Inhibitory effect of 4-chlorobenzoyl berbamine on imatinib-resistant K562 cells in vitro and in vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.</p><p><b>METHODS</b>The IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.</p><p><b>RESULTS</b>The IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.</p><p><b>CONCLUSION</b>BBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.</p>

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.

Following-up of nosocomial lower respiratory infection in patients with hematological malignancy after chemotherapy / 中华预防医学杂志

<p><b>OBJECTIVE</b>To observe and investigate the risk factors and pathogen diversification of nosocomial lower respiratory infections in patients with hematological malignancy after chemotherapy.</p><p><b>METHODS</b>Respiratory tract microbial population of fifty patients with different kinds of hematological malignancy and para-prepared to chemotherapy was quantitatively analyzed before and after chemotherapy at an arranged time from April, 2004 to December, 2005. Susceptibility test was determined for bacterium of nosocomial infection, and the homology of the same species of the bacteria was analyzed by a pulsed field gel electrophoresis (PFGE).</p><p><b>RESULTS</b>Incidence rate of lower respiratory infections in patients with the hematological malignant after chemotherapy was 16%. The major nosocomial infectious pathogens were Acinetobacter spp; Escherichia coil and Fungus. Among them, Acinetobacter spp, were highly resistant to cephalosporins, quinolones, aminoglycosides, carbapenems and antibiotic with enzyme inhibitor, respectively but susceptible to Cefoperazone/Sulbactam belonging to antibiotic with enzyme inhibitor. And it was shown that there were two clones by the pulsed field gel electrophoresis (PFGE).</p><p><b>CONCLUSION</b>Following-up of nosocomial lower respiratory infection in patients with hematological malignancy after chemotherapy might offer theoretical evidence for the rational use of antibiotics and the control of nosocomial infections.</p>

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.

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>

Application of bone marrow biopsy imprint in evaluating cellularity / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the value of bone marrow biopsy imprint in evaluating cellularity.</p><p><b>METHODS</b>The bone marrow tissues were obtained by trephine biopsy from 272 patients, and then put on the slides to make the imprints. The imprints was stained by Wright-Giemsa method, and the bone marrow smears and imprints were examined simultaneously according to the bone marrow cellularity criteria.</p><p><b>RESULT</b>In bone marrow cellularity, four grades (distinct decrease, extreme decrease, distinct increase, and extreme increase) were significantly higher in bone marrow imprints than those in bone marrow smears (P <0.05), but there was no significantly differences between bone marrow imprints and sections (P >0.05). Using bone marrow sections as standard, in cellularily decreasing samples, the consistent rate of bone marrow imprints and smears were both high (84.4% and 97.9%), in the group of the normal and increased cellularity, the consistent rate of the bone marrow imprints (84.4% and 97.7%) was significantly higher than that in smears (60% and 64%, P<0.01). The sensitivity, specificity, Youden index, positive predictive value and positive likelihood rate of bone marrow imprints were all higher than those of the smears. Using the bone marrow sections as gold standard, in 124 cases with decreased cellularity in smears, the positive diagnosis rate for aplastic anemia and dyshaematopoiesis based on bone marrow imprints was 37.1% with a false positive rate of 7.3% which was lower than that of the bone marrow smears (false positive rate of 29.8%, P<0.01).</p><p><b>CONCLUSION</b>To evaluate bone marrow cellularity, bone marrow imprint is better than bone marrow smear. The combination of the two examinations can make the diagnosis more convenient and quicker.</p>

Morphological study on the megakaryocytes with nuclear extrusion and nucleocytoplasmic separation in four cases / 中国实验血液学杂志

To investigate the morphological changes of megakaryocytes with nuclear extrusion and nucleocytoplasmic separation, the morphological characteristics of megakaryocytes in peripheral blood films, bone marrow smears, and bone marrow biopsies from 4 newly diagnosed patients with primary myelofibrosis (PMF), myelodysplastic syndrome (MDS), myeloblastic leukemia with maturation (M(2)) and erythroleukemia (M(6)) were studied by using light microscope. The results showed that many kinds of dysmegakaryocytes were observed in bone marrow smears of 4 cases, while in case A (PMF) and case D (M(6)) micromegakaryocytes were ripped apart; in case B (MDS) and case C (M(2)) megakaryocytes were accompanied by nuclear extrusion or nucleocytoplasmic separation, and their bodies were large or giant, the part of nucleus separated from their body and little cytoplasm remained as micromegakaryocytes. The nucleocytoplasmic separation could be displayed by immunocytochemistry stain. It is concluded that the phenomenon of nuclear extrusion and nucleocytoplasmic separation in megakaryocytes suggested the process that dispersed multinuclear releasing towards surround or even totally left the cell body during the megakaryocyte maturation. It also showed that the micromegakaryocytes may be the result of nucleocytoplasmic separation or splittings from multi-separated nucleus.

Detection of thrombomodulin in both plasma and tissue extracts of cancer patients and its clinical significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the changes of thrombomodulin(TM) in both plasma and tissue extracts of cancer patients for evaluating its clinical significance.</p><p><b>METHODS</b>Plasma TM levels were measured by enzyme-linked immunosorbent assay(ELISA) in plasma of 188 cancer patients and in 24 cancer tissue extracts including their adjacent normal tissue.</p><p><b>RESULTS</b>The plasma TM levels both in cancer patients and in metastasis patients were significantly higher than that in controls [(33.47+/-14.25) microg/L/ (41.68 +/-16.96) micro/L, compared with (20.40+/-7.22) microg/L, P<0.01]. The plasma TM levels in cancer patients after operation decreased obviously [(18.45+/-9.96) microg/L, compared with (28.29+/-11.74) microg/L,P<0.01]. Whereas, the plasma TM levels in patients with recurrence and metastasis after operation increased obviously [(34.50+/-12.57 micro/L]. The plasma TM levels in metastasis of lung cancers, gastric cancers and pancreatic cancers were significantly higher than that in non-metastasis (P<0.05 approximate, equals 0.01) respectively, but no significant differences were found between controls and non-metastasis cancers including gastric cancers, pancreatic cancers, nasopharyngeal cancers, large intestine cancers and laryngeal cancers (P>0.05). The TM levels in cancer tissue extracts were significantly lower than that in their adjacent normal tissue extracts [(647.71+/-317.51)) microg/L,compared with (1455.63+/-772.22) microg/L,P<0.01]. On the contrary, the plasma TM levels in these cancers were higher than that in controls.</p><p><b>CONCLUSION</b>The rise of plasma TM levels in cancer patients is associated with metastasis and diffusion of cancers. The TM levels can be used as an sensitive index for judging progression and metastasis of cancers.</p>

Detection of soluble Apo-1/Fas in plasma, pleural and ascites fluid of malignant tumor patients and its clinical significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the changes of soluble Apo-1/Fas levels in plasma, pleural and ascites fluid of malignant tumor patients and to evaluate their clinical significance.</p><p><b>METHODS</b>The soluble Apo-1/Fas levels were measured by enzyme-linked immunosorbent assay (ELISA) in the plasma of 157 malignant tumor patients and 25 normal controls as well as in the pleural and ascite fluids of 129 patients with various diseases.</p><p><b>RESULT</b>The plasma soluble Apo-1/Fas levels in acute and chronic leukemia and multiple myeloma were significantly higher than those in normal controls (P <0.05). The plasma soluble Apo-1/Fas levels in chronic myeloid leukemia and chronic lymphocytic leukemia were significantly higher than those in acute myeloid leukemia and acute lymphocytic leukemia, respectively (P <0.05). After chemotherapy, the plasma soluble Apo-1/Fas levels in complete remission group were distinctly decreased(P <0.05),whereas the levels in no remission and recurrence groups remained high. Compared with normal controls, the plasma soluble Apo-1/Fas levels in solid tumors were significantly increased (P <0.01), and the levels in metastasis cancers were significantly higher than those in non-metastasis cancer (P <0.0 1). Simultaneously the levels in remission cancer patients after operation and radiotherapy were distinctly lower than those before treatment(P <0.01), but were significantly increased in recurrence cancer patients (P <0.01). The soluble Apo-1/Fas levels in pleural and ascites fluid of malignant tumors were significantly higher than those in tuberculous effusions and transudates.</p><p><b>CONCLUSION</b>The soluble Apo-1/Fas levels in plasma, pleural and ascites fluid of malignant tumor patients are markedly increased, which might be associated with the progress of cancers. The changes of soluble Apo-1/Fas levels may be useful for understanding the pathologic process of cancers and to differential diagnosis of various pleural and ascites fluids.</p>

Detection and clinical significance of plasma urokinase-type plasminogen activator and its soluble receptor in patients with multiple myeloma / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the plasma levels of urokinase-type plasminogen activator(u-PA) and its soluble receptor(suPAR )in patients with multiple myeloma(MM) and to evaluate their clinical significance.</p><p><b>METHODS</b>Plasma u-PA and suPAR levels in 34 MM cases were measured with enzyme- linked immunosorbent assay (ELISA). The changes of plasma u-PA and suPAR levels in 6 MM cases were observed in succession before and after chemotherapy.</p><p><b>RESULT</b>The plasma u-PA and suPAR levels of MM patients were significantly higher than those of controls. The plasma u-PA and suPAR levels in the progress period was significantly higher than those in the stable period of MM patients as well as in controls (P<0.01), whereas there were no significant difference between the stable period of MM patients and controls (P>0.05). Among 6 cases,the plasma u-PA and suPAR levels after chemotherapy were significantly lower than those before chemotherapy (P<0.05). MM patients with tumor cells >20% in bone marrow smear had higher levels of plasma u-PA and suPAR than those with tumor cells <19% (P<0.05 P<0.01). The plasma u-PA and suPAR levels were positively correlated with the levels of serum globulin and the percentage of tumor cells in bone marrow,but negatively correlated with the levels of serum albumin.</p><p><b>CONCLUSION</b>Plasma u-PA and suPAR levels can serve as an index for clinical staging and assessing the therapeutic effect in MM patients.</p>

Relationship between acute organophosphorus pesticide poisoning and damages induced by free radicals / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the relationship between abnormal reactions of free radicals in bodies of patients with acute organophosphorus pesticide poisoning (AOPP) and damages induced by free radicals.</p><p><b>METHODS</b>58 AOPP patients and 58 healthy adult volunteers (HAV) were enrolled in an independent samples control design, in which spectrophotometric methods were used to determine the concentrations of nitric oxide (NO) and lipoperoxides (LPO) in plasma, and LPO in erythrocytes, vitamin C (VC), vitamin E (VE) and beta-carotene (beta-CAR) in plasma as well as activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and acetylcholinesterase (AChE) in erythrocytes.</p><p><b>RESULTS</b>Compared with the average values of every biochemical parameter in the HAV group, the average values of LPO in plasma and in erythrocytes, and NO in plasma in the AOPP group were significantly increased (P = 0.000001), while the average values of VC, VE, beta-CAR in plasma as well as SOD, CAT, GSH-Px and AChE in erythrocytes in the AOPP group were significantly decreased (P = 0.000001). The findings of Pearson product-moment correlation analysis between the value of AChE in erythrocytes and the values of above biochemical parameters for 58 AOPP patients showed that there was a significant linear negative correlation between AChE in erythrocytes and LPO, NO in plasma, and LPO in erythrocytes (P = 0.000001-0.001319), while there was a significant linear positive correlation between AChE in erythrocytes and VC, VE, beta-CAR in plasma as well as SOD, CAT, GSH-Px in erythrocytes (P = 0.000013-0.000824). The results of discriminant analysis of above chemical parameters for 58 AOPP patients and 58 HAV suggested that the correct rates of discriminant analysis were increased to 100% when the values of AChE and LPO in plasma and in erythrocytes, or AChE and others, were jointly used for the discriminant analysis.</p><p><b>CONCLUSION</b>The findings of the present study suggest that a series of free radical reactions in AOPP patients' bodies are pathologically aggravated, and the discriminant analysis used the above biochemical parameters could markedly increase its correct rates for AOPP patients.</p>