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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" article-type="research-article" dtd-version="1.1" specific-use="sps-1.8" xml:lang="en">
  <front>
    <journal-meta>
      <journal-id journal-id-type="nlm-ta">Arq Bras Cardiol</journal-id>
      <journal-id journal-id-type="publisher-id">abc</journal-id>
      <journal-title-group>
        <journal-title>Arquivos Brasileiros de Cardiologia</journal-title>
        <abbrev-journal-title abbrev-type="publisher">Arq. Bras. Cardiol.</abbrev-journal-title>
      </journal-title-group>
      <issn pub-type="ppub">0066-782X</issn>
      <issn pub-type="epub">1678-4170</issn>
      <publisher>
        <publisher-name>Sociedade Brasileira de Cardiologia - SBC</publisher-name>
      </publisher>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v3">NdZM4GxjFBYLTfLsrzfNn9R</article-id>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v2">S0066-782X2019000600715</article-id>
      <article-id pub-id-type="publisher-id" specific-use="previous-pid">S0066-782X2019005007102</article-id>
      <article-id pub-id-type="doi">10.5935/abc.20190058</article-id>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Original Article</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Clinical Significance of Platelet Volume and Other Platelet Parameters in Acute Myocardial Infarction and Stable Coronary Artery Disease</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0003-4590-599X</contrib-id>
          <name>
            <surname>Ding</surname>
            <given-names>Liumei</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0002-1660-623X</contrib-id>
          <name>
            <surname>Sun</surname>
            <given-names>Lihua</given-names>
          </name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0003-2455-8829</contrib-id>
          <name>
            <surname>Wang</surname>
            <given-names>Feng</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0002-9158-5934</contrib-id>
          <name>
            <surname>Zhu</surname>
            <given-names>Liejun</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0003-2488-9484</contrib-id>
          <name>
            <surname>Zhang</surname>
            <given-names>Ting</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0002-3369-7022</contrib-id>
          <name>
            <surname>Hua</surname>
            <given-names>Fanli</given-names>
          </name>
          <xref ref-type="aff" rid="aff2">2</xref>
          <xref ref-type="corresp" rid="c1"/>
        </contrib>
      </contrib-group>
      <aff id="aff1">
        <label>1</label>
        <institution content-type="orgname">Fudan University</institution>
        <institution content-type="orgdiv1">Jinshan Hospital</institution>
        <institution content-type="orgdiv2">Clinical Laboratory</institution>
        <addr-line>
          <city>Shanghai</city>
        </addr-line>
        <country country="CN">China</country>
        <institution content-type="original">Clinical Laboratory - Jinshan Hospital of Fudan University, Shanghai - China</institution>
      </aff>
      <aff id="aff2">
        <label>2</label>
        <institution content-type="orgname">Zhongshan Hospital</institution>
        <institution content-type="orgdiv1">Department of Hematology</institution>
        <addr-line>
          <city>Shanghai</city>
        </addr-line>
        <country country="CN">China</country>
        <institution content-type="original">Department of Hematology - Zhongshan Hospital, Qingpu Branch, Shanghai - China</institution>
      </aff>
      <author-notes>
        <corresp id="c1"><bold>Mailing Address: Fanli Hua,</bold> N&#176;. 1158 of East Gongyuan Road, Qingpu District. 201700, Shanghai - China. E-mail: <email>huafanli_cn@163.com</email>
				</corresp>
        <fn fn-type="equal">
          <p>Liumei Ding and Lihua Sun contributed equally to this work as the first authorship.</p>
        </fn>
        <fn fn-type="con">
          <p>
            <bold>Author contributions</bold>
          </p>
          <p>Conception and design of the research: Ding L, Hua F; acquisition of data: Ding L, Sun L, Wang F, Zhu L; analysis and interpretation of the data: Ding L, Sun L, Wang F, Zhu L, Zhang T; statistical analysis: Ding L, Sun L, Wang F, Zhang T, Hua F; writing of the manuscript: Ding L, Zhu L, Zhang T, Hua F; critical revision of the manuscript for intellectual content: Hua F. Supervision: Hua F.</p>
        </fn>
        <fn fn-type="conflict">
          <p>
            <bold>Potential Conflict of Interest</bold>
          </p>
          <p>No potential conflict of interest relevant to this article was reported.</p>
        </fn>
      </author-notes>
      <pub-date pub-type="epub">
        <day>01</day>
        <month>04</month>
        <year>2019</year>
      </pub-date>
      <pub-date pub-type="collection">
        <month>06</month>
        <year>2019</year>
      </pub-date>
      <volume>112</volume>
      <issue>6</issue>
      <fpage>715</fpage>
      <lpage>719</lpage>
      <history>
        <date date-type="received">
          <day>21</day>
          <month>05</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>28</day>
          <month>09</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>14</day>
          <month>11</month>
          <year>2018</year>
        </date>
      </history>
      <permissions>
        <license license-type="open-access" xlink:href="https://creativecommons.org/licenses/by/4.0/" xml:lang="en">
          <license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited</license-p>
        </license>
      </permissions>
      <abstract>
        <title>Abstract</title>
        <sec>
          <title>Background:</title>
          <p>Platelets are important in the initiation of thrombosis, and their morphological and functional changes are closely related with the occurrence and development of coronary artery thrombosis. Platelet parameters might be valuable in distinguishing between acute myocardial infarction (AMI) and stable coronary artery disease (SCAD).</p>
        </sec>
        <sec>
          <title>Objective:</title>
          <p>This study was designed to detect and compare changes in platelet parameters, such as mean platelet volume (MPV) in patients with acute myocardial infarction (AMI) and stable coronary artery disease (SCAD) and to investigate their roles in these diseases.</p>
        </sec>
        <sec>
          <title>Methods:</title>
          <p>Specimen collection: Between January 2011 and December 2013, 2 mL of elbow vein blood was drawn from each of 31 patients primarily diagnosed with AMI, 34 SCAD patients and 50 healthy subjects; and placed in EDTA-K2 anticoagulant tubes. Platelet count (PLT), MPV, plateletcrit (PCT), platelet distribution width (PDW), white blood cell (WBC) and neutrophil (NEU) counts were determined using an STKS automated hematology analyzer (Beckman Courter).</p>
        </sec>
        <sec>
          <title>Results:</title>
          <p>Compared with the control group, MPV levels were significantly higher in the AMI and SCAD groups (p &lt; 0.05), while PLT was significantly lower (p &lt; 0.05).</p>
        </sec>
        <sec>
          <title>Conclusion:</title>
          <p>These results suggest that MPV and other related parameters have a certain value in the diagnosis of SCAD and AMI.</p>
        </sec>
      </abstract>
      <kwd-group xml:lang="en">
        <title>Keywords:</title>
        <kwd>Acute Coronary Syndrome/physiopathology</kwd>
        <kwd>Coronary Thrombosis</kwd>
        <kwd>Mean Platelet Volume</kwd>
        <kwd>Myocardial Infarction</kwd>
      </kwd-group>
    </article-meta>
  </front>
  <body>
    <sec sec-type="intro">
      <title>Introduction</title>
      <p>Acute coronary syndromes (ACS) are a group of clinical syndromes, of which pathological bases are coronary atherosclerosis plaques rupture or erosion,<xref ref-type="bibr" rid="B1">1</xref> followed by complete or incomplete occlusive thrombosis. ACS include acute myocardial infarction (AMI) and unstable angina pectoris. Among these, AMI refers to acute focal myocardial necrosis caused by prolonged and severe myocardial ischemia.<xref ref-type="bibr" rid="B2">2</xref>
			</p>
      <p>In 2013, the European Society of Cardiology issued the management guidelines for stable coronary artery disease (SCAD).<xref ref-type="bibr" rid="B3">3</xref> These guidelines clearly pointed out that SCAD also include situations of no symptoms or stable symptoms after the stabilization of acute coronary syndrome, besides stable angina pectoris. These situations cannot be clearly distinguished from ACS. Previous studies have revealed that platelet is an important medium in the initiation of thrombosis, and its morphological and functional changes are closely correlated with the occurrence and development of coronary artery thrombosis.<xref ref-type="bibr" rid="B4">4</xref><sup>,</sup><xref ref-type="bibr" rid="B5">5</xref>
			</p>
      <p>Additionally, mean platelet volume (MPV), one of the platelet parameters in AMI, is significantly higher than in normal subjects, hence MPV is considered as a predictor for AMI.<xref ref-type="bibr" rid="B6">6</xref> However, the value of platelet parameters in distinguishing between AMI and SCAD has not been reported in China. It has been reported that the supersensitive troponin level was elevated in patients with SCAD.<xref ref-type="bibr" rid="B7">7</xref> Although it did not reach the threshold for the diagnosis of myocardial infarction, the prognosis was poorer, when compared with SCAD patients without an elevated troponin level. In our study, differences in platelet parameters such as MPV in AMI patients, SCAD patients and healthy subjects were analyzed, and the significance of these parameters for predicting the disease was investigated.</p>
    </sec>
    <sec sec-type="methods">
      <title>Methods</title>
      <sec>
        <title>General Information</title>
        <p>A total of 31 patients primarily diagnosed with AMI were enrolled in this study. All patients had been diagnosed for the first time, did not undergo anticoagulation or stent percutaneous coronary intervention, and met the international and Chinese guidelines for the diagnosis of AMI. Of these patients, 23 patients were males and eight patients were females; and the mean age of these patients was 64.4 &#177; 11.6 years. Thirty-four patients were diagnosed with SCAD. These patients were required to meet the following criteria: (1) patients diagnosed with AMI for more than two months; (2) patients without pectoralgia; (3) patients who were receiving anticoagulation drug treatment. Of these patients, 28 patients were males and six patients were females; the mean age of these patients was 60.6 &#177; 13.1 years. The control group comprised 50 healthy subjects who underwent physical examinations at the Outpatient Department. Among these subjects, 38 were males and 12 were females, and their mean age was 60.9 &#177; 6.9 years.</p>
        <p>None of the abovementioned patients had severe liver or kidney disease, malignant tumors, idiopathic thrombocytopenia, or thrombocytopenia caused by other primary diseases.</p>
      </sec>
      <sec>
        <title>Sample collection</title>
        <p>In the early morning, 2 mL of elbow vein blood was withdrawn from each of these subjects under a fasting state, without receiving any hemostasis, coagulation and anticoagulation drugs. The collected blood was placed in EDTA-K2 anticoagulant tubes.</p>
      </sec>
      <sec>
        <title>Detection method</title>
        <p>Platelet count (PLT), plateletcrit (PCT), MPV, platelet distribution width (PDW), white blood cell (WBC) and neutrophil (NEU) counts were determined using an STKS automated hematology analyzer (Beckman Courter).</p>
      </sec>
      <sec>
        <title>Statistical analysis</title>
        <p>Statistical analysis was performed using the GraphPad Prism 5 software. Measurement data were presented as mean&#177; standard deviation (x &#177; SD). Comparison of the means in multiple samples was performed using unpaired t-test. p &lt; 0.05 was considered statistically significant. The prediction value of platelet parameters and the total leukocyte count for AMI and SCAD were evaluated using the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC). Correlations between parameters were analyzed using Pearson&#8217;s correlation statistical method.</p>
      </sec>
    </sec>
    <sec sec-type="results">
      <title>Results</title>
      <p>Results of the four platelet parameters and the total WBC count in AMI patients, SCAD patients and normal healthy subjects (<xref ref-type="table" rid="t1">Table 1</xref>).</p>
      <table-wrap id="t1">
        <label>Table 1</label>
        <caption>
          <title>Platelet parameters and total number of white blood cells and neutrophils in the AMI, SCAD and the control groups</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="25%"/>
            <col width="25%"/>
            <col width="25%"/>
            <col width="25%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left">Parameters</th>
              <th align="center">AMI (n = 31)</th>
              <th align="center">SCAD (n = 34)</th>
              <th align="center">control group (n = 50)</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left">PLT (x 10<sup>9</sup>)</td>
              <td align="center">185.84 &#177; 61.26</td>
              <td align="center">193.62 &#177; 47.1</td>
              <td align="center">206.28 &#177; 36.17</td>
            </tr>
            <tr>
              <td align="left">MPV (fL)</td>
              <td align="center">10.42 &#177; 1.26</td>
              <td align="center">9.16 &#177; 1.28</td>
              <td align="center">8.42 &#177; 0.72</td>
            </tr>
            <tr>
              <td align="left">PCT (%)</td>
              <td align="center">0.1777 &#177; 0.0464</td>
              <td align="center">0.1709 &#177; 0.0393</td>
              <td align="center">0.1693 &#177; 0.0287</td>
            </tr>
            <tr>
              <td align="left">PDW (%)</td>
              <td align="center">16.2 &#177; 0.65</td>
              <td align="center">16.0 &#177; 1.37</td>
              <td align="center">16.5 &#177; 0.32</td>
            </tr>
            <tr>
              <td align="left">WBC (x 10<sup>9</sup>/L)</td>
              <td align="center">10.60 &#177; 2.53</td>
              <td align="center">6.61 &#177; 1.54</td>
              <td align="center">6.37 &#177; 1.25</td>
            </tr>
            <tr>
              <td align="left">NEU (x 10<sup>9</sup>/L)</td>
              <td align="center">8.34 &#177; 2.66</td>
              <td align="center">3.99 &#177; 1.25</td>
              <td align="center">3.79 &#177; 0.93</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN1">
            <p>Compared with the control group, the MPV levels were significantly higher (p &lt; 0.01), while the PLT and the PDW were significantly lower (p &lt; 0.05) in the AMI and SCAD groups. Besides, compared to the control group, the WBC and NEU levels were also significantly higher (p &lt; 0.01) in the AMI group, but there was no significant difference in the SCAD group. However, the PCT levels showed no significant difference between the patient groups and the control group.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
      <p>As shown in <xref ref-type="table" rid="t1">Table 1</xref>, compared with the control group, the MPV levels were significantly higher (p &lt; 0.01), while the PLT and the PDW were significantly lower (p &lt; 0.05) in the AMI and SCAD groups. Moreover, compared to the control group, the WBC and NEU levels were also significantly higher (p &lt; 0.01) in the AMI group, but there was no significant difference in the SCAD group. However, the PCT levels had no significant difference between the patient groups and the control group.</p>
      <sec>
        <title>Evaluation of the diagnostic efficacy of each index by the diagnostic test</title>
        <p>As shown in <xref ref-type="fig" rid="f1">Figure 1</xref>, in the AMI group, the AUC of PLT (95% CI) was 0.6474 (0.5206-0.7742) and the p-value compared with controls was &lt; 0.05, which shows that the differences were statistically significant. The AUC of MPV (95%Cl) was 0.9032 (0.8232-0.9832) and the p-value compared with controls was &lt; 0.01, which revealed that the differences were statistically significant. Furthermore, the AUC of PDW (95% CI) was 0.6529 (0.5239-0.7819) and the p-value compared with controls was &lt; 0.05; the AUC of PCT (95% CI) was 0.5687 (0.4364-0.701) and the p-value compared with controls was &lt; 0.05, and the AUC of WBC (95% CI) was 0.9190 (0.8475-0.9906) and the p-value compared with controls was &lt; 0.01. Finally, the AUC of NEU (95% CI) was 0.9310 (0.8678-0.9942) and the p-value compared with controls was &lt; 0.01. These differences were all statistically significant. These results imply that the diagnostic value of PLT, MPV, WBC and NEU was significantly higher in the AMI group.</p>
        <p>
          <fig id="f1">
            <label>Figure 1</label>
            <caption>
              <title>ROC curves of PLT, PDW, MPV, PCT, WBC and NEU for predicting AMI. PLT: platelet count; MPV: mean platelet volume; PCT: plateletcrit; PDW: platelet distribution width; WBC: white blood cell; NEU: neutrophils; AMI: acute myocardial infarction.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-112-06-0715-90058-gf01.jpg"/></fig>
        </p>
        <p>As shown in <xref ref-type="fig" rid="f2">Figure 2</xref>, in the SCAD group, the AUC of PLT (95% CI) was 0.6176 (0.4907-0.7445) and the p-value compared with controls was &gt; 0.05. The AUC of PDW (95% CI) was 0.6818 (0.5554-0.8081) and the p-value compared with controls was &lt; 0.01, with the differences being statistically significant. The AUC of PDW (95% CI) was 0.5609 (0.4268-0.6949) and the p-value compared with controls was &gt;0.05, whereas the AUC of PCT (95% CI) was 0.5332(0.3994-0.6671) and the p-value compared with controls was &gt; 0.05, and the AUC of WBC (95% CI) was 0.5635 (0.4368-0.6903) and the p-value compared with controls was &gt; 0.05. Finally, the AUC of NEU (95% CI) was 0.5447 (0.4138-0.6756) and the p-value compared with controls was &gt; 0.05. These results imply that the diagnostic value of MPV was significantly higher in the SCAD group.</p>
        <p>
          <fig id="f2">
            <label>Figure 2</label>
            <caption>
              <title>ROC curves of PLT, PDW, MPV, PCT, WBC and NEU for predicting SCAD. ROC, receiver operating characteristics; PLT: platelet count; MPV: mean platelet volume; PCT: plateletcrit; PDW: platelet distribution width; WBC: white blood cell; NEU: neutrophils; SCAD: stable coronary artery disease.</title>
            </caption>
            <graphic xlink:href="1678-4170-abc-112-06-0715-90058-gf02.jpg"/></fig>
        </p>
      </sec>
      <sec>
        <title>Correlation analysis of MPV with PLT, PCT and PDW in patients with myocardial infarction</title>
        <p>Pearson&#8217;s correlation coefficient between MPV and PLT was 0.3817; Pearson&#8217;s correlation coefficient between MPV and PCT was 0.1103. Pearson&#8217;s correlation coefficient between MPV and PDW was 0.0726. It indicates that the MPV and PLT in patients with myocardial infarction have a strong correlation.</p>
      </sec>
    </sec>
    <sec sec-type="discussion">
      <title>Discussion</title>
      <p>The key mechanisms of ACS include coronary plaque rupture, platelet aggregation and thrombus formation. Mounting evidence has revealed that inflammation is an important precipitating event in the pathogenesis of ACS.<xref ref-type="bibr" rid="B8">8</xref> Inflammatory response plays an important role in the process of the occurrence, development and prognosis of coronary heart diseases. As markers of inflammatory response, WBC and NEU are closely correlated to cardiovascular system diseases. Studies have revealed that both can directly reflect the level of inflammation in coronary artery lesions, thereby further reflecting the severity of coronary artery lesions.<xref ref-type="bibr" rid="B9">9</xref><sup>,</sup><xref ref-type="bibr" rid="B10">10</xref> Studies have confirmed that the NEU/lymphocyte ratio could serve as a predictor of major adverse cardiovascular events.<xref ref-type="bibr" rid="B11">11</xref><sup>-</sup><xref ref-type="bibr" rid="B13">13</xref> This study revealed that there was a leukocyte-mediated inflammatory response in patients with AMI and that serum leukocyte levels were higher in SCAD and AMI patients than in normal healthy subjects. Furthermore, serum NEU levels were significantly abnormal in AMI patients, suggesting that inflammatory response is more intense in AMI patients. Hence, we consider that the reason may be that the range of inflammation is broader in myocardial infarction.</p>
      <p>MPV reflects the degree of activation of platelets to a certain extent and is considered an important marker of cardiovascular disease.<xref ref-type="bibr" rid="B14">14</xref> Furthermore, it can be used in risk prediction, diagnosis and prognosis assessment of cardiovascular diseases.<xref ref-type="bibr" rid="B15">15</xref><sup>-</sup><xref ref-type="bibr" rid="B17">17</xref>
			</p>
      <p>A recent study revealed that MPV significantly increased in patients with AMI, and the increase in MPV was correlated to the long-term prognosis of AMI patients to a certain extent.<xref ref-type="bibr" rid="B18">18</xref> Our study also revealed that MPV was significantly increased in patients in the AMI and SCAD groups. The increase in MPV is likely correlated to the intensity of inflammatory response in the body.</p>
      <p>MPV has important significance in the evaluation of arterial thrombotic diseases such as cardiovascular thrombotic diseases. Therefore, MPV can act as an independent factor for the diagnosis and assessment of conditions and the curative effect of ACS. MPV levels in both the AMI and SCAD groups were significantly higher than normal levels, while PLT levels were significantly lower. These indicators are worthwhile in practice for disease diagnosis. In addition, based on the assessment of the diagnostic efficacy of the ROC curve for these parameters, the predicting value of MPV and PLT in AMI was superior to that in SCAD. In summary, the present study revealed that MPV- and PLT-related indicators are valuable for AMI and SCAD diagnostic prediction. However, there are two shortcomings of our study: one is that the study design is cross-sectional, because of the difficulty in keeping contact with these patients; the other is that there are only about 30 patients in the AMI and SCAD groups. There is no doubt that these two problems constitute limitations of our study. Further studies need to be carried out in a larger cohorts of patients with AMI and SCAD, with a more appropriate design to confirm its value as a diagnostic marker.</p>
    </sec>
  </body>
  <back>
    <fn-group>
      <fn fn-type="financial-disclosure">
        <p>
          <bold>Sources of Funding</bold>
        </p>
        <p>There were no external funding sources for this study.</p>
      </fn>
      <fn fn-type="other">
        <p>
          <bold>Study Association</bold>
        </p>
        <p>This study is not associated with any thesis or dissertation work.</p>
      </fn>
      <fn fn-type="other">
        <p>
          <bold>Ethics approval and consent to participate</bold>
        </p>
        <p>This study was approved by the Ethics Committee of the Jinshan Hospital, Fudan University under protocol number 785. All the procedures in this study were in accordance with the 1975 Helsinki Declaration, updated in 2013. Informed consent was obtained from all participants included in the study.</p>
      </fn>
    </fn-group>
    <ref-list>
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      <aff id="aff4">
        <label>2</label>
        <institution content-type="original">Department of Hematology - Zhongshan Hospital, Qingpu Branch, Shanghai - China</institution>
      </aff>
      <author-notes>
        <corresp id="c2"><bold>Correspond&#234;ncia: Fanli Hua,</bold> N&#176;. 1158 of East Gongyuan Road, Qingpu District. 201700, Shanghai - China. E-mail: <email>huafanli_cn@163.com</email>
				</corresp>
        <fn fn-type="equal">
          <p>Liumei Ding e Lihua Sun contribu&#237;ram igualmente para este trabalho.</p>
        </fn>
        <fn fn-type="con">
          <p>
            <bold>Contribui&#231;&#227;o dos autores</bold>
          </p>
          <p>Concep&#231;&#227;o e desenho da pesquisa: Ding L, Hua F; Obten&#231;&#227;o de dados: Ding L, Sun L, Wang F, Zhu L; An&#225;lise e interpreta&#231;&#227;o dos dados: Ding L, Sun L, Wang F, Zhu L, Zhang T; An&#225;lise estat&#237;stica: Ding L, Sun L, Wang F, Zhang T, Hua F; Reda&#231;&#227;o do manuscrito: Ding L, Zhu L, Zhang T, Hua F; Revis&#227;o cr&#237;tica do manuscrito quanto ao conte&#250;do intelectual importante: Hua F.</p>
        </fn>
        <fn fn-type="conflict">
          <p>
            <bold>Potencial conflito de interesses</bold>
          </p>
          <p>Declaro n&#227;o haver conflito de interesses pertinentes.</p>
        </fn>
      </author-notes>
      <abstract>
        <title>Resumo</title>
        <sec>
          <title>Fundamento:</title>
          <p>As plaquetas s&#227;o importantes no in&#237;cio da trombose e suas altera&#231;&#245;es morfol&#243;gicas e funcionais est&#227;o intimamente relacionadas com a ocorr&#234;ncia e o desenvolvimento de trombose da art&#233;ria coron&#225;ria. Os par&#226;metros plaquet&#225;rios podem ser valiosos na distin&#231;&#227;o entre infarto agudo do mioc&#225;rdio (IAM) e doen&#231;a arterial coronariana est&#225;vel (DACE).</p>
        </sec>
        <sec>
          <title>Objetivo:</title>
          <p>O objetivo desse estudo foi detectar e comparar altera&#231;&#245;es nos par&#226;metros plaquet&#225;rios, como o volume plaquet&#225;rio m&#233;dio (VPM) em pacientes com infarto agudo do mioc&#225;rdio (IAM) e doen&#231;a arterial coronariana est&#225;vel (DACE) e investigar seu papel nessas doen&#231;as.</p>
        </sec>
        <sec>
          <title>M&#233;todos:</title>
          <p>Coleta de amostras: Entre janeiro de 2011 e dezembro de 2013, foram retirados 2 mL de sangue da veia do antebra&#231;o de cada um dos 31 pacientes diagnosticados principalmente com IAM, 34 pacientes com DACE e 50 indiv&#237;duos saud&#225;veis; e colocado em tubos com anticoagulante EDTA-K2. As contagens de plaquetas (PQT), VPM, massa total de plaquetas (MTP), Amplitude de Distribui&#231;&#227;o de Plaquetas (PDW, do ingl&#234;s <italic>platelet distribution width)</italic>, contagem de gl&#243;bulos brancos (WBC, do ingl&#234;s <italic>white blood cells</italic>) e neutr&#243;filos (NEU) foram determinadas utilizando-se um analisador de hematologia automatizado STKS (Beckman Courter).</p>
        </sec>
        <sec>
          <title>Resultados:</title>
          <p>Comparado com o grupo controle, os n&#237;veis de VPM foram significativamente maiores nos grupos IAM e DACE (p &lt; 0,05), enquanto os n&#237;veis de PQT foram significativamente menores (p &lt; 0,05).</p>
        </sec>
        <sec>
          <title>Conclus&#227;o:</title>
          <p>Esses resultados sugerem que o VPM e outros par&#226;metros associados t&#234;m um certo valor no diagn&#243;stico de DACE e IAM.</p>
        </sec>
      </abstract>
      <kwd-group xml:lang="pt">
        <title>Palavras-chave:</title>
        <kwd>S&#237;ndrome Coronariana Aguda/fisiopatologia</kwd>
        <kwd>Trombose Coron&#225;ria</kwd>
        <kwd>Volume Plaquet&#225;rio M&#233;dio</kwd>
        <kwd>Infarto do Mioc&#225;rdio</kwd>
      </kwd-group>
    </front-stub>
    <body>
      <sec sec-type="intro">
        <title>Introdu&#231;&#227;o</title>
        <p>As s&#237;ndromes coronarianas agudas (SCA) s&#227;o um grupo de s&#237;ndromes cl&#237;nicas, das quais as bases patol&#243;gicas s&#227;o a ruptura ou a eros&#227;o da placa ateroscler&#243;tica coronariana,<xref ref-type="bibr" rid="B1">1</xref> seguidas de trombose oclusiva completa ou incompleta. As SCA incluem infarto agudo do mioc&#225;rdio (IAM) e angina inst&#225;vel. Dentre esses, o IAM refere-se &#224; necrose mioc&#225;rdica focal aguda causada por isquemia mioc&#225;rdica prolongada e grave.<xref ref-type="bibr" rid="B2">2</xref>
				</p>
        <p>Em 2013, a Sociedade Europeia de Cardiologia emitiu as diretrizes de tratamento para doen&#231;a arterial coronariana est&#225;vel (DACE).<xref ref-type="bibr" rid="B3">3</xref> Essas diretrizes indicaram claramente que a DACE tamb&#233;m inclui situa&#231;&#245;es de aus&#234;ncia de sintomas ou sintomas est&#225;veis ap&#243;s a estabiliza&#231;&#227;o da s&#237;ndrome coronariana aguda, al&#233;m de angina est&#225;vel.</p>
        <p>Essas situa&#231;&#245;es n&#227;o podem ser claramente diferenciadas da SCA. Estudos anteriores revelaram que a plaqueta &#233; um meio importante no in&#237;cio da trombose, e suas altera&#231;&#245;es morfol&#243;gicas e funcionais est&#227;o intimamente correlacionadas com a ocorr&#234;ncia e o desenvolvimento de trombose da art&#233;ria coron&#225;ria.<xref ref-type="bibr" rid="B4">4</xref><sup>,</sup><xref ref-type="bibr" rid="B5">5</xref>
				</p>
        <p>Al&#233;m disso, o volume plaquet&#225;rio m&#233;dio (VPM), um dos par&#226;metros plaquet&#225;rios no IAM, &#233; significativamente maior do que em indiv&#237;duos normais, portanto, o VPM &#233; considerado um preditor para IAM.<xref ref-type="bibr" rid="B6">6</xref> Entretanto, o valor dos par&#226;metros plaquet&#225;rios na distin&#231;&#227;o entre IAM e DACE ainda n&#227;o foi relatado na China.</p>
        <p>Foi relatado que o n&#237;vel de troponina ultrassens&#237;vel estava elevado em pacientes com DACE.<xref ref-type="bibr" rid="B7">7</xref> Embora n&#227;o tenha atingido o limiar para o diagn&#243;stico de infarto do mioc&#225;rdio, o progn&#243;stico foi pior, quando comparado com pacientes com DACE sem um n&#237;vel elevado de troponina. Em nosso estudo, foram analisadas diferen&#231;as nos par&#226;metros plaquet&#225;rios, como o VPM em pacientes com IAM, pacientes com diagn&#243;stico de DACE e indiv&#237;duos saud&#225;veis, e investigou-se a signific&#226;ncia desses par&#226;metros para predizer a doen&#231;a.</p>
      </sec>
      <sec sec-type="methods">
        <title>M&#233;todos</title>
        <sec>
          <title>Informa&#231;&#245;es Gerais</title>
          <p>Um total de 31 pacientes diagnosticados principalmente com IAM foram inclu&#237;dos neste estudo. Todos os pacientes foram diagnosticados pela primeira vez, n&#227;o foram submetidos a anti-coagula&#231;&#227;o ou implante de stent por interven&#231;&#227;o coron&#225;ria percut&#226;nea, e atenderam &#224;s diretrizes internacionais e chinesas para o diagn&#243;stico de IAM. Desses pacientes, 23 eram do sexo masculino e oito do sexo feminino; e a m&#233;dia de idade desses pacientes foi de 64,4 &#177; 11,6 anos.</p>
          <p>Trinta e quatro pacientes foram diagnosticados com DACE. Esses pacientes tinham que preencher os seguintes crit&#233;rios: (1) pacientes diagnosticados com IAM por mais de dois meses; (2) pacientes sem dor no peito; (3) pacientes que estavam recebendo tratamento medicamentoso com anti-coagula&#231;&#227;o. Desses pacientes, 28 eram do sexo masculino e seis do sexo feminino; a m&#233;dia de idade desses pacientes foi de 60,6 &#177; 13,1 anos.</p>
          <p>O grupo controle foi composto por 50 indiv&#237;duos saud&#225;veis que foram submetidos a exames f&#237;sicos no ambulat&#243;rio. Entre esses indiv&#237;duos, 38 eram do sexo masculino e 12 do sexo feminino, com m&#233;dia de idade de 60,9 &#177; 6,9 anos.</p>
          <p>Nenhum dos pacientes acima mencionados tinha doen&#231;a hep&#225;tica ou renal grave, tumores malignos, trombocitopenia idiop&#225;tica ou trombocitopenia causada por outras doen&#231;as prim&#225;rias.</p>
        </sec>
        <sec>
          <title>Coleta de amostras</title>
          <p>No in&#237;cio da manh&#227;, retirou-se 2 mL de sangue da veia do antebra&#231;o de cada um desses indiv&#237;duos em estado de jejum, sem receber nenhum medicamento para hemostasia, coagula&#231;&#227;o e anti-coagula&#231;&#227;o. O sangue coletado foi colocado em tubos contendo o anticoagulante EDTA-K2.</p>
        </sec>
        <sec>
          <title>M&#233;todo de detec&#231;&#227;o</title>
          <p>As contagens de plaquetas (PQT), massa total de plaquetas (MTP), VPM, amplitude de distribui&#231;&#227;o de plaquetas (PDW, do ingl&#234;s <italic>platelet distribution width</italic>), gl&#243;bulos brancos (WBC, do ingl&#234;s <italic>white blood cells)</italic> e neutr&#243;filos (NEU) foram determinadas utilizando-se um analisador de hematologia automatizado STKS (Beckman Courter).</p>
        </sec>
        <sec>
          <title>An&#225;lise estat&#237;stica</title>
          <p>A an&#225;lise estat&#237;stica foi realizada utilizando-se o software GraphPad Prism 5. Os dados das medidas foram apresentados como m&#233;dia &#177; desvio padr&#227;o (x &#177; SD). A compara&#231;&#227;o das m&#233;dias em m&#250;ltiplas amostras foi realizada utilizando-se o teste <italic>t</italic> n&#227;o pareado. Um valor de p &lt; 0,05 foi considerado estatisticamente significativo. O valor de predi&#231;&#227;o dos par&#226;metros plaquet&#225;rios e a contagem total de gl&#243;bulos brancos para IAM e DACE foram avaliados utilizando-se a curva ROC (<italic>Receiver Operating Characteristic</italic>) e a &#225;rea sob a curva ROC (AUC). As correla&#231;&#245;es entre os par&#226;metros foram analisadas pelo m&#233;todo estat&#237;stico de correla&#231;&#227;o de Pearson.</p>
        </sec>
      </sec>
      <sec sec-type="results">
        <title>Resultados</title>
        <p>Resultados dos quatro par&#226;metros plaquet&#225;rios e da contagem total de gl&#243;bulos brancos em pacientes com IAM, pacientes com DACE e indiv&#237;duos saud&#225;veis normais (<xref ref-type="table" rid="t2">Tabela 1</xref>).</p>
        <table-wrap id="t2">
          <label>Tabela 1</label>
          <caption>
            <title>Par&#226;metros plaquet&#225;rios e n&#250;mero total de gl&#243;bulos brancos e neutr&#243;filos nos grupos IAM, DACE e controle</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="25%"/>
              <col width="25%"/>
              <col width="25%"/>
              <col width="25%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left">Par&#226;metros</th>
                <th align="center">IAM (n = 31)</th>
                <th align="center">DACE (n = 34)</th>
                <th align="center">Grupo controle (n = 50)</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left">PQT (x 10<sup>9</sup>)</td>
                <td align="center">185,84 &#177; 61,26</td>
                <td align="center">193,62 &#177; 47,1</td>
                <td align="center">206,28 &#177; 36,17</td>
              </tr>
              <tr>
                <td align="left">VPM (fL)</td>
                <td align="center">10,42 &#177; 1,26</td>
                <td align="center">9,16 &#177; 1,28</td>
                <td align="center">8,42 &#177; 0,72</td>
              </tr>
              <tr>
                <td align="left">MTP (%)</td>
                <td align="center">0,1777 &#177; 0,0464</td>
                <td align="center">0,1709 &#177; 0,0393</td>
                <td align="center">0,1693 &#177; 0,0287</td>
              </tr>
              <tr>
                <td align="left">PDW (%)</td>
                <td align="center">16,2 &#177; 0,65</td>
                <td align="center">16,0 &#177; 1,37</td>
                <td align="center">16,5 &#177; 0,32</td>
              </tr>
              <tr>
                <td align="left">WBC (x 10<sup>9</sup>/L)</td>
                <td align="center">10,60 &#177; 2,53</td>
                <td align="center">6,61 &#177; 1,54</td>
                <td align="center">6,37 &#177; 1,25</td>
              </tr>
              <tr>
                <td align="left">NEU (x 10<sup>9</sup>/L)</td>
                <td align="center">8,34 &#177; 2,66</td>
                <td align="center">3,99 &#177; 1,25</td>
                <td align="center">3,79 &#177; 0,93</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN2">
              <p>Comparado com o grupo controle, os n&#237;veis de VPM foram significativamente maiores (p &lt; 0,01), enquanto os n&#237;veis de PQT e a PDW foram significativamente menores (p &lt; 0,05) nos grupos IAM e DACE. Al&#233;m disso, em compara&#231;&#227;o ao grupo controle, os n&#237;veis de gl&#243;bulos brancos e NEU tamb&#233;m foram significativamente maiores (p &lt; 0,01) no grupo IAM, mas n&#227;o houve diferen&#231;a significativa no grupo DACE. No entanto, os n&#237;veis da MTP n&#227;o apresentaram diferen&#231;a significativa entre os grupos de pacientes e o grupo controle. IAM: infarto agudo do mioc&#225;rdio; DACE: doen&#231;a arterial coronariana est&#225;vel; PQT: contagem de plaquetas; VPM: volume de plaquetas m&#233;dio; MTP: massa total de plaquetas; PDW: amplitude de distribui&#231;&#227;o de plaquetas; WBC: gl&#243;bulos brancos; NEU: neutr&#243;filos.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <p>Como mostrado na <xref ref-type="table" rid="t2">Tabela 1</xref>, em compara&#231;&#227;o com o grupo controle, os n&#237;veis do VPM foram significativamente maiores (p &lt; 0,01), enquanto os n&#237;veis de PQT e PDW foram significativamente menores (p &lt; 0,05) nos grupos AMI e DACE. Al&#233;m disso, em compara&#231;&#227;o ao grupo controle, os n&#237;veis de WBC e NEU tamb&#233;m foram significativamente maiores (p &lt; 0,01) no grupo IAM, mas n&#227;o houve diferen&#231;a significativa em compara&#231;&#227;o com o grupo SCAD. No entanto, os n&#237;veis de MTP n&#227;o apresentaram diferen&#231;a significativa entre os grupos de pacientes e o grupo controle.</p>
        <sec>
          <title>Avalia&#231;&#227;o da efic&#225;cia diagn&#243;stica de cada &#237;ndice pelo teste diagn&#243;stico</title>
          <p>Como mostrado na <xref ref-type="fig" rid="f3">Figura 1</xref>, no grupo IAM, a AUC de PQT (IC95%) foi 0,6474 (0,5206-0,7742) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,05, o que mostra que as diferen&#231;as foram estatisticamente significativas. A AUC do VPM (lC95%) foi de 0,9032 (0,8232-0,9832) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,01, o que mostrou que as diferen&#231;as foram estatisticamente significativas. Al&#233;m disso, a AUC da PDW (IC95%) foi 0,6529 (0,5239-0,7819) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,05; a AUC dos WBC (IC95%) foi de 0,5687 (0,4364-0,701) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,05, enquanto a AUC dos WBC (IC95%) foi 0,9190 (0,8475-0,9996) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,01. Finalmente, a AUC dos NEU (IC95%) foi de 0,9310 (0,8678-0,9942) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,01. Essas diferen&#231;as foram todas estatisticamente significativas. Estes resultados mostram que os valores diagn&#243;sticos de PQT, VPM, WBC e NEU foram significativamente maiores no grupo IAM.</p>
          <p>
            <fig id="f3">
              <label>Figura 1</label>
              <caption>
                <title>Curvas ROC de PQT, PDW, VPM, MTP, WBC e NEU para predizer o IAM. ROC: receiver operating characteristics; PQT: contagem de plaquetas; PDW: amplitude de distribui&#231;&#227;o de plaquetas; VPM: volume de plaquetas m&#233;dio; MTP: massa total de plaquetas; WBC: gl&#243;bulos brancos; NEU: neutr&#243;filos; IAM: infarto agudo do mioc&#225;rdio.</title>
              </caption>
              <graphic xlink:href="1678-4170-abc-112-06-0715-90058-gf01-pt.jpg"/></fig>
          </p>
          <p>Como mostrado na <xref ref-type="fig" rid="f4">Figura 2</xref>, no grupo DACE, a AUC dos n&#237;veis de PQT (IC95%) foi de 0,6176 (0,4907-0,7445) e o valor de p em compara&#231;&#227;o com os controles foi &gt; 0,05. A AUC de PDW (IC95%) foi 0,6818 (0,5554-0,8081) e o valor de p em compara&#231;&#227;o com os controles foi &lt; 0,01, sendo as diferen&#231;as estatisticamente significativas. A AUC da PDW (IC95%) foi de 0,5609 (0,4268-0,6949) e o valor de p em compara&#231;&#227;o com os controles foi &gt; 0,05, enquanto que a AUC da MTP (IC95%) foi de 0,5332 (0,3994-0,6671) e o valor p em compara&#231;&#227;o com os controles foi &gt;0,05, enquanto a AUC dos gl&#243;bulos brancos (IC95%) foi de 0,5635 (0,4368-0,6903) e o valor de p em compara&#231;&#227;o com os controles foi &gt; 0,05. Finalmente, a AUC dos n&#237;veis de NEU (IC95%) foi de 0,5447 (0,4138-0,6756) e o valor de p comparado com os controles foi &gt; 0,05. Esses resultados mostram que o valor diagn&#243;stico do VPM foi significativamente mais alto no grupo DACE.</p>
          <p>
            <fig id="f4">
              <label>Figura 2</label>
              <caption>
                <title>Curvas ROC de PQT, PDW, VPM, MTP, WBC e NEU para predizer DACE. ROC: receiver operating characteristics; PQT: contagem de plaquetas; VPM: volume de plaquetas m&#233;dio; MTP: massa total de plaquetas; PDW: amplitude de distribui&#231;&#227;o de plaquetas; WBC: gl&#243;bulos brancos; NEU: neutr&#243;filos; DACE: doen&#231;a arterial coronariana est&#225;vel.</title>
              </caption>
              <graphic xlink:href="1678-4170-abc-112-06-0715-90058-gf02-pt.jpg"/></fig>
          </p>
        </sec>
        <sec>
          <title>An&#225;lise de correla&#231;&#227;o do VPM com PQT, MTP e PDW em pacientes com infarto do mioc&#225;rdio</title>
          <p>O coeficiente de correla&#231;&#227;o de Pearson entre VPM e PQT foi de 0,3817; o coeficiente de correla&#231;&#227;o de Pearson entre VPM e PQT foi de 0,1103. O coeficiente de correla&#231;&#227;o de Pearson entre VPM e PDW foi de 0,0726. Isso indica que o VPM e PQT em pacientes com infarto do mioc&#225;rdio t&#234;m uma forte correla&#231;&#227;o.</p>
        </sec>
      </sec>
      <sec sec-type="discussion">
        <title>Discuss&#227;o</title>
        <p>Os principais mecanismos da SCA incluem ruptura da placa coronariana, agrega&#231;&#227;o plaquet&#225;ria e forma&#231;&#227;o de trombo. Evid&#234;ncias crescentes demonstraram que a inflama&#231;&#227;o &#233; um evento precipitante importante na patog&#234;nese da SCA.<xref ref-type="bibr" rid="B8">8</xref> A resposta inflamat&#243;ria desempenha um papel importante no processo de ocorr&#234;ncia, desenvolvimento e progn&#243;stico de doen&#231;as coronarianas. Como marcadores da resposta inflamat&#243;ria, os n&#237;veis de WBC e o NEU est&#227;o intimamente correlacionados com as doen&#231;as do sistema cardiovascular. Estudos revelaram que ambos podem refletir diretamente o n&#237;vel de inflama&#231;&#227;o nas les&#245;es das art&#233;rias coron&#225;rias, refletindo ainda mais a gravidade das les&#245;es da art&#233;ria coron&#225;ria.<xref ref-type="bibr" rid="B9">9</xref><sup>,</sup><xref ref-type="bibr" rid="B10">10</xref> Estudos confirmaram que a rela&#231;&#227;o NEU / linf&#243;citos poderia servir como preditor de eventos cardiovasculares adversos maiores.<xref ref-type="bibr" rid="B11">11</xref><sup>-</sup><xref ref-type="bibr" rid="B13">13</xref> Esse estudo revelou que havia uma resposta inflamat&#243;ria mediada por gl&#243;bulos brancos em pacientes com IAM e que os n&#237;veis s&#233;ricos dos gl&#243;bulos brancos eram mais altos em pacientes com DACE e IAM do que em indiv&#237;duos saud&#225;veis normais. Al&#233;m disso, os n&#237;veis s&#233;ricos de NEU foram significativamente anormais em pacientes com IAM, sugerindo que a resposta inflamat&#243;ria &#233; mais intensa em pacientes com a doen&#231;a. Assim, consideramos que isso pode ser devido ao fato de que a extens&#227;o da inflama&#231;&#227;o &#233; mais ampla no infarto do mioc&#225;rdio.</p>
        <p>O VPM reflete o grau de ativa&#231;&#227;o das plaquetas at&#233; certo ponto, e &#233; considerado um importante marcador de doen&#231;a cardiovascular.<xref ref-type="bibr" rid="B14">14</xref> Al&#233;m disso, pode ser utilizado na predi&#231;&#227;o de risco, diagn&#243;stico e avalia&#231;&#227;o progn&#243;stica de doen&#231;as cardiovasculares.<xref ref-type="bibr" rid="B15">15</xref><sup>-</sup><xref ref-type="bibr" rid="B17">17</xref>
				</p>
        <p>Um estudo recente revelou que o VPM aumentou significativamente em pacientes com IAM, e o aumento do VPM foi correlacionado com o progn&#243;stico a longo prazo de pacientes com IAM at&#233; certo ponto.<xref ref-type="bibr" rid="B18">18</xref> Nosso estudo tamb&#233;m mostrou que o VPM estava significativamente aumentado em pacientes nos grupos IAM e DACE. O aumento do VPM est&#225; provavelmente correlacionado com a intensidade da resposta inflamat&#243;ria no indiv&#237;duo.</p>
        <p>O VPM tem um significado importante na avalia&#231;&#227;o de doen&#231;as tromb&#243;ticas arteriais, como as doen&#231;as tromb&#243;ticas cardiovasculares. Sendo assim, o VPM pode atuar como um fator independente para o diagn&#243;stico e avalia&#231;&#227;o das condi&#231;&#245;es e o efeito curativo da SCA. Os n&#237;veis de VPM nos grupos IAM e DACE foram significativamente maiores do que os n&#237;veis normais, enquanto os n&#237;veis de PQT foram significativamente menores. Estes indicadores s&#227;o importantes na pr&#225;tica cl&#237;nica para o diagn&#243;stico de doen&#231;as. Al&#233;m disso, com base na avalia&#231;&#227;o da efic&#225;cia diagn&#243;stica da curva ROC para esses par&#226;metros, o valor preditivo de VPM e PQT no IAM foi superior ao valor encontrado no grupo de DACE. Em resumo, o presente estudo revelou que os indicadores relacionados a VPM e PQT s&#227;o valiosos para predizer o diagn&#243;stico de IAM e DACE.</p>
        <p>Entretanto, nosso estudo tem duas falhas: uma &#233; o fato de que o desenho do estudo &#233; transversal, devido &#224; dificuldade em manter contato com esses pacientes; o outro &#233; que existem apenas cerca de 30 pacientes nos grupos IAM e DACE. N&#227;o h&#225; d&#250;vida de que esses dois problemas constituem limita&#231;&#245;es do nosso estudo. Novos estudos precisam ser realizados em uma coorte maior de pacientes com IAM e DACE, com um desenho mais apropriado para confirmar seu valor como marcador diagn&#243;stico.</p>
      </sec>
    </body>
    <back>
      <fn-group>
        <fn fn-type="financial-disclosure">
          <p>
            <bold>Fontes de financiamento</bold>
          </p>
          <p>O presente estudo n&#227;o teve fontes de financiamento externas.</p>
        </fn>
        <fn fn-type="other">
          <p>
            <bold>Vincula&#231;&#227;o acad&#234;mica</bold>
          </p>
          <p>N&#227;o h&#225; vincula&#231;&#227;o deste estudo a programas de p&#243;s-gradua&#231;&#227;o.</p>
        </fn>
        <fn fn-type="other">
          <p>
            <bold>Aprova&#231;&#227;o &#233;tica e consentimento informado</bold>
          </p>
          <p>Este estudo foi aprovado pelo Comit&#234; de &#201;tica do Jinshan Hospital, Fudan University sob o n&#250;mero de protocolo 785. Todos os procedimentos envolvidos nesse estudo est&#227;o de acordo com a Declara&#231;&#227;o de Helsinki de 1975, atualizada em 2013. O consentimento informado foi obtido de todos os participantes inclu&#237;dos no estudo.</p>
        </fn>
      </fn-group>
    </back>
  </sub-article>
</article>
