1.Research Advances in CKLFSF-like MARVEL Transmembrane Domain Containing Member 3.
Feng-zhan HU ; Zheng-zuo SHENG ; Cai-peng QIN ; Tao XU
Acta Academiae Medicinae Sinicae 2016;38(3):360-363
CKLF-like MARVEL transmembrane domain containing member/chemokine-like factor super family member (CKLFSF/CMTM) is a novel tumor suppressor gene. CMTM3 is broadly expressed in normal human tissues and evolutionary conserved,especially in testis,spleen,and some cells of peripheral blood mononuclear cells. However,its expression is undetectable or down-regulated in most carcinoma cell lines and tissues. Restoration of CMTM3 may inhibit the proliferation,migration,and invasion of carcinoma cells. Although the exact mechanism of its anti-tumor activity remains unclear,CKLFSF3/CMTM3 is closely connected with immune system and associated with sex during tumorigenesis. The study advances of CKLFSF3/CMTM3 are elaborated in this review as CMTM3 may be a new target in the gene therapies for tumors,especially genitourinary tumors,while further studies on CMTM3 and its anti-tumor mechanisms are warranted.
Cell Transformation, Neoplastic
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Chemokines
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genetics
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physiology
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Down-Regulation
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Humans
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Leukocytes, Mononuclear
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MARVEL Domain-Containing Proteins
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genetics
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physiology
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Male
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Neoplasms
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pathology
2.Detection and functional annotation of misregulated microRNAs in the brain of the Ts65Dn mouse model of Down syndrome.
Xiang-jun HE ; Yun XIAO ; Qi ZHANG ; Li-ping MA ; Na LI ; Jing YANG
Chinese Medical Journal 2013;126(1):108-113
BACKGROUNDBrain hypoplasia and mental retardation in Down syndrome (DS) can be attributed to a severe and selective disruption of neurogenesis. Secondary disruption of the transcriptome, as well as primary gene dosage imbalance, is responsible for the phenotype. MicroRNA (miRNA) expression is relatively abundant in brain tissue. Perturbed miRNA expression might contribute to the cellular events underlying the pathology in DS.
METHODSMiRNA expression profiles in the cerebrum of Ts65Dn mice, a DS model, were examined with a real-time RT-PCR array. MiRNA target gene expression was detected by real-time quantitative PCR and Western blotting. Based on the prediction of their cerebrum-specific targets, the functions of the misregulated miRNAs were annotated by Gene Ontology (GO) enrichment analysis.
RESULTSA total of 342 miRNAs were examined. Among them, 20 miRNAs showed decreased expression in the brains of Ts65Dn mice, and some of these belonged to the same family. Two known targets of the miR-200 family, Lfng and Zeb2, were specifically selected to compare their expression in the cerebrum of Ts65Dn mice with those of euploids. However, no significant difference was found in terms of mRNA and protein expression levels of these genes. By enrichment analysis of the cerebrum-specific targets of each miRNA, we found that 15 of the differential miRNAs could significantly affect target genes that were enriched in the GO biological processes related to nervous system development.
CONCLUSIONPerturbed expression of multiple functionally cooperative miRNAs contributes to the cellular events underlying the pathogenesis of DS.
Animals ; Brain ; metabolism ; Disease Models, Animal ; Down Syndrome ; etiology ; genetics ; Gene Expression Regulation ; Mice ; MicroRNAs ; physiology
3.MicroRNA-33b inhibits cell proliferation in hepatocellular carcinoma via targeting SALL4.
Yan LI ; Ronghua LI ; Xiaoyu FU ; Wei ZHOU ; Shifang PENG ; Lei FU
Journal of Central South University(Medical Sciences) 2016;41(9):905-910
OBJECTIVE:
To investigate the expression of miR-33b in hepatocellular carcinoma (HCC) and to explore regulatory mechanism of miR-33b for cell proliferation of HCC.
METHODS:
HCC tissues and adjacent non-tumor tissues were collected for this study (n=32 for each). Real-time PCR and Western blot were conducted to examine the mRNA and protein expression, respectively. MTT assay was used to detect the cell proliferation. Luciferase reporter gene assay was performed to verify the target relationship between miR-33b and Sal-like 4 (SALL4).
RESULTS:
MiR-33b was significantly downregulated in HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-33b decreased the proliferation of HCC LH86 cells. SALL4 was identified as a target gene of miR-33b, and its protein expression was negatively regulated by miR-33b. Overexpression of SALL4 reversed the suppressive effect of miR-33b on LH86 cell proliferation. SALL4 was significantly upregulated in HCC tissues compared with adjacent non-tumor tissues.
CONCLUSION
The miR-33b suppresses HCC cell proliferation through down-regulation of SALL4.
Carcinoma, Hepatocellular
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chemistry
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genetics
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physiopathology
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Cell Proliferation
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genetics
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physiology
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Down-Regulation
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Gene Expression Regulation, Neoplastic
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genetics
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physiology
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Humans
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Liver Neoplasms
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MicroRNAs
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analysis
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genetics
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physiology
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RNA, Messenger
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Real-Time Polymerase Chain Reaction
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Transcription Factors
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genetics
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physiology
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Tumor Cells, Cultured
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Up-Regulation
4.Inhibitory effects of AcSDKP on proliferation of human bone marrow mesenchymal stem cells in vitro.
Guo DAI ; Chang HUANG ; Ye LI ; Yi-Hua PI ; Bao-He WANG
Acta Physiologica Sinica 2006;58(2):110-115
The tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP), a physiological regulator of cell proliferation, has been principally reported as a potent inhibitor of the proliferation of haematopoietic stem cells and progenitors. The purpose of this study was to investigate whether the AcSDKP may directly affect the proliferative potential of human bone marrow mesenchymal stem cells (MSCs) in vitro. We added AcSDKP to the cultures of human bone marrow mononuclear cells and measured the number and average area of MSC colonies. MTT colorimetric assay and mitotic index determination were further used to examine the proliferative state of the third passage MSCs in subcultures with or without the addition of AcSDKP. In addition, we evaluated whether AcSDKP may kill MSCs by the trypan blue dye exclusion test. The results showed that the colony forming capacity, the number of viable cells and the mitotic index were reduced in human bone marrow MSCs cultured in 1x10(-12) mol/L to 1x10(-9) mol/L AcSDKP. Maximum inhibitory activity appeared in 1x10(-11) mol/L of AcSDKP. No difference in percent of living cells was observed between the MSC subcultures with and without the addition of AcSDKP. As a result, AcSDKP within a certain range of concentrations has negatively regulatory effects on the proliferation of human bone marrow MSCs in vitro.
Bone Marrow Cells
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cytology
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Cell Proliferation
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drug effects
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Cells, Cultured
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Down-Regulation
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physiology
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Growth Inhibitors
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physiology
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Humans
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Mesenchymal Stromal Cells
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cytology
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Oligopeptides
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physiology
5.Down-regulation of Nrf2-ARE signaling pathway.
Xiu-jun WANG ; Jia-guo WU ; Xiu-wen TANG
Journal of Zhejiang University. Medical sciences 2010;39(1):1-5
Antioxidants
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metabolism
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pharmacology
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Down-Regulation
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Gene Expression Regulation
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Humans
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NF-E2-Related Factor 2
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genetics
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metabolism
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physiology
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Neoplasms
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genetics
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metabolism
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Oxidative Stress
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genetics
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physiology
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Response Elements
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physiology
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Signal Transduction
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physiology
7.MicroRNA-34a inhibits human brain glioma cell growth by down-regulation of Notch1.
Xiao YU ; Wendi ZHANG ; Qin NING ; Xiaoping LUO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(3):370-374
The effects of microRNA-34a (miR-34a)-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups (P<0.05). Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups (P<0.05), but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.
Cell Line, Tumor
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Cell Proliferation
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Down-Regulation
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genetics
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Glioma
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pathology
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physiopathology
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Humans
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MicroRNAs
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genetics
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Receptor, Notch1
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physiology
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Transfection
8.Effect of the expression of ghrelin receptors on the postoperative underpowered small intestinal motility in rats.
Cheng-guang YANG ; Song YU ; Zhi-gang WANG ; Qi ZHENG
Chinese Journal of Gastrointestinal Surgery 2011;14(6):455-458
OBJECTIVETo investigate the effect of the expression of ghrelin receptors on the postoperative small intestine dysmotility in rat models.
METHODSThe effect of different concentrations of ghrelin (0, 0.01, 0.1, 0.5, 1.0 μmol/L) on the contraction of smooth muscle strips of rat small intestine in the presence or absence of carbachol was observed in vitro. End-to-side anastomosis was performed in the study group and sham controls were used. The expression of ghrelin receptors(GHS-R1a) in small intestine muscle layers was detected by immunohistochemistry and Western blot.
RESULTSIn vitro, ghrelin enhanced the contraction of smooth muscle strips in the presence of carbachol, and the differences in contraction induced by different concentrations of ghrelin(0.1, 0.5, 1.0 μmol/L) were statistically significant [(223±18)%, (245±22)%, (264±25)%, P<0.01]. Immunohistochemistry study showed that GHS-R1a mainly located in the muscular layer of the bowel wall. The expression of GHS-R1a in the circular and longitudinal muscle was significantly weaker than that in the control group. The expression of ghrelin receptors after surgery was down-regulated in the study group, which was lower than that in the control group(0.51±0.02 vs. 0.71±0.01, P<0.01).
CONCLUSIONDown regulation of ghrelin receptors in small intestine muscle layers may contribute to the occurrence of small intestine dysmotility after intestinal surgery.
Animals ; Down-Regulation ; Gastrointestinal Motility ; drug effects ; physiology ; Ghrelin ; pharmacology ; Intestine, Small ; drug effects ; metabolism ; physiology ; surgery ; Male ; Postoperative Period ; Rats ; Rats, Sprague-Dawley ; Receptors, Ghrelin ; metabolism
9.miR-494-3p reduces insulin sensitivity in diabetic cardiomyocytes by down-regulation of insulin receptor substrate 1.
Jie WU ; Xing-Hua QIN ; Zuo-Xu HOU ; Zi-Hao FU ; Guo-Hua LI ; Hong-Yan YANG ; Xing ZHANG ; Feng GAO
Acta Physiologica Sinica 2019;71(2):271-278
More and more evidence suggests that microRNA is widely involved in the regulation of cardiovascular function. Our preliminary experiment showed that miR-494-3p was increased in heart of diabetic rats, and miR-494-3p was reported to be related to metabolism such as obesity and exercise. Therefore, this study was aimed to explore the role of miR-494-3p in diabetic myocardial insulin sensitivity and the related mechanism. The diabetic rat model was induced by high fat diet (45 kcal% fat, 12 weeks) combined with streptozotocin (STZ, 30 mg/kg), and cardiac tissue RNA was extracted for qPCR. The results showed that the level of miR-494-3p was significantly up-regulated in the myocardium of diabetic rats compared with the control (P < 0.05). The level of miR-494-3p in H9c2 cells cultured in high glucose and high fat medium (HGHF) was significantly increased (P < 0.01) with the increase of sodium palmitate concentration, whereas down-regulation of miR-494-3p in HGHF treated cells led to an increase in insulin-stimulated glucose uptake (P < 0.01) and the ratio of p-Akt/Akt (P < 0.05). Over-expression of miR-494-3p in H9c2 cell line significantly inhibited insulin-stimulated glucose uptake and phosphorylation of Akt (P < 0.01). Bioinformatics combined with Western blotting experiments confirmed insulin receptor substrate 1 (IRS1) as a target molecule of miR-494-3p. These results suggest that miR-494-3p reduces insulin sensitivity in diabetic cardiomyocytes by down-regulating IRS1.
Animals
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Diabetes Mellitus, Experimental
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physiopathology
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Down-Regulation
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Insulin
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Insulin Receptor Substrate Proteins
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physiology
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Insulin Resistance
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MicroRNAs
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genetics
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Myocytes, Cardiac
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physiology
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Rats
10.Interleukin-10 Down-Regulates Cathepsin B Expression in Fetal Rat Alveolar Type II Cells Exposed to Hyperoxia.
Yonsei Medical Journal 2013;54(2):445-452
PURPOSE: Hyperoxia has the chief biological effect of cell death. We have previously reported that cathepsin B (CB) is related to fetal alveolar type II cell (FATIIC) death and pretreatment of recombinant IL-10 (rIL-10) attenuates type II cell death during 65%-hyperoixa. In this study, we investigated what kinds of changes of CB expression are induced in FATIICs at different concentrations of hyperoxia (65%- and 85%-hyperoxia) and whether pretreatment with rIL-10 reduces the expression of CB in FATIICs during hyperoxia. MATERIALS AND METHODS: Isolated embryonic day 19 fetal rat alveolar type II cells were cultured and exposed to 65%- and 85%-hyperoxia for 12 h and 24 h. Cells in room air were used as controls. Cytotoxicity was assessed by lactate dehydrogenase (LDH) released into the supernatant. Expression of CB was analyzed by fluorescence-based assay upon cell lysis and western blotting, and LDH-release was re-analyzed after preincubation of cathepsin B-inhibitor (CBI). IL-10 production was analyzed by ELISA, and LDH-release was re-assessed after preincubation with rIL-10 and CB expression was re-analyzed by western blotting and real-time PCR. RESULTS: LDH-release and CB expression in FATIICs were enhanced significantly in an oxygen-concentration-dependent manner during hyperoxia, whereas caspase-3 was not activated. Preincubation of FATIICs with CBI significantly reduced LDH-release during hyperoxia. IL-10-release decreased in an oxygen-concentration-dependent fashion, and preincubation of the cells with rIL-10 significantly reduced cellular necrosis and expression of CB in FATIICs which were exposed to 65%- and 85%-hyperoxia. CONCLUSION: Our study suggests that CB is enhanced in an oxygen-concentration-dependent manner, and IL-10 has an inhibitory effect on CB expression in FATIICs during hyperoxia.
Animals
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Cathepsin B/*genetics/metabolism
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*Down-Regulation
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Gene Expression Regulation
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Hyperoxia/*genetics
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Interleukin-10/*pharmacology/physiology
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L-Lactate Dehydrogenase/metabolism
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Necrosis/chemically induced
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Oxygen/metabolism
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Rats