NANOGP8 is the key regulator of stemness, EMT, Wnt pathway, chemoresistance, and other malignant phenotypes in gastric cancer cells
Accumulating evidence demonstrated that NANOG1, the key transcription factor for embryonic stem cells, is associated with human cancers. NANOGP8, one of the pseudogenes in NANOG gene family, contains an intact open reading frame and also said to be expressed in cancer tissues. Therefore, a systemati...
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| description | Accumulating evidence demonstrated that NANOG1, the key transcription factor for embryonic stem cells, is associated with human cancers. NANOGP8, one of the pseudogenes in NANOG gene family, contains an intact open reading frame and also said to be expressed in cancer tissues. Therefore, a systematic study is greatly needed to address the following questions: among NANOG1 and NANOGP8, which gene is the main contributor for NANOG expression in cancer cells and which one is the key regulator responsible for stemness, epithelial-mesenchymal transition (EMT), metastasis, chemoresistance and other malignant phenotypes. Here we try to explore these issues with gastric adenocarcinoma cell lines in vitro using variety of molecular and cellular techniques.
Special primers were designed to distinguish PCR products from NANOG1 and NANOGP8. Sphere-forming cells were cultured with serum-free and selective medium. A stable cell line was established with infection of lentivirus containing NANOGP8. qPCR was performed to measure NANOGP8 expression and its association with stemness, EMT and CSC markers in adherent cells and sphere-forming cells. Western blot analysis was deployed to confirm results of the transcript analysis. Experiments of cell proliferation, migration, invasion, clonogenic assay, sphere cell growth assays, cell cycle analysis, β-catenin accumulation and translocation in nucleus, and drug resistance were conducted to measure the impact of NANOGP8 on malignant statuses of gastric cancer cells. Immunofluorescence staining was used to analyze cell subpopulations with different markers.
NANOGP8 is mainly responsible for NANOG expression in sphere-forming (stem cell-like) cells derived from gastric cancer cell lines regardless their differentiation status. Ectopic expression of NANOGP8 significantly up-regulates stemness transcription factors, EMT inducers, and cancer stem cell markers (CSC) including Lgr5. NANOGP8 also promotes expression of the signature genes vimentin and N-caderin for mesenchymal cells and down-regulates the signature gene E-caderin for epithelial cells whereby confer the cells with mesenchymal cell phenotype. In NANOGP8 over-expressed adherent and sphere-forming cells, Lgr5+ cells are significantly increased. Ectopic expression of NANOGP8 endows gastric cells with enhanced proliferation, migration, invasion, sphere-forming and clonogenic capacity, and chemoresistance. NANOGP8 expression also enhances β-catenin accumulation in nucleus and s |
| doi_str_mv | 10.1371/journal.pone.0192436 |
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Special primers were designed to distinguish PCR products from NANOG1 and NANOGP8. Sphere-forming cells were cultured with serum-free and selective medium. A stable cell line was established with infection of lentivirus containing NANOGP8. qPCR was performed to measure NANOGP8 expression and its association with stemness, EMT and CSC markers in adherent cells and sphere-forming cells. Western blot analysis was deployed to confirm results of the transcript analysis. Experiments of cell proliferation, migration, invasion, clonogenic assay, sphere cell growth assays, cell cycle analysis, β-catenin accumulation and translocation in nucleus, and drug resistance were conducted to measure the impact of NANOGP8 on malignant statuses of gastric cancer cells. Immunofluorescence staining was used to analyze cell subpopulations with different markers.
NANOGP8 is mainly responsible for NANOG expression in sphere-forming (stem cell-like) cells derived from gastric cancer cell lines regardless their differentiation status. Ectopic expression of NANOGP8 significantly up-regulates stemness transcription factors, EMT inducers, and cancer stem cell markers (CSC) including Lgr5. NANOGP8 also promotes expression of the signature genes vimentin and N-caderin for mesenchymal cells and down-regulates the signature gene E-caderin for epithelial cells whereby confer the cells with mesenchymal cell phenotype. In NANOGP8 over-expressed adherent and sphere-forming cells, Lgr5+ cells are significantly increased. Ectopic expression of NANOGP8 endows gastric cells with enhanced proliferation, migration, invasion, sphere-forming and clonogenic capacity, and chemoresistance. NANOGP8 expression also enhances β-catenin accumulation in nucleus and strengthens Wnt signal transduction.
NANOGP8 is the main regulator of gastric cancer stem cells. It is closely associated with EMT, stemness, and CSC marker as well as Wnt signal pathway. NANOGP8 is correlated with cell proliferation, migration, invasion, clonogenic capacity, β-catenin accumulation in nucleus, and chemoresistance in gastric cancer. NANOGP8 is a promising molecular target for clinical intervention of gastric cancer.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0192436</identifier><identifier>PMID: 29689047</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Accumulation ; Adenocarcinoma ; Adherent cells ; Analysis ; Biology and Life Sciences ; Biomarkers ; Biotechnology ; Cancer ; Cell cycle ; Cell differentiation ; Cell growth ; Cell migration ; Cell proliferation ; Chemoresistance ; Colorectal cancer ; Drug resistance ; Ectopic expression ; Education ; Embryo cells ; Embryonic stem cells ; Epithelial cells ; Forming ; Gastric cancer ; Gene expression ; Genetic aspects ; Growth factors ; Immunofluorescence ; Laboratories ; Life sciences ; Medicine and Health Sciences ; Mesenchyme ; Metastases ; Metastasis ; Molecular biology ; Nuclei (cytology) ; Pancreatic cancer ; Penicillin ; Pharmaceutical sciences ; Phenotypes ; Primers ; Pseudogenes ; Research and analysis methods ; Stem cells ; Stomach cancer ; Subpopulations ; Transcription factors ; Transduction ; Translocation ; Tumor cell lines ; Vimentin ; Wnt protein ; β-Catenin</subject><ispartof>PloS one, 2018-04, Vol.13 (4), p.e0192436-e0192436</ispartof><rights>COPYRIGHT 2018 Public Library of Science</rights><rights>2018 Ma et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Ma et al 2018 Ma et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-8c177bbd4b7a1ebc732e136f41b0b04c55f6fee4a1db384088566f792f042bb3</citedby><cites>FETCH-LOGICAL-c692t-8c177bbd4b7a1ebc732e136f41b0b04c55f6fee4a1db384088566f792f042bb3</cites><orcidid>0000-0002-4354-0328</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5915267/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5915267/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29689047$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Katoh, Masaru</contributor><creatorcontrib>Ma, Xia</creatorcontrib><creatorcontrib>Wang, Bei</creatorcontrib><creatorcontrib>Wang, Xiaofang</creatorcontrib><creatorcontrib>Luo, Yujiao</creatorcontrib><creatorcontrib>Fan, Wufang</creatorcontrib><title>NANOGP8 is the key regulator of stemness, EMT, Wnt pathway, chemoresistance, and other malignant phenotypes in gastric cancer cells</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Accumulating evidence demonstrated that NANOG1, the key transcription factor for embryonic stem cells, is associated with human cancers. NANOGP8, one of the pseudogenes in NANOG gene family, contains an intact open reading frame and also said to be expressed in cancer tissues. Therefore, a systematic study is greatly needed to address the following questions: among NANOG1 and NANOGP8, which gene is the main contributor for NANOG expression in cancer cells and which one is the key regulator responsible for stemness, epithelial-mesenchymal transition (EMT), metastasis, chemoresistance and other malignant phenotypes. Here we try to explore these issues with gastric adenocarcinoma cell lines in vitro using variety of molecular and cellular techniques.
Special primers were designed to distinguish PCR products from NANOG1 and NANOGP8. Sphere-forming cells were cultured with serum-free and selective medium. A stable cell line was established with infection of lentivirus containing NANOGP8. qPCR was performed to measure NANOGP8 expression and its association with stemness, EMT and CSC markers in adherent cells and sphere-forming cells. Western blot analysis was deployed to confirm results of the transcript analysis. Experiments of cell proliferation, migration, invasion, clonogenic assay, sphere cell growth assays, cell cycle analysis, β-catenin accumulation and translocation in nucleus, and drug resistance were conducted to measure the impact of NANOGP8 on malignant statuses of gastric cancer cells. Immunofluorescence staining was used to analyze cell subpopulations with different markers.
NANOGP8 is mainly responsible for NANOG expression in sphere-forming (stem cell-like) cells derived from gastric cancer cell lines regardless their differentiation status. Ectopic expression of NANOGP8 significantly up-regulates stemness transcription factors, EMT inducers, and cancer stem cell markers (CSC) including Lgr5. NANOGP8 also promotes expression of the signature genes vimentin and N-caderin for mesenchymal cells and down-regulates the signature gene E-caderin for epithelial cells whereby confer the cells with mesenchymal cell phenotype. In NANOGP8 over-expressed adherent and sphere-forming cells, Lgr5+ cells are significantly increased. Ectopic expression of NANOGP8 endows gastric cells with enhanced proliferation, migration, invasion, sphere-forming and clonogenic capacity, and chemoresistance. NANOGP8 expression also enhances β-catenin accumulation in nucleus and strengthens Wnt signal transduction.
NANOGP8 is the main regulator of gastric cancer stem cells. It is closely associated with EMT, stemness, and CSC marker as well as Wnt signal pathway. NANOGP8 is correlated with cell proliferation, migration, invasion, clonogenic capacity, β-catenin accumulation in nucleus, and chemoresistance in gastric cancer. NANOGP8 is a promising molecular target for clinical intervention of gastric cancer.</description><subject>Accumulation</subject><subject>Adenocarcinoma</subject><subject>Adherent cells</subject><subject>Analysis</subject><subject>Biology and Life Sciences</subject><subject>Biomarkers</subject><subject>Biotechnology</subject><subject>Cancer</subject><subject>Cell cycle</subject><subject>Cell differentiation</subject><subject>Cell growth</subject><subject>Cell migration</subject><subject>Cell proliferation</subject><subject>Chemoresistance</subject><subject>Colorectal cancer</subject><subject>Drug resistance</subject><subject>Ectopic expression</subject><subject>Education</subject><subject>Embryo cells</subject><subject>Embryonic stem cells</subject><subject>Epithelial cells</subject><subject>Forming</subject><subject>Gastric cancer</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Growth factors</subject><subject>Immunofluorescence</subject><subject>Laboratories</subject><subject>Life sciences</subject><subject>Medicine and Health Sciences</subject><subject>Mesenchyme</subject><subject>Metastases</subject><subject>Metastasis</subject><subject>Molecular biology</subject><subject>Nuclei (cytology)</subject><subject>Pancreatic cancer</subject><subject>Penicillin</subject><subject>Pharmaceutical sciences</subject><subject>Phenotypes</subject><subject>Primers</subject><subject>Pseudogenes</subject><subject>Research and analysis methods</subject><subject>Stem cells</subject><subject>Stomach cancer</subject><subject>Subpopulations</subject><subject>Transcription factors</subject><subject>Transduction</subject><subject>Translocation</subject><subject>Tumor cell lines</subject><subject>Vimentin</subject><subject>Wnt protein</subject><subject>β-Catenin</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk1FvFCEUhSdGY2v1HxglMTGa7K4wwzAzLyZNU2uT2hpt9JEAc5mhzsAKjLrP_nFZd9t0TR8MDxD4zgEO3Cx7SvCCFBV5c-Umb8WwWDoLC0yanBbsXrZPmiKfsxwX92-N97JHIVxhXBY1Yw-zvbxhdYNptZ_9Pj88vzj5WCMTUOwBfYMV8tBNg4jOI6dRiDBaCGGGjj9cztBXG9FSxP6nWM2Q6mF0HoIJUVgFMyRsi1yy8WgUg-msWNM9WBdXSwjIWNSJEL1RSK0FHikYhvA4e6DFEODJtj_ILt8dXx69n59dnJweHZ7NFWvyOK8VqSopWyorQUCqqsiBFExTIrHEVJWlZhqACtLKoqa4rkvGdNXkGtNcyuIge76xXQ4u8G18gad4cE5wXZBEnG6I1okrvvRmFH7FnTD874TzHRc-GjUAb1vJJMVMay1oi7UkpS4VLjSVJeRtk7zebneb5AitAhu9GHZMd1es6XnnfvCyIWXOqmTwamvg3fcJQuSjCeu8hAU3bc7dEFxRmtAX_6B3325LdSJdwFjt0r5qbcoPy4LhlBlhiVrcQaXWwmhU-mvapPkdwesdQWIi_IqdmELgp58__T978WWXfXmL7UEMsQ9umKJxNuyCdAMq70LwoG9CJpivS-U6Db4uFb4tlSR7dvuBbkTXtVH8AbAxD0g</recordid><startdate>20180424</startdate><enddate>20180424</enddate><creator>Ma, 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is the key regulator of stemness, EMT, Wnt pathway, chemoresistance, and other malignant phenotypes in gastric cancer cells</title><author>Ma, Xia ; Wang, Bei ; Wang, Xiaofang ; Luo, Yujiao ; Fan, Wufang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-8c177bbd4b7a1ebc732e136f41b0b04c55f6fee4a1db384088566f792f042bb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Accumulation</topic><topic>Adenocarcinoma</topic><topic>Adherent cells</topic><topic>Analysis</topic><topic>Biology and Life Sciences</topic><topic>Biomarkers</topic><topic>Biotechnology</topic><topic>Cancer</topic><topic>Cell cycle</topic><topic>Cell differentiation</topic><topic>Cell growth</topic><topic>Cell migration</topic><topic>Cell proliferation</topic><topic>Chemoresistance</topic><topic>Colorectal cancer</topic><topic>Drug resistance</topic><topic>Ectopic expression</topic><topic>Education</topic><topic>Embryo cells</topic><topic>Embryonic stem cells</topic><topic>Epithelial cells</topic><topic>Forming</topic><topic>Gastric cancer</topic><topic>Gene expression</topic><topic>Genetic aspects</topic><topic>Growth factors</topic><topic>Immunofluorescence</topic><topic>Laboratories</topic><topic>Life sciences</topic><topic>Medicine and Health Sciences</topic><topic>Mesenchyme</topic><topic>Metastases</topic><topic>Metastasis</topic><topic>Molecular biology</topic><topic>Nuclei (cytology)</topic><topic>Pancreatic cancer</topic><topic>Penicillin</topic><topic>Pharmaceutical sciences</topic><topic>Phenotypes</topic><topic>Primers</topic><topic>Pseudogenes</topic><topic>Research and analysis methods</topic><topic>Stem cells</topic><topic>Stomach cancer</topic><topic>Subpopulations</topic><topic>Transcription factors</topic><topic>Transduction</topic><topic>Translocation</topic><topic>Tumor cell lines</topic><topic>Vimentin</topic><topic>Wnt protein</topic><topic>β-Catenin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ma, Xia</creatorcontrib><creatorcontrib>Wang, Bei</creatorcontrib><creatorcontrib>Wang, Xiaofang</creatorcontrib><creatorcontrib>Luo, Yujiao</creatorcontrib><creatorcontrib>Fan, Wufang</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Proquest Nursing & Allied Health Source</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical 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Xiaofang</au><au>Luo, Yujiao</au><au>Fan, Wufang</au><au>Katoh, Masaru</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>NANOGP8 is the key regulator of stemness, EMT, Wnt pathway, chemoresistance, and other malignant phenotypes in gastric cancer cells</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2018-04-24</date><risdate>2018</risdate><volume>13</volume><issue>4</issue><spage>e0192436</spage><epage>e0192436</epage><pages>e0192436-e0192436</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Accumulating evidence demonstrated that NANOG1, the key transcription factor for embryonic stem cells, is associated with human cancers. NANOGP8, one of the pseudogenes in NANOG gene family, contains an intact open reading frame and also said to be expressed in cancer tissues. Therefore, a systematic study is greatly needed to address the following questions: among NANOG1 and NANOGP8, which gene is the main contributor for NANOG expression in cancer cells and which one is the key regulator responsible for stemness, epithelial-mesenchymal transition (EMT), metastasis, chemoresistance and other malignant phenotypes. Here we try to explore these issues with gastric adenocarcinoma cell lines in vitro using variety of molecular and cellular techniques.
Special primers were designed to distinguish PCR products from NANOG1 and NANOGP8. Sphere-forming cells were cultured with serum-free and selective medium. A stable cell line was established with infection of lentivirus containing NANOGP8. qPCR was performed to measure NANOGP8 expression and its association with stemness, EMT and CSC markers in adherent cells and sphere-forming cells. Western blot analysis was deployed to confirm results of the transcript analysis. Experiments of cell proliferation, migration, invasion, clonogenic assay, sphere cell growth assays, cell cycle analysis, β-catenin accumulation and translocation in nucleus, and drug resistance were conducted to measure the impact of NANOGP8 on malignant statuses of gastric cancer cells. Immunofluorescence staining was used to analyze cell subpopulations with different markers.
NANOGP8 is mainly responsible for NANOG expression in sphere-forming (stem cell-like) cells derived from gastric cancer cell lines regardless their differentiation status. Ectopic expression of NANOGP8 significantly up-regulates stemness transcription factors, EMT inducers, and cancer stem cell markers (CSC) including Lgr5. NANOGP8 also promotes expression of the signature genes vimentin and N-caderin for mesenchymal cells and down-regulates the signature gene E-caderin for epithelial cells whereby confer the cells with mesenchymal cell phenotype. In NANOGP8 over-expressed adherent and sphere-forming cells, Lgr5+ cells are significantly increased. Ectopic expression of NANOGP8 endows gastric cells with enhanced proliferation, migration, invasion, sphere-forming and clonogenic capacity, and chemoresistance. NANOGP8 expression also enhances β-catenin accumulation in nucleus and strengthens Wnt signal transduction.
NANOGP8 is the main regulator of gastric cancer stem cells. It is closely associated with EMT, stemness, and CSC marker as well as Wnt signal pathway. NANOGP8 is correlated with cell proliferation, migration, invasion, clonogenic capacity, β-catenin accumulation in nucleus, and chemoresistance in gastric cancer. NANOGP8 is a promising molecular target for clinical intervention of gastric cancer.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>29689047</pmid><doi>10.1371/journal.pone.0192436</doi><tpages>e0192436</tpages><orcidid>https://orcid.org/0000-0002-4354-0328</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2018-04, Vol.13 (4), p.e0192436-e0192436 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2030210831 |
| source | DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Accumulation Adenocarcinoma Adherent cells Analysis Biology and Life Sciences Biomarkers Biotechnology Cancer Cell cycle Cell differentiation Cell growth Cell migration Cell proliferation Chemoresistance Colorectal cancer Drug resistance Ectopic expression Education Embryo cells Embryonic stem cells Epithelial cells Forming Gastric cancer Gene expression Genetic aspects Growth factors Immunofluorescence Laboratories Life sciences Medicine and Health Sciences Mesenchyme Metastases Metastasis Molecular biology Nuclei (cytology) Pancreatic cancer Penicillin Pharmaceutical sciences Phenotypes Primers Pseudogenes Research and analysis methods Stem cells Stomach cancer Subpopulations Transcription factors Transduction Translocation Tumor cell lines Vimentin Wnt protein β-Catenin |
| title | NANOGP8 is the key regulator of stemness, EMT, Wnt pathway, chemoresistance, and other malignant phenotypes in gastric cancer cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-27T10%3A42%3A49IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=NANOGP8%20is%20the%20key%20regulator%20of%20stemness,%20EMT,%20Wnt%20pathway,%20chemoresistance,%20and%20other%20malignant%20phenotypes%20in%20gastric%20cancer%20cells&rft.jtitle=PloS%20one&rft.au=Ma,%20Xia&rft.date=2018-04-24&rft.volume=13&rft.issue=4&rft.spage=e0192436&rft.epage=e0192436&rft.pages=e0192436-e0192436&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0192436&rft_dat=%3Cgale_plos_%3EA536038416%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2030210831&rft_id=info:pmid/29689047&rft_galeid=A536038416&rft_doaj_id=oai_doaj_org_article_ddb6b406fffa4d0fb15f5c03f4b5e2d9&rfr_iscdi=true |