Clinical Significance of the Balanceshift of Th1 and Th2 Type Cytokines in Patients with Primary Immune Thrombocytopenic Purpura Detected by Cytometric Bead Array / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the levels of Th1/Th2 cytokines in peripheral blood of patients with primary immune thrombocytopenic purpura(ITP) before and after treatment and their clinical significance.</p><p><b>METHODS</b>Ninety-eight cases of ITP were treated with glucocorticoid(GC), then were divided into 2 groups: effectively treated group and uneffectively treated group according to efficacy of treatment, 40 healthy persons confirmed by health examination were selected and enrolled in control group. The levels of Th1 cytokines(IFN-γ,TNFα,IL-2) and Th2 cytokines(IL-4,IL-5,IL-10) were detected by cytometric bead array before and at 4 weeks, 3 and 6 months after treatment and the relationship among detected indexes was analyzed.</p><p><b>RESULTS</b>Before treatment with glucocorticoid, the levels of Th1 type cytokines were in 98 patients with ITP were higher and the levels of Th2 type cytokines were lower, compared with the healthy controls(P<0.05). The IL-2/IL-4 ratio was significantly higher than that of healthy controls(P<0.05). After treatment, the levels of Th1 type cytokines in effectively treated group were significantly decreased and the levels of Th2 type cytokines were significantly increased, compared with level before treatment(P<0.05). The IL-2/IL-4 ratio was significantly decreased after treatment for 6 months, compared with that before treatment(1.05±0.43 vs 2.53±0.72)(P<0.05), but the level of Th1 or Th2 type cytokines did not obviously changed.</p><p><b>CONCLUSION</b>Peripheral blood Th1 and Th2 cells express abnormally in ITP patients, ITP is a Th1 dominated disease; the change of IL-2/IL-4 ratio before and after treatment correlated with the prognosis of ITP patients, displaying clinical significance for ITP individual therapy.</p>

Fas-FasL and caspase-3 signal transduction pathway and apoptosis of peripheral T lymphocytes in ITP patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the relationship between Fas-FasL-mediated signal transduction pathway and apoptosis of T lymphocyte subset in ITP patients.</p><p><b>METHODS</b>The expression rates of membrane Fas, FasL and intracellular activated caspase-3 in peripheral T lymphocyte subset were determined by flow cytometry. T cell subsets with caspase-3 protein expression were detected by Western blot.</p><p><b>RESULTS</b>As compared with that in healthy control group [(29.4 +/- 8.2)%], the expression rate of membrane Fas on CD4+ T cells was significantly increased in ITP patients [(42.1 +/- 9.5)%] (P < 0.05), however, that on CD8+ T cells was only slightly increased [(9.3 +/- 6.0)% vs (13.4 +/- 5.8)%] with no statistical significance (P > 0.05). The expression rate of FasL on T cell subset in ITP patients was significantly increased (P < 0.05), and that of intracellular activated caspase-3 in T cell subset in ITP patients was notably higher than that in healthy control group (P < 0.05). Western blot analysis showed that the expression of pro-caspase-3 and cleaved-caspase-3 in CD4+ T cells in patients with ITP after treatment were significantly reduced compared with those before treatment (P < 0.05).</p><p><b>CONCLUSION</b>Apoptosis of T lymphocyte subset in ITP patients is accelerated. It is possible that Fas-FasL signal transduction pathway plays an important role in the induction of the apoptosis. The degree of apoptosis of T lymphocytes closely correlates with the disease's activity in ITP patients. Hormone therapy may interfere with Fas-FasL signal transduction pathway of apoptosis.</p>

Establishment and application of fluorescent quantitative PCR method for detecting human herpes virus type 6 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).</p><p><b>METHODS</b>According to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.</p><p><b>RESULTS</b>The standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.</p><p><b>CONCLUSION</b>The fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.</p>

Significance of detecting platelet associated antibody and platelet membrane glycoprotein for diagnosis of immune thrombocytopenia / 中国实验血液学杂志

The aim of this study was to explore application value of detecting platelet associated antibody and platelet membrane glycoprotein in the diagnosis and prognosis for immune thrombocytopenia. The platelet associated immunoglobulin (PAIg) and platelet membrane glycoprotein (CD41, CD61, GPIIb/IIIa) in 76 cases of immune thrombocytopenia and 30 healthy subjects were determined by FCM. The results showed that PAIg level in ITP patients included PAIgG (31.25 +/- 18.06)%, PAIgM (32.41 +/- 15.51)%, PAIgA (23.39 +/- 16.67)% which were remarkedly higher than in health control (10.48 +/- 5.05)%, (9.40 +/- 4.42)% and (7.23 +/- 3.61)% (P < 0.001). In patients with secondary immune thrombocytopenia (chronic aplastic anemia, SLE, Evans syndrome, liver cirrhosis hypersplenism, etc), PAIg level was higher than that in control group, while the platelet membrane glycoprotein in the blood of these patients was lower than that in control group. The level of PAIg decreased (P < 0.05) after treatment, but platelet membrane glycoprotein increased (P < 0.01). The result suggested that measurements for platelet membrane glycoprotein and platelet associated antibody by FCM were practical with high sensitivity, rapidity and simplicity used as a routine method in diagnosis and evaluation of the therapeutic effects in immune thrombocytopenia patients.