Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan
Thrombomodulin ( THBD ) is an endothelial surface glycoprotein receptor, having a pivotal role in maintaining laminar blood flow. It functions to protect endothelial integrity by exhibiting anti-coagulation and anti-inflammatory properties thereby playing a key role in cardiovascular disease (CVD) p...
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| Veröffentlicht in: | Molecular biology reports 2020-10, Vol.47 (10), p.7575-7582 |
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| creator | Rafiq, Muhammad Liaquat, Afrose Saeed, Nadia Shamshad, Ghassan Umair Mumtaz, Sana Khan, Muhammad Jawad |
| description | Thrombomodulin (
THBD
) is an endothelial surface glycoprotein receptor, having a pivotal role in maintaining laminar blood flow. It functions to protect endothelial integrity by exhibiting anti-coagulation and anti-inflammatory properties thereby playing a key role in cardiovascular disease (CVD) pathology. Cholesterol lowering drugs have shown to alter the anti-inflammatory effects of cytokines. Understanding the molecular aspects of
THBD
gene and its relation to inflammatory cytokines is important to identify new prognostic and therapeutic targets for the CVD treatments. The present study was conducted to measure the expression of
THBD
,
TNF
-α and
NF-kB
genes in coronary artery disease patients (CAD) in Pakistani population. Lipid profile and BMI was compared both on fifty CAD patients and fifty healthy individuals. Expression analysis for
THBD
,
TNF
-α and
NF-kB
was carried out using real time PCR. The effect of lipid lowering drugs on cardiometabolic risk variables especially gene expression was analyzed. Our results indicated that the difference in BMI was marginal; however LDL-cholesterol and triglycerides levels in CAD patients were significantly higher than healthy individuals.
THBD
gene was significantly up-regulated whereas
TNF
-α and
NF-kB
were significantly down regulated in CAD individuals. Further exploration revealed that these variations were accounted to the use of statins by the patients. The use of statins by CAD patients up-regulated the mRNA expression of
THBD
by down-regulation of inflammatory mediators. The enhanced expression of endothelial
THBD
in response to cholesterol lowering drugs establishes a novel pleiotropic target that can be of clinical significance in thromboembolic and inflammatory disorders. |
| doi_str_mv | 10.1007/s11033-020-05824-6 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2442845446</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2577748325</sourcerecordid><originalsourceid>FETCH-LOGICAL-c375t-a794bfff9ab543b82e8ed2d1218be6dbfb34fe08bca7744f9a2678000ad813da3</originalsourceid><addsrcrecordid>eNp9kUtOHDEQhq2IKAwkF2ARWWKTBQ5-jt1LQAGiIMiCrC27XQ1Npu3B7pbCsXIRzoSH4SGxYFUl-avPLv8I7TD6nVGq9wtjVAhCOSVUGS7J_AOaMaUFkY02G2hGBWVEGsU20VYpN5RSybT6hDYFbwRthJyh9gQiYPi3zFBKnyJOHR6vcxp8GlKYFn3cw5fnx-T-P3Yx4Nr9OsR9xG3KKbp8h10eoZbQF3AF8NKNPcSxrDy_3d--jC5-Rh87tyjw5aluoz_HPy6PTsnZxcnPo4Mz0gqtRuJ0I33XdY3zSgpvOBgIPDDOjId58J0XsgNqfOu0lrJyfK5NXcoFw0RwYht9W3uXOd1OUEY79KWFxcJFSFOxXEpupJJyXtHdN-hNmnKsr7Nc6ao3gqtK8TXV5lRKhs4ucz_UrS2jdhWBXUdgawT2MQK7Un99Uk9-gPAy8vznFRBroNSjeAX59e53tA_7f5HG</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2577748325</pqid></control><display><type>article</type><title>Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan</title><source>MEDLINE</source><source>SpringerLink Journals - AutoHoldings</source><creator>Rafiq, Muhammad ; Liaquat, Afrose ; Saeed, Nadia ; Shamshad, Ghassan Umair ; Mumtaz, Sana ; Khan, Muhammad Jawad</creator><creatorcontrib>Rafiq, Muhammad ; Liaquat, Afrose ; Saeed, Nadia ; Shamshad, Ghassan Umair ; Mumtaz, Sana ; Khan, Muhammad Jawad</creatorcontrib><description>Thrombomodulin (
THBD
) is an endothelial surface glycoprotein receptor, having a pivotal role in maintaining laminar blood flow. It functions to protect endothelial integrity by exhibiting anti-coagulation and anti-inflammatory properties thereby playing a key role in cardiovascular disease (CVD) pathology. Cholesterol lowering drugs have shown to alter the anti-inflammatory effects of cytokines. Understanding the molecular aspects of
THBD
gene and its relation to inflammatory cytokines is important to identify new prognostic and therapeutic targets for the CVD treatments. The present study was conducted to measure the expression of
THBD
,
TNF
-α and
NF-kB
genes in coronary artery disease patients (CAD) in Pakistani population. Lipid profile and BMI was compared both on fifty CAD patients and fifty healthy individuals. Expression analysis for
THBD
,
TNF
-α and
NF-kB
was carried out using real time PCR. The effect of lipid lowering drugs on cardiometabolic risk variables especially gene expression was analyzed. Our results indicated that the difference in BMI was marginal; however LDL-cholesterol and triglycerides levels in CAD patients were significantly higher than healthy individuals.
THBD
gene was significantly up-regulated whereas
TNF
-α and
NF-kB
were significantly down regulated in CAD individuals. Further exploration revealed that these variations were accounted to the use of statins by the patients. The use of statins by CAD patients up-regulated the mRNA expression of
THBD
by down-regulation of inflammatory mediators. The enhanced expression of endothelial
THBD
in response to cholesterol lowering drugs establishes a novel pleiotropic target that can be of clinical significance in thromboembolic and inflammatory disorders.</description><identifier>ISSN: 0301-4851</identifier><identifier>EISSN: 1573-4978</identifier><identifier>DOI: 10.1007/s11033-020-05824-6</identifier><identifier>PMID: 32930934</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Adult ; Aged ; Animal Anatomy ; Animal Biochemistry ; Anti-inflammatory agents ; Biomedical and Life Sciences ; Blood flow ; Cardiovascular disease ; Cardiovascular diseases ; Cholesterol ; Coronary artery ; Coronary Artery Disease - drug therapy ; Coronary Artery Disease - metabolism ; Coronary Artery Disease - pathology ; Coronary vessels ; Cytokines ; Down-regulation ; Drug development ; Female ; Gene expression ; Gene Expression Regulation ; Heart diseases ; Histology ; Humans ; Inflammatory diseases ; Life Sciences ; Low density lipoprotein ; Male ; Middle Aged ; Morphology ; NF-kappa B - biosynthesis ; NF-κB protein ; Original Article ; Pakistan ; Patients ; Statins ; Thromboembolism ; Thrombomodulin ; Thrombomodulin - biosynthesis ; Triglycerides ; Tumor Necrosis Factor-alpha - biosynthesis ; Tumor necrosis factor-α</subject><ispartof>Molecular biology reports, 2020-10, Vol.47 (10), p.7575-7582</ispartof><rights>Springer Nature B.V. 2020</rights><rights>Springer Nature B.V. 2020.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-a794bfff9ab543b82e8ed2d1218be6dbfb34fe08bca7744f9a2678000ad813da3</citedby><cites>FETCH-LOGICAL-c375t-a794bfff9ab543b82e8ed2d1218be6dbfb34fe08bca7744f9a2678000ad813da3</cites><orcidid>0000-0002-9570-1517 ; 0000-0001-6410-6953</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11033-020-05824-6$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11033-020-05824-6$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32930934$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rafiq, Muhammad</creatorcontrib><creatorcontrib>Liaquat, Afrose</creatorcontrib><creatorcontrib>Saeed, Nadia</creatorcontrib><creatorcontrib>Shamshad, Ghassan Umair</creatorcontrib><creatorcontrib>Mumtaz, Sana</creatorcontrib><creatorcontrib>Khan, Muhammad Jawad</creatorcontrib><title>Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan</title><title>Molecular biology reports</title><addtitle>Mol Biol Rep</addtitle><addtitle>Mol Biol Rep</addtitle><description>Thrombomodulin (
THBD
) is an endothelial surface glycoprotein receptor, having a pivotal role in maintaining laminar blood flow. It functions to protect endothelial integrity by exhibiting anti-coagulation and anti-inflammatory properties thereby playing a key role in cardiovascular disease (CVD) pathology. Cholesterol lowering drugs have shown to alter the anti-inflammatory effects of cytokines. Understanding the molecular aspects of
THBD
gene and its relation to inflammatory cytokines is important to identify new prognostic and therapeutic targets for the CVD treatments. The present study was conducted to measure the expression of
THBD
,
TNF
-α and
NF-kB
genes in coronary artery disease patients (CAD) in Pakistani population. Lipid profile and BMI was compared both on fifty CAD patients and fifty healthy individuals. Expression analysis for
THBD
,
TNF
-α and
NF-kB
was carried out using real time PCR. The effect of lipid lowering drugs on cardiometabolic risk variables especially gene expression was analyzed. Our results indicated that the difference in BMI was marginal; however LDL-cholesterol and triglycerides levels in CAD patients were significantly higher than healthy individuals.
THBD
gene was significantly up-regulated whereas
TNF
-α and
NF-kB
were significantly down regulated in CAD individuals. Further exploration revealed that these variations were accounted to the use of statins by the patients. The use of statins by CAD patients up-regulated the mRNA expression of
THBD
by down-regulation of inflammatory mediators. The enhanced expression of endothelial
THBD
in response to cholesterol lowering drugs establishes a novel pleiotropic target that can be of clinical significance in thromboembolic and inflammatory disorders.</description><subject>Adult</subject><subject>Aged</subject><subject>Animal Anatomy</subject><subject>Animal Biochemistry</subject><subject>Anti-inflammatory agents</subject><subject>Biomedical and Life Sciences</subject><subject>Blood flow</subject><subject>Cardiovascular disease</subject><subject>Cardiovascular diseases</subject><subject>Cholesterol</subject><subject>Coronary artery</subject><subject>Coronary Artery Disease - drug therapy</subject><subject>Coronary Artery Disease - metabolism</subject><subject>Coronary Artery Disease - pathology</subject><subject>Coronary vessels</subject><subject>Cytokines</subject><subject>Down-regulation</subject><subject>Drug development</subject><subject>Female</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Heart diseases</subject><subject>Histology</subject><subject>Humans</subject><subject>Inflammatory diseases</subject><subject>Life Sciences</subject><subject>Low density lipoprotein</subject><subject>Male</subject><subject>Middle Aged</subject><subject>Morphology</subject><subject>NF-kappa B - biosynthesis</subject><subject>NF-κB protein</subject><subject>Original Article</subject><subject>Pakistan</subject><subject>Patients</subject><subject>Statins</subject><subject>Thromboembolism</subject><subject>Thrombomodulin</subject><subject>Thrombomodulin - biosynthesis</subject><subject>Triglycerides</subject><subject>Tumor Necrosis Factor-alpha - biosynthesis</subject><subject>Tumor necrosis factor-α</subject><issn>0301-4851</issn><issn>1573-4978</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kUtOHDEQhq2IKAwkF2ARWWKTBQ5-jt1LQAGiIMiCrC27XQ1Npu3B7pbCsXIRzoSH4SGxYFUl-avPLv8I7TD6nVGq9wtjVAhCOSVUGS7J_AOaMaUFkY02G2hGBWVEGsU20VYpN5RSybT6hDYFbwRthJyh9gQiYPi3zFBKnyJOHR6vcxp8GlKYFn3cw5fnx-T-P3Yx4Nr9OsR9xG3KKbp8h10eoZbQF3AF8NKNPcSxrDy_3d--jC5-Rh87tyjw5aluoz_HPy6PTsnZxcnPo4Mz0gqtRuJ0I33XdY3zSgpvOBgIPDDOjId58J0XsgNqfOu0lrJyfK5NXcoFw0RwYht9W3uXOd1OUEY79KWFxcJFSFOxXEpupJJyXtHdN-hNmnKsr7Nc6ao3gqtK8TXV5lRKhs4ucz_UrS2jdhWBXUdgawT2MQK7Un99Uk9-gPAy8vznFRBroNSjeAX59e53tA_7f5HG</recordid><startdate>20201001</startdate><enddate>20201001</enddate><creator>Rafiq, Muhammad</creator><creator>Liaquat, Afrose</creator><creator>Saeed, Nadia</creator><creator>Shamshad, Ghassan Umair</creator><creator>Mumtaz, Sana</creator><creator>Khan, Muhammad Jawad</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7TK</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</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>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-9570-1517</orcidid><orcidid>https://orcid.org/0000-0001-6410-6953</orcidid></search><sort><creationdate>20201001</creationdate><title>Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan</title><author>Rafiq, Muhammad ; Liaquat, Afrose ; Saeed, Nadia ; Shamshad, Ghassan Umair ; Mumtaz, Sana ; Khan, Muhammad Jawad</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c375t-a794bfff9ab543b82e8ed2d1218be6dbfb34fe08bca7744f9a2678000ad813da3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Adult</topic><topic>Aged</topic><topic>Animal Anatomy</topic><topic>Animal Biochemistry</topic><topic>Anti-inflammatory agents</topic><topic>Biomedical and Life Sciences</topic><topic>Blood flow</topic><topic>Cardiovascular disease</topic><topic>Cardiovascular diseases</topic><topic>Cholesterol</topic><topic>Coronary artery</topic><topic>Coronary Artery Disease - drug therapy</topic><topic>Coronary Artery Disease - metabolism</topic><topic>Coronary Artery Disease - pathology</topic><topic>Coronary vessels</topic><topic>Cytokines</topic><topic>Down-regulation</topic><topic>Drug development</topic><topic>Female</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Heart diseases</topic><topic>Histology</topic><topic>Humans</topic><topic>Inflammatory diseases</topic><topic>Life Sciences</topic><topic>Low density lipoprotein</topic><topic>Male</topic><topic>Middle Aged</topic><topic>Morphology</topic><topic>NF-kappa B - biosynthesis</topic><topic>NF-κB protein</topic><topic>Original Article</topic><topic>Pakistan</topic><topic>Patients</topic><topic>Statins</topic><topic>Thromboembolism</topic><topic>Thrombomodulin</topic><topic>Thrombomodulin - biosynthesis</topic><topic>Triglycerides</topic><topic>Tumor Necrosis Factor-alpha - biosynthesis</topic><topic>Tumor necrosis factor-α</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rafiq, Muhammad</creatorcontrib><creatorcontrib>Liaquat, Afrose</creatorcontrib><creatorcontrib>Saeed, Nadia</creatorcontrib><creatorcontrib>Shamshad, Ghassan Umair</creatorcontrib><creatorcontrib>Mumtaz, Sana</creatorcontrib><creatorcontrib>Khan, Muhammad Jawad</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</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>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><collection>MEDLINE - Academic</collection><jtitle>Molecular biology reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rafiq, Muhammad</au><au>Liaquat, Afrose</au><au>Saeed, Nadia</au><au>Shamshad, Ghassan Umair</au><au>Mumtaz, Sana</au><au>Khan, Muhammad Jawad</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan</atitle><jtitle>Molecular biology reports</jtitle><stitle>Mol Biol Rep</stitle><addtitle>Mol Biol Rep</addtitle><date>2020-10-01</date><risdate>2020</risdate><volume>47</volume><issue>10</issue><spage>7575</spage><epage>7582</epage><pages>7575-7582</pages><issn>0301-4851</issn><eissn>1573-4978</eissn><abstract>Thrombomodulin (
THBD
) is an endothelial surface glycoprotein receptor, having a pivotal role in maintaining laminar blood flow. It functions to protect endothelial integrity by exhibiting anti-coagulation and anti-inflammatory properties thereby playing a key role in cardiovascular disease (CVD) pathology. Cholesterol lowering drugs have shown to alter the anti-inflammatory effects of cytokines. Understanding the molecular aspects of
THBD
gene and its relation to inflammatory cytokines is important to identify new prognostic and therapeutic targets for the CVD treatments. The present study was conducted to measure the expression of
THBD
,
TNF
-α and
NF-kB
genes in coronary artery disease patients (CAD) in Pakistani population. Lipid profile and BMI was compared both on fifty CAD patients and fifty healthy individuals. Expression analysis for
THBD
,
TNF
-α and
NF-kB
was carried out using real time PCR. The effect of lipid lowering drugs on cardiometabolic risk variables especially gene expression was analyzed. Our results indicated that the difference in BMI was marginal; however LDL-cholesterol and triglycerides levels in CAD patients were significantly higher than healthy individuals.
THBD
gene was significantly up-regulated whereas
TNF
-α and
NF-kB
were significantly down regulated in CAD individuals. Further exploration revealed that these variations were accounted to the use of statins by the patients. The use of statins by CAD patients up-regulated the mRNA expression of
THBD
by down-regulation of inflammatory mediators. The enhanced expression of endothelial
THBD
in response to cholesterol lowering drugs establishes a novel pleiotropic target that can be of clinical significance in thromboembolic and inflammatory disorders.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>32930934</pmid><doi>10.1007/s11033-020-05824-6</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0002-9570-1517</orcidid><orcidid>https://orcid.org/0000-0001-6410-6953</orcidid></addata></record> |
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| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | Adult Aged Animal Anatomy Animal Biochemistry Anti-inflammatory agents Biomedical and Life Sciences Blood flow Cardiovascular disease Cardiovascular diseases Cholesterol Coronary artery Coronary Artery Disease - drug therapy Coronary Artery Disease - metabolism Coronary Artery Disease - pathology Coronary vessels Cytokines Down-regulation Drug development Female Gene expression Gene Expression Regulation Heart diseases Histology Humans Inflammatory diseases Life Sciences Low density lipoprotein Male Middle Aged Morphology NF-kappa B - biosynthesis NF-κB protein Original Article Pakistan Patients Statins Thromboembolism Thrombomodulin Thrombomodulin - biosynthesis Triglycerides Tumor Necrosis Factor-alpha - biosynthesis Tumor necrosis factor-α |
| title | Gene expression of thrombomodulin, TNF-α and NF-KB in coronary artery disease patients of Pakistan |
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