The morphology features of bone marrow in the prefibrotic-early primary myelofibrosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To compare the morphologic features of bone marrow (BM) between the prefibrotic-early primary myelofibrosis (PMF) and essential thrombocythaemia (ET).</p><p><b>METHOD</b>Seven cases of prefibrotic-early PMF were selected and analyzed. Based on the diagnostic standard of prefibrotic-early PMF by WHO, BM aspirate smears, trephine biopsy sections and imprints of 156 uncertain ET cases conducted simultaneously were recruited into this study, the BM morphologic features between the prefibrotic-early PMF and ET groups were analyzed. The morphological difference in 22 cases of prefibrotic-early PMF and 27 ET were compared between the JAK2V617F mutation positive and negative groups.</p><p><b>RESULTS</b>Of the 156 uncertain ET cases, it was reclassified 61 prefibrotic-early PMF (34 MF-0, 27 MF-1), 12 PMF and 83 ET. The platelet count and LDH level in MF-1 group were obviously higher than that of ET group (P < 0.05). The blast percentage of BM smear in MF-1 group was also higher than that of ET group (P < 0.05). As to BM section, cases with increased nucleated cells (granulocyte), compact megakaryocytic cluster, megakaryocyte near bone trabecula, cloud-like megakaryocyte, small bare nucleus of megakaryocyte and large ball-like megakaryocyte in MF-0 and MF-1 group were significantly higher than that of ET group (all P < 0.05), cases with megakaryocytic cluster of various size in MF-1 group were significantly higher than that of MF-0 and ET groups (P < 0.05). The JAK2V617F mutation rate in prefibrotic-early PMF and ET groups were 54.5% and 48.1%, respectively. Hb level in JAK2V617F mutation positive group was obviously higher than the negative group (P < 0.05), no special change with megakaryocytic morphology was found between the positive and negative groups.</p><p><b>CONCLUSION</b>Morphology of BM section, especially megakaryocytic morphologic characteristics are the main basis in distinguishing prefibrotic-early PMF from ET. The importance of morphologic index were megakaryocytic cluster with various size, cloud-like megakaryocyte, large ball-like megakaryocyte, increased nucleated cells (granulocyte), small bare nucleus, megakaryocyte near bone trabecula and compact megakaryocytic cluster in order. JAK2V617F mutation provides no specific effect on the megakaryocytic morphology.</p>

Morphological observation on bone marrow megakaryocytes in patients with bacterial and fungal infection / 中华临床感染病杂志

Objective To investigate the morphological changes of bone marrow megakaryocytes in patients with bacterial and fungal infection.Methods Totally 76 patients with microorganism infection from the Second Affiliated Hospital,Zhejiang University School of Medicine from January 2008 to August 2009 were enrolled,including 56 bacteria infected patients and 20 fungal infected patients.All patients received bone marrow examinations,and were positive in microorganism culture.Thirty subjects without infection,hematological disease and other severe diseases were randomly selected as controls.The number and function of megakaryocytes were examined retrospectively, and the size, nuclear lobulation, and vacuolar degeneration of megakaryocytes were quantitative analyzed and compared among the groups.Results The size,nuclear lobulation,vacuolar degeneration,and Yat nuclear of megakaryocytes in bacterial infected group were 2.20 ±0.21,2.11 ±0.23,0.51 ±0.11 and 0.74 ±0.11 respectively,those in fungal infected group were 2.21 ±0.16,2.10 ±0.19,0.52 ±0.10 and 0.79 ±0.10 respectively;while those in control group were 1.40 ±0.10,1.36 ±0.12,0.28 ±0.06 and 0.54 ±0.09 respectively.The differences between bacterial infected group and control were of statistical significance(t values were 14.52,12.19,9.33 and 6.61 respectively,P ＜ 0.05),and the differences between fungal infected group and control were of statistical significance(t values were 16.27,12.34,7.85 and 6.49 respectively,P ＜ 0.05).The size,nuclear lobulation,and vacuoles of megakaryocytes in gram-negative(G-)bacteria group were 2.29 ±0.20,2.22 ±0.26 and 0.57 ±0.10,while those in the gram-positive(G+)bacteria group were 2.13 ±0.20,2.04 ±0.18 and 0.46 ±0.09,and the differences were also significant(t values were 2.07,3.03and 3.56 respectively,P ＜ 0.05).The production of platelet by megakaryocytes in bacterial infected group,in fungal infected and the control were 31.4 ±7.6,32.4 ±6.4 and 41.3 ±5.5,and the differences between bacterial infected group and control,fungal infected group and control were significant(t values were 4.78and 3.98 respectively,P ＜ 0.05).The production of platelet in G-bacteria group was 28.0 ± 6.7,while that in G + bacteria group was 34.4 ± 7.2,and the difference was also of statistical significance(t = 2.41,P ＜0.05). Conclusion Bacterial infected patients have increased megakaryocytes cell body,nuclear lobulation,obvious vacuolar degeneration,Yat nuclear and decreased platelet production function,which are more significant in G- bacteria infected group.

Improvement of Phi bodies stain and its clinical significance / 中国实验血液学杂志

The aim of this study was to improve the dyeing method of hydroperoxidase (HPO), to analyze the morphologic features of Phi bodies and to evaluate the clinical application of this method. 128 bone marrow or peripheral blood smears from patients with myeloid and lymphoid malignancies were stained by improved HPO staining. The Phi bodies were observed with detection rate of Phi bodies in different leukemias. 69 acute myeloid leukemia (AML) specimens were chosen randomly, the positive rate and the number of Phi bodies between the improved HPO and POX stain based on the same substrate of 3, 3'diaminobenzidine were compared. The results showed that the shape of bundle-like Phi bodies was variable, long or short. while the nubbly Phi bodies often presented oval and smooth. Club-like Phi bodies were found in M(3). The detection rates of bundle-like Phi bodies in AML M(1)-M(5) were 42.9% (6/14), 83.3% (15/18), 92.0% (23/25), 52.3% (11/21), 33.3% (5/15) respectively, and those of nubbly Phi bodies were 28.6% (4/14), 66.7% (12/18), 11.1% (3/25), 33.3% (7/21), 20.0% (3/15) respectively. The detection rate of bundle-like Phi bodies in M(3) was significantly higher than that in (M(1) + M(2)) or (M(4) + M(5)) groups. The detection rate of nubbly Phi bodies in (M(1) + M(2)) group was higher than that in M(3) group. In conclusion, after improvement of staining method, the HPO stain becomes simple, the detection rate of Phi bodies is higher than that by the previous method, the positive granules are more obvious, and the results become stable. This improved method plays an important role in differentiating AML from ALL, subtyping AML, and evaluating the therapeutic results.

Clinical study of plasma thrombomodulin detection / 中国实验血液学杂志

The purpose of this study was to investigate the clinical value of plasma thrombomodulin (PTM) in different diseases or in different severity or complications of diseases, PTM in 979 patients and 60 healthy controls was determined by ELISA method. The results showed that the PTM level in the control group was 20.40 +/- 7.72 microg/L, there was no difference in sex and ages. In chronic primary glomerular disease, the PTM level in chronic renal failure (CRF) group was higher than that in non-CRF group (P < 0.01). PTM level > 70 microg/L was defined as its positive criterion. The sensitivity, specificity and positive predictive value in PTM were 85.7%, 82.4% and 77.8% respectively. The PTM level in septemia group was higher than that in non-septemia group (P < 0.01), the sensitivity, specificity and positive predictive value were 86.6%, 89.5% and 76.5% respectively (> 50 microg/L as its positive criterion). With respect of multiple trauma, the PTM level in multiple organ failare (MOF) group was higher than that in non-MOF group (P < 0.01), while the sensitivity, specificity and positive predictive value were 77.8%, 77.3% and 73.7% respectively (> 40 microg/L as its positive criterion). For systemic lupus erythematosus (SLE), the PTM level in the patients with albuminuria was higher than that in the patients without albuminuria (P < 0.01), and the sensitivity, specificity and positive predictive value were 77.8%, 92.3% and 93.3% respectively (> 35.54 microg/L as its positive criterion). For diabetes, the PTM level in complication group was higher than that in group without complications, the sensitivity, specificity and positive predictive value were 53.4%, 97.1% and 98.6% respectively (> 35.54 microg/L as its positive criterion). The PTM level in microangiopathy group was higher than that in macroangiopathy group (P < 0.01). The sensitivity, specificity and positive predictive value were 71.2%, 97.1% and 97.9% respectively. Acute leukemia (AL) and multiple myeloma (MM) had higher PTM level and PTM level was extremely high when renal failure developed (P < 0.01). As compared the acute stage with the restoration stage in stroke, pre-chemotherapeutics with post-chemotherapeutics in AL and MM, and pre-operation with post-operation in cancer, the PTM level was connected with clinical development. The PTM level in the patients with microangiopathy was higher than that in the patients with macroangiopathy (P < 0.01). The defined PTM level was higher than its normal upper limit as PTM positive criterion in microangiopathy diseases, the sensitivity, specificity and positive predictive value were 77.7%, 71.2% and 75.6% respectively. It is concluded that PTM level is a good criterion in evaluating the microangiopathy, and PTM is also a valuable indicator in prediction or assessment of the severity of diseases, or evaluation of therapeutic effectiveness.

Cerulenin changes apoptosis related genes expression in multiple myeloma cell line U266 / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the expression changes of apoptosis related genes induced by cerulenin in multiple myeloma cell line U266 and explore its molecular mechanism.</p><p><b>METHODS</b>The expression changes of 96 apoptosis related genes were analyzed by superArray cDNA in U266 cells treated with cerulenin (20 microg/ml) for 12 h. Semi-quantitative RT-PCR was used to confirm the representative expression changes genes, Rip2, caspase 9 and TRAF2.</p><p><b>RESULTS</b>After treated with cerulenin for 12 h, 44 apoptosis related genes expression in the U266 cells were changed, among which 41 were over 2 fold increase and 3 over 2 fold decrease. The expression of caspase 9 was increased markedly, indicating that mitochondria pathway played a key role in cerulenin inducing apoptosis and TRAF2 expression change suggested that nuclear factor (NF) participates in cerulenin inducing apoptosis.</p><p><b>CONCLUSION</b>The death acceptor signaling pathway and the death acceptor non-dependence signaling pathway co-regulate cerulenin inducing apoptosis in U266 cells. Mitochondria pathway played the key role and nuclear factor (NF) participates in the apoptosis process.</p>

Novel distribution pattern of fibrinolytic components in rabbit tissues extract: a preliminary study / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>The purpose of this work was to investigate the distribution pattern of fibrinolytic factors and their inhibitors in rabbit tissues.</p><p><b>METHODS</b>The components of the fibrinolytic system in extracts from a variety of rabbit tissues, including tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and alpha(2) plasmin inhibitor (alpha(2)PI), were determined by colorimetric assay.</p><p><b>RESULTS</b>The tissue extracts in renal, small intestine, lung, brain and spleen demonstrated strong fibrinolytic function, in which high activity of tPA, Plg and Pl was manifested; whereas in skeletal muscle, tongue and stomach, higher activity of PAI-1 and alpha(2)PI showed obviously. Also excellent linear correlations were found between levels of tPA and PAI-1, Pl and alpha(2)PI, Plg and Pl. In related tissues, renal cortex and renal marrow showed distinctly higher activity of tPA and lower activity of PAI-1, with the levels of Plg and Pl in renal cortex being higher than those in renal marrow, where the alpha(2)PI level was higher than that in renal cortex. Similarly, the levels of tPA, Plg and Pl in small intestine were higher than those in large intestine, but with respect to PAI-1 and alpha(2)PI, the matter was reverse. In addition, the fibrinolytic activity in muscle tissue was lower, however, the levels of tPA, Plg, and Pl in cardiac muscle were obviously higher than those in skeletal muscles, and the levels of PAI-1 and alpha(2)PI were significantly lower than those in skeletal muscle.</p><p><b>CONCLUSION</b>Our data demonstrate that a remarkable difference of the fibrinolytic patterns exists in rabbit tissues, which has probable profound significance in understanding the relationship between the function of haemostasis or thrombosis and the physiologic function in tissues.</p>