The Prognostic Significance of White Blood Cell and Platelet Count for Inhospital Mortality and Pneumonia in Acute Ischemic Stroke

Background: We investigated the combined effect of white blood cell (WBC) and platelet count on in-hospital mortality and pneumonia in acute ischemic stroke (AIS) patients. Methods: A total of 3,265 AIS patients enrolled from December 2013 to May 2014 across 22 hospitals in Suzhou city were included in the present study. We divided patients into four groups according to their level of WBC and platelet count: LWHP (low WBC and high platelet), LWLP (low WBC and low platelet), HWHP (high WBC and high platelet), and HWLP (high WBC and low platelet). A logistic regression model was used to estimate the combined effect of WBC and platelet counts on all-cause in-hospital mortality and pneumonia in AIS patients. Results: HWLP was associated with a 2.07-fold increase in the risk of in-hospital mortality in comparison to LWHP (adjusted odds ratio (OR) 2.07; 95% confidence interval (CI), 1.02-4.18; P-trend =0.020). The risk of pneumonia was significantly higher in patients with HWLP than those with LWHP (adjusted OR 2.29; 95% CI, 1.57-3.35; P-trend &lt;0.001). The C-statistic for the combined WBC and platelet count was higher than WBC count or platelet count alone for the prediction of in- -hospital mortality and pneumonia (all P &lt; 0.01). Conclusion: High WBC count combined with a low platelet count level at admission was independently associated with in-hospital mortality and pneumonia in AIS patients. Moreover, the combination of WBC count and platelet count level appeared to be a better predictor than WBC count or platelet count alone. </jats:sec

Synthesis of copper (II) porphyrin complexes and their interaction with c-myc G-quadruplex DNA

Objective: To synthesize copper (II) porphyrin complexes and study their interactions with c-myc G-quadruplex DNA. Methods: The 5-p-hydroxyphenyl-10, 15, 20-tris (p-methoxyphenyl) copper (II) porphyrin complex [p-HTMOPPCu (II)] was synthesized by the conventional heating method. Ultraviolet (UV) titration, fluorescence titration, fluorescence resonance energy transfer (FRET) melting point and competitive assays were used to study the interactions between p-HTMOPPCu (II) and c-myc G-quadruplex DNA. Results: The UV absorption spectrum and fluorescence spectroscopy results indicated that the complex of p-HTMOPPCu (II) bound better with c-myc G-quadruplex DNA; the FRET melting point assay, and competitive melting point assay demonstrated that p-HTMOPPCu (II) could selectively bind and stabilize c-myc G-quadruplex DNA. Conclusion: p-HTMOPPCu (II) can bind and stabilize c-myc G-quadruplex DNA and will potentially be developed into a class of small molecule inhibitors targeting c-myc G-quadruplex DNA for clinical applications in the treatment of tumors

Megakaryocyte- and Platelet-Derived Microparticles as Novel Diagnostic and Prognostic Biomarkers for Immune Thrombocytopenia

Altered cell-derived microparticles (MPs) have been reported in multiple autoimmune diseases. However, the roles of megakaryocyte- and platelet-derived MPs (MKMPs and PMPs) in immune thrombocytopenia (ITP) have not been investigated. In this study, we examined plasma MKMP and PMP levels in patients with ITP and evaluated their potential diagnostic values. Plasma MKMP and PMP levels were analyzed by flow cytometry in a discovery set of ITP patients (n = 78), non-immune thrombocytopenia (TP) patients (n = 69), and age- and gender-matched healthy controls (n = 88). Samples from a therapy set of ITP patients (n = 21) were used to assess the response to thrombopoietin receptor agonist (TPO-RA) treatment. Spearman correlation analysis was performed between MP levels and disease parameters. Receiver operator characteristic (ROC) curves were generated to evaluate the diagnostic values of the MPs. We found that plasma MKMP and PMP levels were significantly lower in ITP patients than those in healthy controls (p values p p p values p r = 0.558, p < 0.01). Our results indicate that plasma MKMP and PMP levels are decreased in ITP patients and that plasma MKMP and PMP levels may serve as biomarkers for ITP diagnosis and prediction of TPO-RA treatment response

Megakaryocyte- and Platelet-Derived Microparticles as Novel Diagnostic and Prognostic Biomarkers for Immune Thrombocytopenia

Altered cell-derived microparticles (MPs) have been reported in multiple autoimmune diseases. However, the roles of megakaryocyte- and platelet-derived MPs (MKMPs and PMPs) in immune thrombocytopenia (ITP) have not been investigated. In this study, we examined plasma MKMP and PMP levels in patients with ITP and evaluated their potential diagnostic values. Plasma MKMP and PMP levels were analyzed by flow cytometry in a discovery set of ITP patients (n = 78), non-immune thrombocytopenia (TP) patients (n = 69), and age- and gender-matched healthy controls (n = 88). Samples from a therapy set of ITP patients (n = 21) were used to assess the response to thrombopoietin receptor agonist (TPO-RA) treatment. Spearman correlation analysis was performed between MP levels and disease parameters. Receiver operator characteristic (ROC) curves were generated to evaluate the diagnostic values of the MPs. We found that plasma MKMP and PMP levels were significantly lower in ITP patients than those in healthy controls (p values &lt; 0.0001) but higher than in those in TP patients (p &lt; 0.002 and p &lt; 0.0002, respectively). After normalization to platelet counts, PMP/Platelet ratios in ITP patients were higher than those in TP patients and healthy controls (p values &lt; 0.001). PMP/Platelet ratios had a diagnostic value for ITP (area under the curve = 0.808, p &lt; 0.0001) with 73.1% sensitivity and 77.3% specificity. MKMP levels can be used to discriminate ITP from TP with a cut-off value of 112.5 MPs/&mu;L and a sensitivity of 74.4%. Moreover, both MKMP and PMP levels were elevated in ITP patients who responded to TPO-RA treatment. Plasma PMP levels positively correlated with platelet counts in the responders (r = 0.558, p &lt; 0.01). Our results indicate that plasma MKMP and PMP levels are decreased in ITP patients and that plasma MKMP and PMP levels may serve as biomarkers for ITP diagnosis and prediction of TPO-RA treatment response

Combined Effect of White Blood Cell and Platelet Count for Predicting In-Hospital Mortality and Pneumonia in Acute Ischemic Stroke

Abstract Background: High white blood cell (WBC) count was the risk factors for mortality and pneumonia after acute ischemic stroke (AIS). Low platelet count increased the risk of mortality. We investigated the combined effect of WBC count and platelet count on hospital admission and in-hospital mortality and pneumonia in acute AIS patients.Methods: A total of 3,265 AIS patients enrolled from December 2013 to May 2014 across 22 hospitals in Suzhou city were included in the present study. We divided patients into four groups according to their level of WBC count and platelet count: LWHP (low WBC and high platelet), LWLP (low WBC and low platelet), HWHP (high WBC and high platelet) and HWLP (high WBC and low platelet). Cox proportional logistic regression model were used to estimate the combined effect of WBC count and platelet count on all cause in-hospital mortality and pneumonia in AIS patients. Results: HWLP was associated with a 2.07-fold increase in the risk of in-hospital mortality in comparison to LWHP (adjusted odds ratio [OR] 2.07; 95% confidence interval [CI], 1.02-4.18; P-trend =0.020). The risk of pneumonia was significantly higher in patients with HWLP compared to those with LWHP (adjusted OR 2.29; 95% CI, 1.57-3.35; P-trend &lt;0.001). The C-statistic for the combined WBC count and platelet count was higher than WBC count or platelet count alone for prediction of in-hospital mortality and pneumonia (all p &lt; 0.01). Conclusions: High WBC count combined with low platelet count level at admission was independently associated with in-hospital mortality and pneumonia in AIS patients. Moreover, the combination of WBC count and platelet count level appeared to be a better predictor than WBC count or platelet count alone.</jats:p

Flow cytometric immunobead assay for quantitative detection of platelet autoantibodies in immune thrombocytopenia patients

Abstract Background Platelet autoantibody detection is critical for immune thrombocytopenia (ITP) diagnosis and prognosis. Therefore, we aimed to establish a quantitative flow cytometric immunobead assay (FCIA) for ITP platelet autoantibodies evaluation. Methods Capture microbeads coupled with anti-GPIX, -GPIb, -GPIIb, -GPIIIa and P-selectin antibodies were used to bind the platelet-bound autoantibodies complex generated from plasma samples of 250 ITP patients, 163 non-ITP patients and 243 healthy controls, a fluorescein isothiocyanate (FITC)-conjugated secondary antibody was the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Intra- and inter-assay variations of the quantitative FCIA assay were assessed. Comparisons of the specificity, sensitivity and accuracy between quantitative and qualitative FCIA or monoclonal antibody immobilization of platelet antigen (MAIPA) assay were performed. Finally, treatment process was monitored by our quantitative FCIA in 8 newly diagnosed ITPs. Results The coefficient of variations (CV) of the quantitative FCIA assay were respectively 9.4, 3.8, 5.4, 5.1 and 5.8% for anti-GPIX, -GPIb, -GPIIIa, -GPIIb and -P-selectin autoantibodies. Elevated levels of autoantibodies against platelet glycoproteins GPIX, GPIb, GPIIIa, GPIIb and P-selectin were detected by our quantitative FCIA in ITP patients compared to non-ITP patients or healthy controls. The sensitivity, specificity and accuracy of our quantitative assay were respectively 73.13, 81.98 and 78.65% when combining all 5 autoantibodies, while the sensitivity, specificity and accuracy of MAIPA assay were respectively 41.46, 90.41 and 72.81%. Conclusions A quantitative FCIA assay was established. Reduced levels of platelet autoantibodies could be confirmed by our quantitative FCIA in ITP patients after corticosteroid treatment. Our quantitative assay is not only good for ITP diagnosis but also for ITP treatment monitoring

Plasma microRNAs characterising patients with immune thrombo cytopenic purpura

SummaryAltered microRNA (miRNA) expression has been reported in patients with immune thrombocytopenic purpura (ITP). However, the detailed expression profiling of cell-free circulating miRNAs in ITP patients has not been fully investigated. In this study, we aimed to examine plasma miRNAs in ITP patients and evaluate their diagnostic values. Plasma samples from 74 ITP patients and 58 healthy controls were obtained and allocated into discovery, validation, and therapy-response sets. Initial screen with a miRNA microarray assay identified 23 miRNAs with different levels between ITP patients and healthy controls (&gt;1.5-fold changes; p&lt;0.01). Subsequent quantitative real-time PCR confirmed eight up-regulated miRNAs (miR-320c, miR-642b-3p, miR-1275, miR-3141, miR-4270, miR-4499, miR-4739 and miR-6126) and three down-regulated miRNAs (miR-144–3p, miR-1281 and miR-3162–3p) in ITP patients. The levels of these circulating miRNAs varied, depending on ITP subtypes, i.e. newly-diagnosed, persistent and chronic ITP, and between treatment responders and non-responders. In receiver operator characteristic analysis, 10 miRNAs had positive diagnostic values (p&lt;0.05) when tested individually. The diagnostic value improved when the miRNAs were analysed as a panel or together with the analysis of anti-platelet autoantibodies. Plasma miR-3162–3p levels were also found to positively correlate with platelet counts in ITP patients (r=0.338, p=0.01). Our results indicate that plasma miRNA profiles are altered in ITP patients and that the differentially expressed miRNAs may be used as biomarkers to improve the diagnosis of ITP.</jats:p