Karyotype complements the International Prognostic Scoring System for primary myelofibrosis

Objectives: The International Prognostic Scoring System (IPSS) for primary myelofibrosis (PMF) is based on five independent predictors of inferior survival: age >65 yr, hemoglobin <10 g/dL, leukocyte count >25 × 10⁹/L, circulating blasts ≥1%, and presence of constitutional symptoms. The presence of 0, 1, 2, and ≥3 adverse factors defines low, intermediate‐1, intermediate‐2, and high risk disease, respectively. We examined the additional prognostic relevance of karyotype. Methods: World Health Organization criteria were used for PMF diagnosis. Only patients with bone marrow cytogenetic studies at the time or within 1 yr of diagnosis and a minimum of 20 evaluable metaphases were considered. Cytogenetic findings were categorized as ‘normal’ vs. ‘abnormal’ or ‘favorable’ (normal or with sole abnormalities of 13q? or 20q?) vs. ‘unfavorable’ (all other abnormalities). Results: A total of 109 patients were studied (median age 63 yr). Numbers of patients in the above‐listed four IPSS risk groups were 26, 31, 28, and 24, respectively. Cytogenetic results were abnormal in 33% of the patients and unfavorable in 21%. At a median follow‐up of 35 months, 45 (41%) deaths were recorded. ‘Unfavorable’ (P = 0.008) but not ‘abnormal’ (P = 0.19) karyotype predicted shortened survival and its significance remained on multivariable analysis that included the IPSS or other prognostic tools as covariates. JAK2V617F, detected in 63 (58%) patients, was inconsequential to survival. Conclusions: In PMF, specific cytogenetic abnormalities and not the mere presence of an abnormal karyotype provide important prognostic information that is not accounted for by the IPSS or other established risk factors.     

Postpolycythaemic myelofibrosis: frequency and risk factors for this complication in 116 patients

Postpolycythaemic myelofibrosis (PPMF) is a known complication of polycythaemia vera (PV) but information regarding its incidence and predisposing factors is not well defined. In 116 subjects consecutively diagnosed with PV in a single institution (median age 62 years, range: 20–88), the probability of PPMF was analysed by the Kaplan–Meier method, followed by the log-rank test. With a mean follow-up of 8 years (95% confidence interval: 6·6–9), 17 patients had evolved into PPMF (15%). The probability of evolution to PPMF was 16% at 10 years and 34% at 15 years. Age, gender, spleen size, leucocytosis, thrombocytosis or cytoreductive treatment were not associated with an increased risk of PPMF. The actuarial probability of PPMF at 15 years was higher in those patients presenting at diagnosis with endogenous megakaryocytic colony formation (59% when present versus 10% when absent, P  = 0·03), an elevated serum lactate dehydrogenase (LDH) level (69% vs. 23% in patients with normal LDH, P  = 0·04), and in those who were heterozygous for the JAK2 V617F mutation (55% vs. 17% in heterozygotes, P  = 0·04). In conclusion, PPMF is a frequent complication in PV patients at 15 years with the risk being higher in patients with increased LDH, endogenous megakaryocytic colony formation or a high JAK2 V617F allele burden.     

辐照与去白细胞过滤单采血小板对血小板诸参数与部分细胞因子含量影响

目的：了解应用去白细胞过滤器制备去白细胞单采血小板以及经γ射线辐照单采血小板血小板参数与血浆部分细胞因子含量变化情况。方法：应用SysmexF-800血液分析仪检测血小板参数与应用ELISA法检测血浆IL-2、IL-6、INF-γ与TNF-α细胞因子。结果：去白细胞单采血小板与经γ射线辐照单采血小板悬液血小板包括：血小板数、血小板平均体积、血小板平均宽度、血小板压积以及血浆IL-2、IL-6、INF-γ与TNF-α含量与对照组相互比较差异均无统计学意义（P〉0．05）。结论：单采血小板经去白细胞或经γ射线辐照后,不会影响血小板参数与血浆部分细胞因子含量。     

Anaemia characterises patients with myelofibrosis harbouring Mpl mutation

The clinical and haematological phenotype of patients with myelofibrosis harbouring MPL(W515L/K) mutation has not been thoroughly investigated. Of 217 myelofibrosis subjects, 18 (8.2%) had an MPL mutation, four of which (22%) co-existed with JAK2(V617F) mutation. When compared with MPL wild-type patients, irrespective of JAK2(V617F) status, those with MPL(W515L/K), were more frequently female, were older (61 years vs. 57 years; P = 0.02), presented with more severe anaemia (haemoglobin, 101 g/l vs. 121 g/l; P = 0.002) and were more likely to require regular transfusional support (P = 0.012). These data indicate that MPL mutation in myelofibrosis characterises patients with more severe anaemic phenotype.     

Red blood cell transfusion-dependency implies a poor survival in primary myelofibrosis irrespective of IPSS and DIPSS

Risk stratification in primary myelofibrosis is currently based on two international prognostic scoring systems, neither of which takes into consideration red blood cell transfusion-dependency. In 288 consecutive patients with primary myelofibrosis, red blood cell transfusion-dependency at diagnosis affects survival independently of the International Prognostic Scoring System (P < 0.001). To evaluate the dynamic impact on survival of red blood cell transfusion-dependency, we performed a Cox's regression analysis with transfusion status as time-dependent covariate in 220 regularly followed patients with primary myelofibrosis. Patients who begin red blood cell transfusions anytime (n = 80, 36%) have a significantly worse survival compared to those who continue follow up without transfusions (HR: 7.8, 95%CI: 5.1-11.9; P < 0.001). Adjusting for Dynamic International Prognostic Scoring System in a multivariate analysis, red blood cell transfusion-dependency retained an independent prognostic impact on survival. This study suggests that red blood cell transfusion-dependency should be considered to improve risk stratification of primary myelofibrosis during follow up.     

Toxic epidermal necrolysis in a patient with primary myelofibrosis receiving thalidomide therapy

Primary myelofibrosis (PMF) is a chronic myeloproliferative neoplasm characterized by progressive anemia, massive splenomegaly, leukoerythroblastosis, extramedullary hematopoiesis and in about 50% of cases the presence of JAK2V617F mutation. Curative therapy in PMF is currently possible only with allogeneic haematopoietic stem cell transplantation which is, unfortunately, associated with relatively high risks of mortality and morbidity which undermine its broad applications. Non-transplant treatment modalities are used for palliative purposes. Recently, anti-angiogenic drugs such as thalidomide have been used to treat these patients on the basis of the prominent bone marrow angiogenesis. Here, we report the case of a patient suffering from JAK2V617F-positive PMF with marked bone marrow neo-angiogenesis. The patient was treated with thalidomide but after 20 days developed life-threatening toxic epidermal necrolysis (TEN). To the best of our knowledge this is the first case of TEN in a patient with PMF under thalidomide therapy.     

Intestinal obstruction caused by extramedullary hematopoiesis and ascites in primary myelofibrosis

Primary myelofibrosis (PMF) is a clonal hematopoietic stem cell disorder. It is characterized by bone marrow fibrosis, extramedullary hematopoiesis with hepatosplenomegaly and leukoerythroblastosis in the peripheral blood. The main clinical manifestations of PMF are anemia, bleeding, hepatosplenomegaly, fatigue, and fever. Here we report a rare case of PMF with anemia, small bowel obstruction and ascites due to extramedullary hematopoiesis and portal hypertension. The diagnosis was difficult to establish before surgery and the differential diagnosis is discussed.     

血小板活化在哮喘发病中的意义

目的　探讨血小板活化在哮喘发病中的作用。方法　利用酶联免疫法及流式细胞仪分别测定 6 2例哮喘患者和 18名正常对照者血浆 11 去氢血栓烷B2 (11 DH TXB2 )、血栓烷 (TXB2 )浓度和血小板膜CD 6 2P的表达情况 ,并利用荧光酶联免疫法测定部分血清嗜酸细胞阳离子蛋白 (ECP)水平。结果　症状性哮喘组血浆 11 DH TXB2 浓度为 (33 2± 2 7)ng/L ,血小板CD 6 2P阳性百分比为 (2 3 8±3 0 ) % ,平均荧光强度为 2 75± 0 2 9;与健康对照组和缓解组比较差异有显著性 (P分别 <0 0 1、<0 0 5 )。缓解期哮喘与健康对照组比较仅血浆 11 DH TXB2 浓度差异有显著性 (P <0 0 5 )。哮喘发作时血小板活化水平与血清ECP及一秒钟用力呼气容积占预计值百分比 (FEV1占预计值百分比 )、最大呼气流量 (PEF)相关。结论　哮喘发作时存在血小板的异常活化 ,其活化程度与哮喘气道炎症及病情有一定相关性 ,血小板可能通过影响嗜酸细胞功能参与哮喘的发病     

A compendium of cytogenetic abnormalities in myelofibrosis: molecular and phenotypic correlates in 826 patients

Among 826 patients with primary myelofibrosis (PMF) and analysable metaphases on cytogenetic studies, 352 (42·6%) had abnormal karyotype, of which 240 (68·2%) were sole aberrations and 48 (13·6%) were complex; the most frequent abnormalities were 20q? (23·3%), 13q? (18·2%), +8 (11·1%), +9 (9·9%), chromosome 1q+ (9·7%) and ?7/7q? (7·1%). Phenotypic correlates included: abnormal karyotype with anaemia (P = 0·02), leucopenia (P < 0·01) and thrombocytopenia (P < 0·01); complex karyotype with younger age (P = 0·04) and thrombocytopenia (P < 0·01); leucopenia with 20q?, +8 and ?7/7q? and thrombocytopenia with 20q? and ?7/7q?. Cytopenias were less likely to occur with 13q?. 476 patients were annotated for JAK2/CALR/MPL mutations; abnormal karyotype frequencies were 43% in JAK2, 42% CALR, 33% MPL mutated and 34% triple‐negative cases (P = 0·3). A proportion of patients were also screened for ASXL1, EZH2, IDH1, IDH2, SRSF2, U2AF1 and SF3B1 mutations; in all instances, mutational frequencies were higher in patients with normal karyotype, reaching significance for ASXL1 (P = 0·02) and U2AF1 (P = 0·01). 13q? was associated with mutant CALR (P = 0·03), +9 with mutant JAK2 (P = 0·02) and 20q? with mutant SRSF2 (P = 0·02). The current PMF study provides detailed cytogenetic information and correlations with mutations and clinical phenotype.     

The significance of platelet activation in ankylosing spondylitis

The objective of this study was to explore the significance of platelet activation in patients with ankylosing spondylitis (AS). Thirty-five AS patients and 15 normal controls were selected from November 2005 to October 2006. The number of CD62P- and CD63-positive cells were detected by flow cytometry. At the same time, the erythrocyte sedimentation rate (ESR), platelet count (PLT) and C-reactive protein (CRP) were determined in both groups. The percentage of CD62P-positive cell in AS patients (13.60 ± 7.64%) was significantly higher than that in control group (2.78 ± 1.04%; P < 0.01). The percentage of CD63-positive cell in AS patients (6.92 ± 4.16%) was significantly higher than that in control group (4.13 ± 1.85%; P < 0.05). The levels of CRP (20.18 ± 23.17 mg/l), PLT (259.54 ± 102.59 × 10⁹/l) and ESR (36.86 ± 31.23 mm/h) in AS patients were higher than those in normal controls, respectively (3.21 ± 2.18 mg/l, P < 0.01; 197.00 ± 55.70 × 10⁹/l, P < 0.01; 12.25 ± 5.05 mm/h, P < 0.05). Platelet activation may be a sign of AS exacerbation.     

Increased blood coagulation and platelet activation in patients with infective endocarditis and embolic events

BACKGROUND: Inflammation-induced procoagulant changes and alterations in platelet activity appear to play an important role in thromboembolic complications of infective endocarditis (IE). HYPOTHESIS: The aim of this study was to investigate systemic coagulation activity, fibrinolytic capacity, and platelet activation in patients with IE with and without embolic events by measuring the plasma levels of prothrombin fragment 1+2 (PF1+2), thrombin-antithrombin III complex (TAT), plasminogen activator inhibitor-1 (PAI-1), beta-thromboglobulin (beta-TG), and platelet factor 4 (PF4), respectively. METHODS: The study included 76 consecutive patients (female = 55, male = 21, mean age 26 years, range 8-64 years) with definite IE according to the Duke criteria; of these, 13 (17.1%) had embolic events. RESULTS: Plasma concentrations of PF1+2 (3.2 +/- 1.3 vs. 1.7 +/- 0.7 and 1.4 +/- 0.7 nmol/l, p < 0.001, respectively) and TAT (7.3 +/- 1.5 vs. 2.9 +/- 1.2 and 2.2 +/- 1.1 ng/ml, p < 0.001, respectively) were elevated in patients with embolic events compared with patients without embolic events and control subjects. Similarly, patients with embolic events had increased plasma levels of beta-TG (63.3 +/- 10.9 vs. 33.1 +/- 11.6 and 19.1 +/- 10.6 ng/ml, p < 0.001, respectively) and PF4 (106.0 +/- 28.7 vs. 50.3 +/- 16.7 and 43.0 +/- 15.8 ng/ml, p < 0.001, respectively) compared with those without embolic events and the control group. Embolic patients also had higher PAI-1 levels than nonembolic patients and healthy subjects (14.4 +/- 6.4 vs. 8.6 +/- 5.9 and 5.4 +/- 4.3 ng/ml, p = 0.002, respectively). CONCLUSION: Patients with IE and with subsequent thromboembolism have increased systemic coagulation activation, enhanced platelet activity/damage, and impaired fibrinolysis. The resulting imbalance produces a sustained hypercoagulable state, which contributes to the increased risk of thromboembolic events in this particular group.     

Plasma soluble interleukin-2 receptor in patients with primary myelofibrosis

Summary Using an enzyme-linked immunosorbant assay (ELISA) test, the level of soluble Tac peptide, one chain of the human interleukin-2 receptor, was measured in the plasma of 26 patients with primary myelofibrosis (MF), seven patients with polycythaemia vera and 11 normal controls. The plasma soluble interleukin-2 receptor (sIL-2R) was found to be significantly elevated in patients with primary MF compared to polycythaemia vera or controls ( P <0·01), while the plasma sIL-2R of patients with polycythaemia vera also was found to be significantly elevated compared to controls ( P <0·01). The significantly elevated value of sIL-2R seen in primary MF may be secondary to T cell activation resulting from autoimmune phenomena, and myeloblast activation with release of sIL-2R may also be a contributing factor. In primary MF, plasma sIL-2R levels were also found to be correlated to survival, circulating blast cell counts, and thrombocytopenia, but not to white blood cell counts, LDH levels, degree of marrow fibrosis, or degree of splenomegaly. Patients with primary MF with higher titre of plasma sIL-2R had a shorter survival. Further studies involving more patients and longer follow-up may substantiate that plasma sIL-2R is an important prognostic indicator in primary MF.     

活动期溃疡性结肠炎患者体内血小板激活状态的评价

目的 探讨血小板激活和活动期溃疡性结肠炎的关系。方法 对32例活动期溃疡性结肠炎、11例缓解期溃疡性结肠炎、30例肠易激综合征（IBS）患者和正常对照组27例用SH－93智能血液凝聚仪检测血小板聚集率，用酶联免疫法检测P－选择素和血栓素B2（TXB2）。同时评价43例溃疡性结肠炎内镜下表现和结肠黏膜活检情况。结果 32例活动期溃疡性结肠炎患者平均1min血小板聚集率和最大血小板聚集率均明显高于IBS组和正常对照组（P＜0．01）；32例活动期溃疡性结肠炎P－选择素和TXB2明显高于IBS组和正常对照组，差异有显著性（P＜0．01或P＜0．05）。缓解期溃疡性结肠炎的P－选择素也高于正常对照组。结论 活动期溃疡性结肠炎患者体内存在血小板激活，血小板可能直接参与结肠黏膜的急性炎症反应。其中P－选择素是溃疡性结肠炎的特异性指标，而血小板聚集率和TXB2与疾病的活动度有关。抗血小板药物可能对溃疡性结肠炎有一定的治疗效果。     

Increased granulocytic, erythrocytic, and megakaryocytic progenitors in myelofibrosis with myeloid metaplasia

Nucleated cells obtained from blood and/or bone marrow of patients with myelofibrosis with myeloid metaplasia (MMM) were cultured in diffusion chambers (DC) implanted into the peritoneal cavities of irradiated mice. A total of five blood studies and two bone marrow studies were performed using cells obtained from five patients. The DC were harvested at intervals from the host mice and the total and differential cellularity of DC contents were evaluated. The results obtained from MMM cultures were compared with those from similar cultures of blood cells and marrow cells of four and six normal individuals respectively. The proliferation and maturation of the granulocytic, erythrocytic, and megakaryocytic lines in MMM cultures occurred in an orderly fashion as they occur in vivo. The patterns of proliferation and maturation of the three cell lines in cultures after day 7 suggest that they primarily originate from progenitor cells. The numbers of granulocytes in the multiplicative pool, recognizable red cell precursors, and megakaryocytes recovered were significantly greater from the MMM cultures than those from the normal blood or marrow cultures. These results suggest that the blood and marrow cells of MMM patients have increased numbers of progenitors for granulocytes, erythrocytes, and megakaryocytes.     

The significance of platelet activation in rheumatoid arthritis

We evaluated the significance of platelet activation in patients with rheumatoid arthritis (RA). The expression of CD62P and CD63 by platelets was determined using flow cytometry in 18 active RA patients, 10 remission RA and 15 normal controls. Meanwhile, the erythrocyte sedimentation rate (ESR) and C-reactive protein was also determined in all groups. The expression of CD62P in active RA patients (11.88 ± 2.47%) was significantly higher than that in remission RA group (2.85 ± 1.60%; P < 0.01) and control group (2.78 ± 1.04%; P < 0.01). The expression of CD63 in active RA patients (9.90 ± 3.02%) was significantly higher than that in remission RA group (4.11 ± 2.00%; P < 0.01) and control group (4.13 ± 1.85%; P < 0.01). The level of CRP (54.33 ± 23.35 mg/l) and ESR (86.06 ± 33.67 mm/h) in active RA patients was higher than that in remission RA group (2.55 ± 1.01 mg/l, 14.70 ± 4.57 mm/h; P < 0.01 for both) and normal control group (3.21 ± 2.18 mg/l, 12.25 ± 5.05 mm/h; P < 0.01 for both). There was a positive correlation between CD62P and ESR (r = 0.5224, P < 0.01) and also a positive correlation between CD62P and CRP (r = 0.7048, P < 0.01) as well as between CD63 and ESR (r = 0.4476, P < 0.05) but no correlation between CD63 and CRP. Platelet activation may be a sign of RA exacerbation.