Expression of CD269 and CD317 in Multiple Myeloma and Its Clinical Significance / 现代检验医学杂志

Objective To study the expression of CD269 and CD317 antigens in bone marrow cells of patients with multiple myeloma (MM),analyze its correlation with the laboratory indexes reflecting the progression of MM and evaluate its value in clinical diagnosis and treatment.Methods 63 newly diagnosed MM patients were selected as the study group by a casecontrol study.The expression rate of CD269 and CD317 in bone marrow blood of 35 patients with iron deficiency anemia and other antigens in bone marrow blood of 63 patients with MM were detected by flow cytometry.The levels of serum hemoglo bin (Hb),serumβ2-MG(β2-MG) and lactatedehydrogenase (LDH) in patients with MM were dctectcd,and the levels of CD269 and CD317 were analyzed statistically.Results The positive rates of CD269 in the study group and control group were (86.6±2.35)％ vs (4.33±l.69)％,rcspectivcly (t =4.256,P＜0.05)).The positive rate of CD317 was (71.42+ 0.62)％ vs (8.32+ 3.89)％,the difference was statistically significant (t=3.102,P＜0.05).In other expression,the expression level of CD269 and CD317 in CD56 positive group was significantly higher than that of negative group (t=4.032,P＜0.05),while the expression of CD117 the level of positive group was significantly lower than that of the negative group (t 2.832,P＜0.05),CD19,CD20 expression was not statistically significant difference between the two groups (P＞ 0.05).The levels of CD269 and CD317 in patients with MM were positively correlated with the level of CD56 expression (r =0.392,P＜0.05),and negatively correlated with the level of CD117 expression (r=-0.210,P＜0.05).The levels of CD269 and CD317 in patients with MM were significantly lower than those in the negative group (t=3.012,P＜0.05) and the levels of serum LDH in the positive group were lower than those in the negative group (t=2.024,P＜0.05).There was a negative correlation between Hb content (r=-0.212,P＜0.05) and negatively correlated with serum β2-MG (r=-0.312,P＜0.05).Conclusion The high expression of CD269 and CD317 in bone marrow cells in MM patients is related to the increase of CD56 and decrease of CD117 in patients with MM.

Effect of Decitabine on Megakaryocyte Culture of Steroid-resistant ITP Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of decitabine and plasma of ITP patients on in vitro cultrue of megakaryocytes in bone marrow of steroid-resistant ITP patients.</p><p><b>METHODS</b>Bone marrow mononuclear cells were isolated from 20 steroid-resistant ITP patients, both methyl cellulose semisolid culture system (to observe and count the number of megakaryocytes colony-forming unit) and liquid culture system (to analysis the expression rate of CD41a(+) cells) were used for megakaryocyte cultrue. The experiments were divided into 4 groups according to the different components of the culture system, group A was control, group B was added with decitabine, group C with ITP plasma, group D with both decitabine and ITP plasma, and the rest of the culture components were the same in the 4 groups except the above-mentioned materials. Morphology of megakaryocytes was observed by inverted and light microscopy. The expression rate of CD41a⁺ cells in culture was analysed by flow cytometric.</p><p><b>RESULTS</b>Different concentration of decitabine showed different effect on megakaryocyte growth of steroid-resistant ITP patients and the optimal concentration to differentiate into megakaryocyte for bone marrow mononuclear cells is 3.0 µmol/L. Compared with group A, both megakaryocyte colony forming units (CFU) and expression rate of CD41a⁺ cells in group B were statistically significantly higher (P < 0.05). As compared with group A, the megakaryocyte colony-forming units in group C decreased with statistically significant difference, while compared with group C, the megakaryocyte colony-forming units in group D obviously increased with statistically significant difference.</p><p><b>CONCLUSIONS</b>Decitabine is able to induce bone marrow mononuclear cells of steroid-resistant ITP patients to differentiate into megakaryocyte and the optimal concentration is 3.0 µmol/L; ITP plasma is able to inhibit the megakaryocyte growth of steroid-resistant ITP patients.</p>

The frequency of JAK2 V617F mutation, expression level of phosphorylated JAK/STATs proteins and their clinical significance in myeloproliferative disorders patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the frequency of JAK2 V617F mutation in 145 myeloproliferative disorders (MPDs) patients, analyze the correlation between JAK2 V617F mutation and clinical features.</p><p><b>METHODS</b>The JAK2 V617F mutation was detected by direct DNA sequencing of PCR product and allele-specific PCR respectively. The expression of JAK2, phospho-JAK2 and phospho-STAT5 proteins was determined by Western blot. The clinical data of MPDs patients with or without JAK2 V617F mutation was collected and analyzed for evaluating the clinical significance of JAK2 V617F mutation.</p><p><b>RESULTS</b>1) The frequency of JAK2 V617F mutation for PV, IMF, ET was 92%, 58%, 50% respectively. Compared with conventional DNA sequencing (PV 84%, IMF 44%, ET 39%, respectively), allele-specific PCR exhibited a higher sensitivity in JAK2 V617F mutation detection. 2) The expression levels of phospho-JAK2 and phospho-STAT5 in peripheral blood mononuclear cells (PBMNCs) were upregulated significantly in JAK2 V617F-positive patients than in JAK2 V617F negative patients. 3) Compared with the patients with no JAK2 V617F mutation, the JAK2 V 617F-positive patients' features were as follows: older age of onset, higher mean leukocyte counts, lower platelet counts and smaller spleen volume. Frequency of thrombosis events in PT, ET, IMF was 17%, 32%, 16% respectively for JAK2 V617F positive group, and 0% (PV), 16% (ET), 5% (IMF) for JAK2 V617F negative group.</p><p><b>CONCLUSIONS</b>MPDs patients display higher frequency of JAK2 V617F mutation. JAK2 V617F mutation positive patients predispose to a thrombosis tendency.</p>

Analysis of angiogenesis related proteins and its implication in type-2 hereditary hemorrhagic telangiectasia / 中华血液学杂志

<p><b>OBJECTIVE</b>To detect the level of transforming growth factor-beta1 (TGF-beta1), TGF-beta2, vascular endothelial growth factor (VEGF) and platelet-derived growth factor receptor-alpha (PDGFRalpha) in plasma and peripheral blood leukocytes in a hereditary hemorrhagic telangiectasia type 2 (HHT-2) family, and explore the implication of angiogenesis related proteins in HHT-2 pathogenesis.</p><p><b>METHODS</b>The diagnosis of the HHT-2 patient was based on clinical features and further confirmed by determining a C1231T mutation of activin receptor-like kinase 1 (ALK1) gene. Five other new members in this family were evaluated with ALK1 gene screening and clinical manifestation. Plasma level of TGF-beta1, TGF-beta2 or VEGF was measured by ELISA, and the expression of PDGFRalpha,TGF-beta1, and VEGF in peripheral blood leukocytes by flow cytometry combined with direct or indirect immunofluorescence.</p><p><b>RESULTS</b>No C1231T mutation was detected in exon 8 of ALK1 gene in the 5 new members. Plasma TGF-beta1 and TGF-beta2 concentration in 3 affected HHT case was (16 954 +/- 3 709) ng/L and (11 548 +/- 2 611) ng/L, respectively, compared with that of normal control, the difference was not significant (P > 0.05). VEGF concentration in the 3 HHT patients, 6 unaffected family members and 6 normal controls was (179.2 +/- 22.0) microg/L, (149.8 +/- 22.7) microg/L and (132.9 +/- 21.0) microg/ L, respectively. Plasma VEGF level in HHT patients was significantly higher than that in normal subjects (P < 0.025). Peripheral leukocyte PDGFRalpha and VEGF in HHT patients and unaffected family members were markedly higher than that of normal control (P < 0.05 and P < 0.02), while TGF-beta1 distribution was similar in HHT patients and normal subjects.</p><p><b>CONCLUSION</b>Compared with normal controls there is no difference in plasma TGF-beta1 concentration on peripheral leukocytes of HHT patients. Plasma VEGF concentration or leukocytes VEGF expression in HHT is significantly higher than that of normal subjects. Leukocytes PDGFRalpha expression in HHT is significantly higher than that of normal control. These changes may be associated with a compensable mechanism in HHT.</p>