Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients
The aim of this research is to explore the effect of miR‐200b‐3p targeting DNMT 3A on the proliferation and apoptosis of osteoarthritis ( OA ) cartilage cells. Quantitative RT ‐ PCR was performed to analyse the expression of miR‐200b‐3p, DNMT 3A , MMP 1 , MMP 3 , MMP 9 , MMP 13 and COL II in normal...
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| Veröffentlicht in: | Journal of cellular and molecular medicine 2017-10, Vol.21 (10), p.2308-2316 |
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| creator | Wu, Jian Tao, Yunjuan Shang, Anquan Wang, Weiwei Zhang, Yujie Hu, Liqing Wang, Jun Wang, Yuan Guo, Naizhou |
| description | The aim of this research is to explore the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of osteoarthritis (
OA
) cartilage cells. Quantitative
RT
‐
PCR
was performed to analyse the expression of miR‐200b‐3p,
DNMT
3A
,
MMP
1
,
MMP
3
,
MMP
9
,
MMP
13
and
COL II
in normal and
OA
cartilage tissues. The dual‐luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR‐200b‐3p and
DNMT
3A
. We also constructed eukaryotic expression vector to overexpress miR‐200b‐3p and
DNMT
3A
. We detected the expression level of
MMP
s
and
COL II
in stable transfected cartilage cells using
RT
‐
PCR
and Western blot. Cell proliferation and apoptosis were evaluated using the
MTS
, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of
OA
cartilage cells. The results of
RT
‐
PCR
indicated that both miR‐200b‐3p and
COL II
were down‐regulated in
OA
cartilage tissues, while the expression of
DNMT
3A
and
MMP
s
was up‐regulated in
OA
cartilage tissues. The expressions of
DNMT
3A
,
MMP
s
and
COL II
detected by Western blot showed the same trend of the results of
RT
‐
PCR
. The dual‐luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR‐200b‐3p and
DNMT
3A
. In overexpressed miR‐200b‐3p cartilage cells,
DNMT
3A
and
MMP
s were significantly down‐regulated,
COL II
was significantly up‐regulated, cell viability was enhanced and apoptosis rate was decreased (
P
< 0.05). In overexpressed
DNM
3T
cartilage cells,
MMP
s
were significantly up‐regulated,
COL II
was significantly down‐regulated, cell viability was weakened and apoptosis rate was increased (
P
< 0.05). MiR‐200b‐3p inhibited the secretion of
MMP
s
, promoted the synthesis of
COL II
and enhanced the growth and proliferation of
OA
cartilage cells through inhibiting the expression of
DNMT
3A
. |
| doi_str_mv | 10.1111/jcmm.13152 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_1943574296</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1943574296</sourcerecordid><originalsourceid>FETCH-LOGICAL-c1046-8335c93f2039ff5856bd70affcd850bdd7fd41885f3198e2b302395c6323f2953</originalsourceid><addsrcrecordid>eNotkM1KAzEUhYMoWKsbnyDgTpia5E6mybLU-gOtgtT1kMkkNnU6GZMUcecj-Iw-iVPbuzmXy3fPgYPQJSUj2s_NWm82IwqUsyM0oFywLJeQHx92KkCcorMY14RAQUEO0PvMWqMT9hanlcGuTSYonZxvcWXSpzEtXriX3-8fRkjVC3RYtTW-fVosMUxwj2kVkmvUm8HaNE3cOfmYjO_Pq-CSi7hTyZk2xXN0YlUTzcVBh-j1bracPmTz5_vH6WSeaUryIhMAXEuwjIC0lgteVPWYKGt1LTip6nps65wKwS1QKQyrgDCQXBfA-ifJYYiu9r5d8B9bE1O59tvQ9pEllTnwcc5k0VPXe0oHH2MwtuyC26jwVVJS7sosd2WW_2XCH1axZ_0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1943574296</pqid></control><display><type>article</type><title>Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients</title><source>Wiley-Blackwell Open Access Titles</source><source>DOAJ Directory of Open Access Journals</source><source>Wiley Online Library Journals Frontfile Complete</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><creator>Wu, Jian ; Tao, Yunjuan ; Shang, Anquan ; Wang, Weiwei ; Zhang, Yujie ; Hu, Liqing ; Wang, Jun ; Wang, Yuan ; Guo, Naizhou</creator><creatorcontrib>Wu, Jian ; Tao, Yunjuan ; Shang, Anquan ; Wang, Weiwei ; Zhang, Yujie ; Hu, Liqing ; Wang, Jun ; Wang, Yuan ; Guo, Naizhou</creatorcontrib><description>The aim of this research is to explore the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of osteoarthritis (
OA
) cartilage cells. Quantitative
RT
‐
PCR
was performed to analyse the expression of miR‐200b‐3p,
DNMT
3A
,
MMP
1
,
MMP
3
,
MMP
9
,
MMP
13
and
COL II
in normal and
OA
cartilage tissues. The dual‐luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR‐200b‐3p and
DNMT
3A
. We also constructed eukaryotic expression vector to overexpress miR‐200b‐3p and
DNMT
3A
. We detected the expression level of
MMP
s
and
COL II
in stable transfected cartilage cells using
RT
‐
PCR
and Western blot. Cell proliferation and apoptosis were evaluated using the
MTS
, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of
OA
cartilage cells. The results of
RT
‐
PCR
indicated that both miR‐200b‐3p and
COL II
were down‐regulated in
OA
cartilage tissues, while the expression of
DNMT
3A
and
MMP
s
was up‐regulated in
OA
cartilage tissues. The expressions of
DNMT
3A
,
MMP
s
and
COL II
detected by Western blot showed the same trend of the results of
RT
‐
PCR
. The dual‐luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR‐200b‐3p and
DNMT
3A
. In overexpressed miR‐200b‐3p cartilage cells,
DNMT
3A
and
MMP
s were significantly down‐regulated,
COL II
was significantly up‐regulated, cell viability was enhanced and apoptosis rate was decreased (
P
< 0.05). In overexpressed
DNM
3T
cartilage cells,
MMP
s
were significantly up‐regulated,
COL II
was significantly down‐regulated, cell viability was weakened and apoptosis rate was increased (
P
< 0.05). MiR‐200b‐3p inhibited the secretion of
MMP
s
, promoted the synthesis of
COL II
and enhanced the growth and proliferation of
OA
cartilage cells through inhibiting the expression of
DNMT
3A
.</description><identifier>ISSN: 1582-1838</identifier><identifier>EISSN: 1582-4934</identifier><identifier>DOI: 10.1111/jcmm.13152</identifier><language>eng</language><publisher>Chichester: John Wiley & Sons, Inc</publisher><subject>Apoptosis ; Arthritis ; Assaying ; Cartilage diseases ; Cell culture ; Cell proliferation ; Collagenase 3 ; Gelatinase B ; Knee ; Osteoarthritis ; Polymerase chain reaction ; Secretion ; Tissues</subject><ispartof>Journal of cellular and molecular medicine, 2017-10, Vol.21 (10), p.2308-2316</ispartof><rights>2017. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c1046-8335c93f2039ff5856bd70affcd850bdd7fd41885f3198e2b302395c6323f2953</citedby><cites>FETCH-LOGICAL-c1046-8335c93f2039ff5856bd70affcd850bdd7fd41885f3198e2b302395c6323f2953</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,860,27901,27902</link.rule.ids></links><search><creatorcontrib>Wu, Jian</creatorcontrib><creatorcontrib>Tao, Yunjuan</creatorcontrib><creatorcontrib>Shang, Anquan</creatorcontrib><creatorcontrib>Wang, Weiwei</creatorcontrib><creatorcontrib>Zhang, Yujie</creatorcontrib><creatorcontrib>Hu, Liqing</creatorcontrib><creatorcontrib>Wang, Jun</creatorcontrib><creatorcontrib>Wang, Yuan</creatorcontrib><creatorcontrib>Guo, Naizhou</creatorcontrib><title>Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients</title><title>Journal of cellular and molecular medicine</title><description>The aim of this research is to explore the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of osteoarthritis (
OA
) cartilage cells. Quantitative
RT
‐
PCR
was performed to analyse the expression of miR‐200b‐3p,
DNMT
3A
,
MMP
1
,
MMP
3
,
MMP
9
,
MMP
13
and
COL II
in normal and
OA
cartilage tissues. The dual‐luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR‐200b‐3p and
DNMT
3A
. We also constructed eukaryotic expression vector to overexpress miR‐200b‐3p and
DNMT
3A
. We detected the expression level of
MMP
s
and
COL II
in stable transfected cartilage cells using
RT
‐
PCR
and Western blot. Cell proliferation and apoptosis were evaluated using the
MTS
, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of
OA
cartilage cells. The results of
RT
‐
PCR
indicated that both miR‐200b‐3p and
COL II
were down‐regulated in
OA
cartilage tissues, while the expression of
DNMT
3A
and
MMP
s
was up‐regulated in
OA
cartilage tissues. The expressions of
DNMT
3A
,
MMP
s
and
COL II
detected by Western blot showed the same trend of the results of
RT
‐
PCR
. The dual‐luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR‐200b‐3p and
DNMT
3A
. In overexpressed miR‐200b‐3p cartilage cells,
DNMT
3A
and
MMP
s were significantly down‐regulated,
COL II
was significantly up‐regulated, cell viability was enhanced and apoptosis rate was decreased (
P
< 0.05). In overexpressed
DNM
3T
cartilage cells,
MMP
s
were significantly up‐regulated,
COL II
was significantly down‐regulated, cell viability was weakened and apoptosis rate was increased (
P
< 0.05). MiR‐200b‐3p inhibited the secretion of
MMP
s
, promoted the synthesis of
COL II
and enhanced the growth and proliferation of
OA
cartilage cells through inhibiting the expression of
DNMT
3A
.</description><subject>Apoptosis</subject><subject>Arthritis</subject><subject>Assaying</subject><subject>Cartilage diseases</subject><subject>Cell culture</subject><subject>Cell proliferation</subject><subject>Collagenase 3</subject><subject>Gelatinase B</subject><subject>Knee</subject><subject>Osteoarthritis</subject><subject>Polymerase chain reaction</subject><subject>Secretion</subject><subject>Tissues</subject><issn>1582-1838</issn><issn>1582-4934</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNotkM1KAzEUhYMoWKsbnyDgTpia5E6mybLU-gOtgtT1kMkkNnU6GZMUcecj-Iw-iVPbuzmXy3fPgYPQJSUj2s_NWm82IwqUsyM0oFywLJeQHx92KkCcorMY14RAQUEO0PvMWqMT9hanlcGuTSYonZxvcWXSpzEtXriX3-8fRkjVC3RYtTW-fVosMUxwj2kVkmvUm8HaNE3cOfmYjO_Pq-CSi7hTyZk2xXN0YlUTzcVBh-j1bracPmTz5_vH6WSeaUryIhMAXEuwjIC0lgteVPWYKGt1LTip6nps65wKwS1QKQyrgDCQXBfA-ifJYYiu9r5d8B9bE1O59tvQ9pEllTnwcc5k0VPXe0oHH2MwtuyC26jwVVJS7sosd2WW_2XCH1axZ_0</recordid><startdate>201710</startdate><enddate>201710</enddate><creator>Wu, Jian</creator><creator>Tao, Yunjuan</creator><creator>Shang, Anquan</creator><creator>Wang, Weiwei</creator><creator>Zhang, Yujie</creator><creator>Hu, Liqing</creator><creator>Wang, Jun</creator><creator>Wang, Yuan</creator><creator>Guo, Naizhou</creator><general>John Wiley & Sons, Inc</general><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QP</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>RC3</scope></search><sort><creationdate>201710</creationdate><title>Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients</title><author>Wu, Jian ; Tao, Yunjuan ; Shang, Anquan ; Wang, Weiwei ; Zhang, Yujie ; Hu, Liqing ; Wang, Jun ; Wang, Yuan ; Guo, Naizhou</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c1046-8335c93f2039ff5856bd70affcd850bdd7fd41885f3198e2b302395c6323f2953</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Apoptosis</topic><topic>Arthritis</topic><topic>Assaying</topic><topic>Cartilage diseases</topic><topic>Cell culture</topic><topic>Cell proliferation</topic><topic>Collagenase 3</topic><topic>Gelatinase B</topic><topic>Knee</topic><topic>Osteoarthritis</topic><topic>Polymerase chain reaction</topic><topic>Secretion</topic><topic>Tissues</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wu, Jian</creatorcontrib><creatorcontrib>Tao, Yunjuan</creatorcontrib><creatorcontrib>Shang, Anquan</creatorcontrib><creatorcontrib>Wang, Weiwei</creatorcontrib><creatorcontrib>Zhang, Yujie</creatorcontrib><creatorcontrib>Hu, Liqing</creatorcontrib><creatorcontrib>Wang, Jun</creatorcontrib><creatorcontrib>Wang, Yuan</creatorcontrib><creatorcontrib>Guo, Naizhou</creatorcontrib><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><jtitle>Journal of cellular and molecular medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wu, Jian</au><au>Tao, Yunjuan</au><au>Shang, Anquan</au><au>Wang, Weiwei</au><au>Zhang, Yujie</au><au>Hu, Liqing</au><au>Wang, Jun</au><au>Wang, Yuan</au><au>Guo, Naizhou</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients</atitle><jtitle>Journal of cellular and molecular medicine</jtitle><date>2017-10</date><risdate>2017</risdate><volume>21</volume><issue>10</issue><spage>2308</spage><epage>2316</epage><pages>2308-2316</pages><issn>1582-1838</issn><eissn>1582-4934</eissn><abstract>The aim of this research is to explore the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of osteoarthritis (
OA
) cartilage cells. Quantitative
RT
‐
PCR
was performed to analyse the expression of miR‐200b‐3p,
DNMT
3A
,
MMP
1
,
MMP
3
,
MMP
9
,
MMP
13
and
COL II
in normal and
OA
cartilage tissues. The dual‐luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR‐200b‐3p and
DNMT
3A
. We also constructed eukaryotic expression vector to overexpress miR‐200b‐3p and
DNMT
3A
. We detected the expression level of
MMP
s
and
COL II
in stable transfected cartilage cells using
RT
‐
PCR
and Western blot. Cell proliferation and apoptosis were evaluated using the
MTS
, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR‐200b‐3p targeting
DNMT
3A
on the proliferation and apoptosis of
OA
cartilage cells. The results of
RT
‐
PCR
indicated that both miR‐200b‐3p and
COL II
were down‐regulated in
OA
cartilage tissues, while the expression of
DNMT
3A
and
MMP
s
was up‐regulated in
OA
cartilage tissues. The expressions of
DNMT
3A
,
MMP
s
and
COL II
detected by Western blot showed the same trend of the results of
RT
‐
PCR
. The dual‐luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR‐200b‐3p and
DNMT
3A
. In overexpressed miR‐200b‐3p cartilage cells,
DNMT
3A
and
MMP
s were significantly down‐regulated,
COL II
was significantly up‐regulated, cell viability was enhanced and apoptosis rate was decreased (
P
< 0.05). In overexpressed
DNM
3T
cartilage cells,
MMP
s
were significantly up‐regulated,
COL II
was significantly down‐regulated, cell viability was weakened and apoptosis rate was increased (
P
< 0.05). MiR‐200b‐3p inhibited the secretion of
MMP
s
, promoted the synthesis of
COL II
and enhanced the growth and proliferation of
OA
cartilage cells through inhibiting the expression of
DNMT
3A
.</abstract><cop>Chichester</cop><pub>John Wiley & Sons, Inc</pub><doi>10.1111/jcmm.13152</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1582-1838 |
| ispartof | Journal of cellular and molecular medicine, 2017-10, Vol.21 (10), p.2308-2316 |
| issn | 1582-1838 1582-4934 |
| language | eng |
| recordid | cdi_proquest_journals_1943574296 |
| source | Wiley-Blackwell Open Access Titles; DOAJ Directory of Open Access Journals; Wiley Online Library Journals Frontfile Complete; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central |
| subjects | Apoptosis Arthritis Assaying Cartilage diseases Cell culture Cell proliferation Collagenase 3 Gelatinase B Knee Osteoarthritis Polymerase chain reaction Secretion Tissues |
| title | Effect of the interaction between MiR‐200b‐3p and DNMT 3A on cartilage cells of osteoarthritis patients |
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