-week poisoning group, three-week poisoning group, and four-week poisoning group had the NF-L degradation rates decreased by 25.8%, 70.4%, and 69.7%, respectively, in the supernatant fraction of sciatic nerve, and by 14.7%, 64.6%, and 67.3%, respectively, in the sediment fraction of sciatic nerve, all showing a significant difference (P < 0.01). Compared with the control group, the one-week poisoning group had the NF-L degradation rate decreased by 33.87% in the supernatant fraction of spinal cord, the four-week poisoning group had the NF-L degradation rate increased by 16.2% in the supernatant fraction of spinal cord, and the one-week poisoning group and two-week poisoning group had the NF-L degradation rates decreased by 46.3% and 13.0% in the sediment fraction of spinal cord, all showing a significant difference (P < 0.01).
CONCLUSIONHD poisoning significantly inhibits NF-L degradation in the sciatic nerve, which may be associated with NF degeneration and accumulation in the axons of patients with n-hexane neuropathy.
Animals ; Hexanes ; poisoning ; Hexanones ; pharmacology ; Male ; Nerve Tissue ; drug effects ; metabolism ; physiopathology ; Neurofilament Proteins ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; metabolism ; physiopathology
4.Affection of metallothionein-3 to dUTPase's accommodating cellular toxicity of dUTP.
Qiao-Lin CHEN ; Qiao-Hua KANG ; Hong-Wei REN ; Zong-Yuan WANG ; Bing-Gen RU
Chinese Journal of Biotechnology 2004;20(3):389-393
Metallothionein-3 (MT-3), renamed as growth inhibitory factor (GIF), is a brain specific member of the metallothionein family. Human dUTPase is a recently found protein in brain that can interact with hMT-3. They have the growth inhibitory activity on neuron cell by interaction. To study the affection of hMT-3 to dUTPase's eliminating the cellular toxicity caused by dUTP, the pSVHA-dUTPase and pFLag-hMT-3 genes have been transfected into HEK293 cells. In addition, the dUTPase and hMT-3 proteins were expressed in BL21 to study the role of hMT-3 on the hydrolyzation of dUTP by dUTPase. The results demonstrate that the cells co-transfected with dUTPase and hMT-3 genes have more strong resistibility to dUTP than the cells transfected only with dUTPase gene. And that the hMT-3 protein can accelerate the hydrolyzation of dUTP by dUTPase. All these indicate that hMT-3 can cooperate with dUTPase to protect better the 293 cells from dUTP. This research offered the theoretic elements for the application of hMT-3 and dUTPase in chemic cure.
Cell Line
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Deoxyuracil Nucleotides
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antagonists & inhibitors
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chemistry
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Nerve Tissue Proteins
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chemistry
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genetics
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pharmacology
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Neurons
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cytology
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drug effects
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Protein Interaction Domains and Motifs
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Pyrophosphatases
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chemistry
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genetics
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pharmacology
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Recombinant Proteins
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biosynthesis
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genetics
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pharmacology
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Transfection
5.Effects of AMPK on the transcriptional activity of FOXO1 and ubiquitin ligase MuRF1 expression in rat cardiomyocytes.
Bao-lin CHEN ; Zhao-jun XIONG ; Cheng-xi ZHANG ; Yue-dong MA ; Rong-sen MENG ; Guang-qin CHEN ; Chen LIU ; Yu-gang DONG
Journal of Southern Medical University 2010;30(11):2419-2422
OBJECTIVETo investigate the effects of AICAR on the activity of transcription factor FOXO1 and expression of ubiquitin ligase MuRF1 in rat cardiomyocytes, and explore the possible role of AMP-activated protein kinase (AMPK) in proteolysis pathways.
METHODSIn vitro cultured neonatal rat cardiac myocytes were treated with AICAR, and Western blotting was used to detect the phosphorylation of FOXO1 and expression of MuRF1 in the cells.
RESULTSAICAR activated AMPK in rat cardiac myocytes. Activated AMPK significantly inhibited the phosphorylation of FOXO1 and increased MuRF1 protein expression.
CONCLUSIONAMPK may regulate proteolysis by activating FOXO1 transcription factor and up-regulating MuRF1 expression.
AMP-Activated Protein Kinases ; metabolism ; Aminoimidazole Carboxamide ; analogs & derivatives ; pharmacology ; Animals ; Cells, Cultured ; Forkhead Transcription Factors ; metabolism ; Muscle Proteins ; metabolism ; Myocytes, Cardiac ; metabolism ; Nerve Tissue Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Ribonucleotides ; pharmacology ; Tripartite Motif Proteins ; Ubiquitin-Protein Ligases ; metabolism
6.PI3K activity is associated with expression of neural specific genes in mouse fetal liver cells enhanced by butylated hydroxyanisole.
Ge-Xiu LIU ; Yuan ZHANG ; Dong-Mei HE
Chinese Journal of Applied Physiology 2006;22(2):237-240
AIMTo study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver cells in vitro.
METHODS14.5-day-old mouse fetal liver-derived cells were cultured, and were induced by 200 micromol/L butylated hydroxyanisole (BHA) combined with PI3K inhibitor LY294002 (20 micromol/L), and then were incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting or RT-PCR.
RESULTSThere was low level of neurofilament-L (NF-L) and brain factor-1 (BF-1) but no neurofilament-H (NF-H) and tyrosine hydroxylase (TH) in fetal liver cells. BHA promoted significantly expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver cells. NF-L mRNA increased 5.8 fold, NF-H mRNA 8.0 fold, BF-1 mRNA 2.68 fold, and TH mRNA 30 fold, respectively (all P < 0.01 vs untreated cells). NF-L protein increased 11.29 fold, NF-H 5.5 fold, BF-1 2.53 fold, TH 4.76 fold. Moreover, expression of these BHA-induced genes were inhibited by PI3K inhibitor LY294002.
CONCLUSIONBHA induced neural differentiation of fetal liver cells through PI3K.
Animals ; Butylated Hydroxyanisole ; pharmacology ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Hepatocytes ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Phosphatidylinositol 3-Kinases ; metabolism
7.microRNA-218 Inhibits Oxygen-induced Retinal Neovascularization via Reducing the Expression of Roundabout 1.
Shuang HAN ; Yi-Chun KONG ; Bei SUN ; Quan-Hong HAN ; Ying CHEN ; Yu-Chuan WANG
Chinese Medical Journal 2016;129(6):709-715
BACKGROUNDThe mechanisms of pathological retinal neovascularization (RNV) remain unknown. Several microRNAs were reported to be involved in the process of RNV. Oxygen-induced retinopathy (OIR) is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 (miR-218) in oxygen-induced RNV.
METHODSOIR was used to establish RNV model. The expression level of miR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 (Robo1) expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay.
RESULTSIn OIR mice, the expression level of miR-218 was significantly down-regulated (P = 0.006). Retinal Robo1 expression was significantly increased at both mRNA and protein levels (P = 0.001, 0.008; respectively). miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robo1.
CONCLUSIONSOur experiments showed that restoration of miR-218 inhibited retinal angiogenesis via targeting Robo1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.
Animals ; Cell Movement ; Cells, Cultured ; Mice ; Mice, Inbred C57BL ; MicroRNAs ; physiology ; Nerve Tissue Proteins ; physiology ; Oxygen ; pharmacology ; Receptors, Immunologic ; physiology ; Retinal Neovascularization ; prevention & control
8.Progress on Correlation between the Expression of CDK5 and Brain Injury Time.
Shi-yu MA ; Ru-bo LI ; LUO YU-JIA ; Meng-yan LÜ ; Han-zhi WANG ; Zheng-yin WANG
Journal of Forensic Medicine 2016;32(1):58-60
Cyclin-dependent kinase 5 (CDK5) is a member of cyclin-dependent kinase family, which does not directly regulate cell cycle. Through phosphorylation of target protein, CDK5 plays an irreplaceable role in the development, reparation and degeneration of neurons. Brain injury refers to the organic injury of brain tissue caused by external force hit on the head. Owing to the stress and repair system activated by our body itself after injury, various proteins and enzymes of the brain tissues are changed quantitatively, which can be used as indicators for estimating the time of injury. This review summarizes the progress on the distribution, the activity mechanism and the physiological effects of CDK5 after brain injury and its corresponding potential served as a marker for brain injury determination.
Brain/physiopathology*
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Brain Injuries/physiopathology*
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Cyclin-Dependent Kinase 5/metabolism*
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Nerve Tissue Proteins/metabolism*
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Neurons
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Neuroprotective Agents/pharmacology*
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Phosphorylation/drug effects*
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Time Factors
9.Intrathecal Lamotrigine Attenuates Antinociceptive Morphine Tolerance and Suppresses Spinal Glial Cell Activation in Morphine-Tolerant Rats.
In Gu JUN ; Sung Hoon KIM ; Yang In YOON ; Jong Yeon PARK
Journal of Korean Medical Science 2013;28(2):300-307
Glial cells play a critical role in morphine tolerance, resulting from repeated administration of morphine. Both the development and the expression of tolerance are suppressed by the analgesic lamotrigine. This study investigated the relationship between the ability of lamotrigine to maintain the antinociceptive effect of morphine during tolerance development and glial cell activation in the spinal cord. In a rat model, morphine (15 microg) was intrathecally injected once daily for 7 days to induce morphine tolerance. Lamotrigine (200 microg) was co-administered with morphine either for 7 days or the first or last 3 days of this 7 day period. Thermal nociception was measured. OX-42 and GFAP immunoreactivity, indicating spinal microglial and astrocytic activation were evaluated on day 8. Tolerance developed after 7 days of intrathecal morphine administration; however, this was completely blocked and reversed by co-administration of lamotrigine. When lamotrigine was coinjected with morphine on days 5-7, the morphine effect was partially restored. Glial cell activation increased with the development of morphine tolerance but was clearly inhibited in the presence of lamotrigine. These results suggest that, in association with the suppression of spinal glial cell activity, intrathecally coadministered lamotrigine attenuates antinociceptive tolerance to morphine.
Analgesics/*pharmacology
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Animals
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Antigens, CD11b/metabolism
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Astrocytes/cytology
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Drug Tolerance
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Immunohistochemistry
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Male
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Microglia/cytology
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Morphine/*pharmacology
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Nerve Tissue Proteins/metabolism
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Neuroglia/cytology/*metabolism
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Rats
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Rats, Sprague-Dawley
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Spinal Cord/*cytology
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Triazines/*pharmacology
10.Gingerol activates noxious cold ion channel TRPA1 in gastrointestinal tract.
Meng-Qi YANG ; Lin-Lan YE ; Xiao-Ling LIU ; Xiao-Ming QI ; Jia-Di LV ; Gang WANG ; Ulah-Khan FARHAN ; Nawaz WAQAS ; Ding-Ding CHEN ; Lei HAN ; Xiao-Hui ZHOU
Chinese Journal of Natural Medicines (English Ed.) 2016;14(6):434-440
TRPA1 channels are non-selective cation channels that could be activated by plant-derived pungent products, including gingerol, a main active constituent of ginger. Ginger could improve the digestive function; however whether ginger improves the digestive function through activating TRPA1 receptor in gastrointestinal tract has not been investigated. In the present study, gingerol was used to stimulate cell lines (RIN14B or STC-1) while depletion of extracellular calcium. TRPA1 inhibitor (rethenium red) and TRPA1 gene silencing via TRPA1-specific siRNA were also used for mechanistic studies. The intracellular calcium and secretion of serotonin or cholecystokinin were measured by fura-2/AM and ELISA. Stimulation of those cells with gingerol increased intracellular calcium levels and the serotonin or cholecystokinin secretion. The gingerol-induced intracellular calcium increase and secretion (serotonin or cholecystokinin) release were completely blocked by ruthenium red, EGTA, and TRPA1-specific siRNA. In summary, our results suggested that gingerol derived from ginger might improve the digestive function through secretion releasing from endocrine cells of the gut by inducing TRPA1-mediated calcium influx.
Calcium
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metabolism
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Calcium Channels
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genetics
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metabolism
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Catechols
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pharmacology
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Cell Line
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Fatty Alcohols
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pharmacology
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Gastrointestinal Tract
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drug effects
;
metabolism
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Ginger
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chemistry
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Humans
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Nerve Tissue Proteins
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genetics
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metabolism
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Plant Extracts
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pharmacology
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TRPA1 Cation Channel
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Transient Receptor Potential Channels
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genetics
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metabolism