Evaluación de los factores determinantes del recuento de plaquetas en pacientes con cirrosis

Thrombocytopenia is considered one of the hallmarks of patients with cirrhosis. Several mechanisms have been implicated in the pathophysiology of thrombocytopenia in cirrhosis. Hypersplenism caused by splenomegay, classically regarded as an indirect marker of portal hypertension has been considered the main factor implicated [200]. Nevertheless, portal hypertension is best estimated by the hepatic venous pressure gradient (HVPG) [32, 189], although contradictory results have been reported regarding the association between HVPG and platelet count [195-197]. The identification of thrombopoietin (TPO), a growth factor that enhances the maturation of megakaryocytes and the release of platelets from the bone marrow, has shed new light on the physiolgy of platelets [217]. In normal conditions in adults, TPO is mainly produced in the liver [93, 96] and the circulating leves of platelets are controlled by a negative feedback mechanism [99], so there is an inverse relationship between the amount of circulating platelets, and the amount of TPO that can reach the bone marrow to stimulate thrombopoiesis. In liver cirrhosis perhaps a decreased syntehesis of TPO could be implicated in the development of thrombocytopenia. Controversial findings regarding the role of each mechanism in thrombocytopenia of liver cirrhosis have been reported [142, 146, 160-161, 184] and no study has simultaneously evaluated the influence of the different mechanisms including portal hypertension and TPO production nor whether their influence could change in different stages of the disease..

The Clinical Significance of DC-SIGN and DC-SIGNR, which Are Novel Markers Expressed in Human Colon Cancer

<div><p>Background</p><p>Colon cancer has always been diagnosed at a late stage, which is associated with poor prognosis. The currently used serum tumor markers CEA and CA19-9 display low sensitivity and specificity and may not have diagnostic value in early stage colon cancer. Thus, there is an urgent need to identify novel serum biomarkers for use in the early detection of colon cancer.</p><p>Methods</p><p>In this study, the expression of DC-SIGN and DC-SIGNR in serum was detected by enzyme-linked immunosorbent assay (ELISA). DC-SIGN and DC-SIGNR expression was detected in cancer tissues by immunohistochemistry (IHC).</p><p>Results</p><p>The level of sDC-SIGN was lower in patients than in the healthy controls, while the level of sDC-SIGNR in patients was higher than in the healthy controls. Both sDC-SIGN and sDC-SIGNR had diagnostic significances for cancer patients, and the combined diagnosis of these two markers was higher than both of them alone. Furthermore, there were significant differences between both sDC-SIGN and sDC-SIGNR in stage I/II patients and the healthy controls. Moreover, high sDC-SIGN level was accompanied with the long survival time. Additionally, DC-SIGNR was negative in the cancer foci and matched normal colon tissues but was weakly positive between the cancer foci. DC-SIGN staining was faint in matched normal colon tissues, strong in the tumor stroma and the invasive margin of colon cancer tissues, and negatively correlated with the sDC-SIGN level in serum from the same patient. Interestingly, the percent survival of patients with a DC-SIGN mean density of>0.001219 (the upper 95% confidence interval of matched normal colon tissues) was higher than for all other patients.</p><p>Conclusion</p><p>DC-SIGN and DC-SIGNR are blood-based molecular markers that can potentially be used for the diagnosis of early stage patients. Moreover, expression of DC-SIGN in serum and cancer tissues may affect the survival time for colon cancer patients.</p></div

Clinical data and mean density of deceased colon cancer patients and live cancer patients in Immuunohistochemical study.

<p>Clinical data and mean density of deceased colon cancer patients and live cancer patients in Immuunohistochemical study.</p

The levels and diagnostic values of sDC-SIGN and sDC-SIGNR in colon cancer patients.

<p><b>A–B</b>: There was statistical significance in the sDC-SIGN and sDC-SIGNR level between healthy controls and colon cancer patients, P<0.001. The sDC-SIGN level (A) was lower in colon cancer patients than healthy controls; however, the sDC-SIGNR level (B) was higher in colon cancer patients than healthy controls. <b>C</b>: The level of sDC-SIGN in serum was significantly correlated with that of sDC-SIGNR in the same patient, P<0.001, r = 0.8137. <b>D–E</b>: According to the analysis of Youden index, the cut-off concentrations for sDC-SIGN and sDC-SIGNR are less than 2.226 µg/ml and more than 227.2 ng/ml, respectively, and the corresponding Sensitivity and Specificity of sDC-SIGN and sDC-SIGNR are 87.56%, 55.56% and 61.90%, 97.50%, respectively. Additionally, the Area under the curve (AUC) for sDC-SIGN is 0.7538, while the AUC of sDC-SIGNR is 0.8031. <b>F–G</b>: The AUC of CEA and CA19-9 in patients were 0.7477 and 0.6126, respectively. Based on the clinical decisive levels (0–5 ug/l for CEA, 0–27 U/ml for CA19-9), the cut-off values of CEA and CA19-9 were obtained. And the corresponding sensitivity and specificity of CEA and CA19-9 were 29.22 and 92.38, 14.67 and 94.55, respectively. <b>H</b>: The combined diagnosis of these two markers, sDC-SIGN and sDC-SIGNR, was analyzed through the binary logistic regression and ROC curve. The AUC of sDC-SIGN/sDC-SIGNR was 0.9885, the specificity and sensitivity were 94.8% and 98.7%, respectively.</p

The early diagnostic values of sDC-SIGN and sDC-SIGNR in stage I/II colon cancer patients.

<p><b>A–B</b>: Both sDC-SIGN and sDC-SIGNR levels from early colon cancer patients. Stage I/II patients were significantly different from healthy people, P<0.01. sDC-SIGN is lower than in the healthy control, while sDC-SIGNR is higher. <b>C–D</b>: In the stage I/II patients, sDC-SIGN and sDC-SIGNR had a significant diagnostic value (P<0.01). The cut-off concentrations of DC-SIGN and DC-SIGNR were less than 2.211 µg/ml and more than 189.3 ng/ml, respectively. The corresponding sensitivity and specificity of the two molecules were 81.33% and 55.56%, 48.65% and 92.50%, respectively. <b>E–F</b>: The AUC of CEA and CA19-9 in early cancer were 0.6904 and 0.6917. At the above clinical decisive level, the sensitivity of both CEA (18.64) and CA19-9 (10.00) were very low, while, the specificity of both CEA (92.33) and CA19-9 (94.55) was high. <b>G</b>: The comparison between the ROC curves of four markers, DC-SIGN, DC-SIGNR, CEA and CA19-9. There were significant differences between the AUCs of sDC-SIGN and both sDC-SIGNR and CA19-9. According to the cut-off values for sDC-SIGN and sDC-SIGNR obtained from the ROC curves, CEA and CA19-9 from the clinical decisive level, the sensitivity (diagnostic values) of both sDC-SIGN (81%) and sDC-SIGNR (49%) were higher than CEA (19%) and CA19-9 (10%).</p

Clinical data of the colon cancer patients in ELISA study and the diagnostic values of different novel markers in the colon cancer with early stage.

<p>Note: “<b>*</b>” means that the samples are from all the colon cancer patients with surgical therapy; “<b>#</b>”means that the samples are from stage ??/?? colon cancer patients with surgical treatment.</p><p>Clinical data of the colon cancer patients in ELISA study and the diagnostic values of different novel markers in the colon cancer with early stage.</p

Semi-quantitative image analysis of DC-SIGN expression in tissues and the analysis of its correlation with sDC-SIGN and CEA in serum from the same patient.

<p><b>A</b>: There was statistical significance in IHC for DC-SIGN expression between colon cancer tissues and matched normal colon tissues, P<0.001. DC-SIGN expression in colon cancer patients was higher than in normal colon tissues; <b>B</b>: The correlation between DC-SIGN staining intensity in colon cancer tissue and sDC-SIGN level in serum from the same patient. The mean density (Y axis) was negatively correlated with sDC-SIGN (X axis), r = −0.4643, P<0.01; <b>C</b>: The correlation between the mean density of DC-SIGN in colon cancer and CEA in serum. No significant correlation was observed between the mean density (Y axis) and CEA (X axis), with a Spearman correlation coefficient of -0.036, P>0.05.</p

Aberrant correlation between DC-SIGN expression in colon cancer tissues and patient survival.

<p><b>A</b>: Survival curves by DC-SIGN expression in colon cancer patients. The percent survival of patients with a mean density>0.001219 was higher than for those with mean density <0.001219 (Mantel-Cox, P = 0.009). <b>B</b>: The mean density of DC-SIGN in stage I/II colon cancer patients were notably lower than that in III or IV colon cancer patients according to TNM staging system, P<0.05.</p

IHC for DC-SIGN and DC-SIGNR expression in colon cancer tissues and in matched normal colonic mucosa of colon cancer patients.

<p>Areas in the black boxes of <b>A</b>, <b>C</b>, <b>E</b>, <b>G</b>, <b>I</b><b>and</b><b><u>A</u></b>, <b><u>C</u></b>, <b><u>E</u></b>, <b><u>G</u></b><u>, <b>I</b></u> were enlarged below. <b>A-B</b>: DC-SIGN expression was detected in the internal border, central and peripheral part of colon carcinoma. <b>C–D</b>: blank control primary human colon cancer sections (without anti-DC-SIGN pAb). <b>E–F</b>: DC-SIGN expression in matched normal colonic mucosa. <b>G–H</b>: DC-SIGN expression was mainly detected in human lymphoid sinus. <b>I–J</b>: blank control human lymph node sections (without anti-DC-SIGN pAb); <b><u>A</u>–<u>B</u></b>: DC-SIGNR expression was weakly detected in the invasive margin of tumor. <b><u>C</u>–<u>D</u></b>: blank control primary human colon cancer sections. <b><u>E</u>–<u>F</u></b>: DC-SIGN expression was negative in matched normal colonic mucosa, while the <b><u>G</u></b> and <b><u>H</u></b> were positive controls in the human lymph node. <b><u>I</u>–<u>J</u></b>: blank control human lymph node sections. Magnification:100× in <b>A</b>, <b>C</b>, <b>E</b>,<u><b>A</b></u>, <b><u>C</u></b>, <b><u>E</u></b>, <b><u>G</u></b>,<u><b>I</b></u>; 200× in <b>G</b>, <b>I</b>; 400× in <b>B</b>, <b>D</b>, <b>F</b>, <b>H</b>, <b>J</b>, <b><u>B</u></b>, <b><u>D</u></b>, <b><u>F</u></b>, <b><u>H</u></b>, <b><u>J</u></b>.</p

Additional file 1: Table S2. of Novel roles of DC-SIGNR in colon cancer cell adhesion, migration, invasion, and liver metastasis

The table shows the clinical data of the colon cancer patients in DC-SIGNR ELISA study. (DOCX 24 kb