OBJECTIVE: To explore the effects of Ena/VASP gene family on the expression of glycoprotein (GP) Ib-IX complex in human megakaryoblastic leukemia Dami cells. METHODS: SiRNAs targeting Ena/VASP gene family were designed and synthesized to interfere Enah, EVL and VASP gene expression. When the siRNAs were transfected into Dami cells by using Lipofectamine^TM 2000 for 48 h, the expression of GPIb-IX complex was detected by quantitative real-time PCR, Western blot and flow cytometry. RESULTS: We successfully established siVASP , siEVL and si Enah Dami cell lines. And it was found that the expression of GPIb-IX complex had no evident reduction in siEVL or siVASP Dami cells at both mRNA and protein level, while the total protein and membrane protein of GPIb-IX complex were obviously reduced when Enah was knocked down. CONCLUSION Enah could affect the expression of GPIb-IX complex in human megakaryoblastic leukemia Dami cells, but the underlying mechanism still needs to be further explored.
Humans ; Cell Line ; Platelet Glycoprotein GPIb-IX Complex/metabolism* ; Leukemia/metabolism* ; Blood Platelets/metabolism*

OBJECTIVE: To investigate the hematopoietic protective effect of platelet-derived growth factor (PDGF)-BB on radiation-induced myelosuppression model mice and effect of anti-apoptosis of megakaryocyte line Meg-01 cells, and its possible mechanism. METHODS: Mice were radiated with 4 Gy of ¹³⁷Csγ ray to establish the model of myelosuppression. Mice were weighed and peripheral blood cell were counted before radiation (day 0) and day 7, 14 and 21 after radiation. On the 21 ^st day, the mice were killed. The sternal tissues of the mice were taken for morphological observation, and the femoral bone marrow cells were cultured for the assay of colony cell forming units (CFU). Meg-01 cells were cultured without FBS for 24 h to induce apoptosis, and then treated with PDGF-BB for 48 h. The effects of PDGF-BB on the proliferation were investigated by cell counting. Flow cytometry was used to detect early apoptosis (Annexin V), mitochondrial membrane potential (JC-1) and the expression of caspase-3. RESULTS: Peripheral blood cell counts of mice showed that PDGF-BB stimulated the recovery of white blood cells, red blood cells and platelets after radiation (P<0.05), especially for white blood cells. Morphological examination showed bone marrow hyperplasia in PDGF-BB group, the numbers of megakaryocytes and their progenitor cells were higher than those in the control group. PDGF-BB significantly stimulated the formation of CFU-MK, CFU-GM, BFU-E and CFU-F. PDGF-BB showed a strong proliferation effect in the concentration range of 5-50 ng/ml (P<0.001). PDGF-BB (50 ng/ml) significantly reduced the positive expression of Annexin V (P<0.01). The mitochondrial membrane potential in the control group was decreased when compared with PDGF-BB group, which indicated that the number of apoptotic cells was increased (P<0.01). Besides, the expression of caspase-3 in PDGF-BB group was significantly lower than that in control group (P<0.05). CONCLUSION PDGF-BB has a protective effect on the hematopoietic system of myelosuppression model mice, especially megakaryocytes and their progenitor cells. PDGF-BB has pro-proliferative and anti-apoptotic effects on Meg-01 cells, and the mechanism may be mediated through JC-1 and caspase-3 pathway.
Animals ; Mice ; Becaplermin ; Caspase 3 ; Hematopoietic System ; Apoptosis

OBJECTIVE: To investigate the apoptosis of CD34CD38-KG1a leukemia stem cells induced by Qinba selenium-mushroom extract(FA-2-b-β), and its related mechanism. METHODS: CD34CD38--KG1a cells were isolated from KG1a cell line by magnetic activated cell sorting. The proliferation ability of KG1a stem cells treatd by various concentration of FA-2-b-β(1.2-2.4 mg/ml) in vitro for 24 and 48 hours were tested by cell counting Kit-8(CCK8). Flow cytometry was used to detect the apoptosis rate of KG1a stem cells in each group after treated by FA-2-b-β in vitro. Expression of BAX,BCL-2,Casepase-3 and Cyclin D1 protein were detected by Western blot. RESULTS: The proportion of CD34CD38--KG1a stem cells was (95.35±2.63）% after immunomagnetic isolation. The proliferation of KG1a stem cells was inhibited significantly by FA-2-b-β, which shows a time- and dose-dependent manner (24 h,r=0.943; 48 h,r=0.976). Flow cytometry shows that with the increasing of drug concentration, the apoptosis was also increased, when KG1a stem cells was treated by FA-2-b-β for 24 h. Western blot indicated that the expression of apoptosis-related protein BAX and Casepase-3 were up-regulated, the expression of BCL-2 and Cyclin D1 were down-regulated. CONCLUSION FA-2-b-β can regulate proliferation and apoptosis KG1a stem cells, the involved mechanism may be related with the activation of mitochondrial-mediated apoptotic pathway.
ADP-ribosyl Cyclase 1 ; Antigens, CD34 ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Membrane Glycoproteins ; Neoplastic Stem Cells ; Selenium

OBJECTIVE: To determine the significance of morphology of bone marrow smear for diagnosis of bone marrow involvement in patients with diffuse large B-cell lymphoma (DLBCL), and to study the morphological characteristics of DLBCL cells involved in bone marrow. METHODS: Four hundred and twenty cases of DLBCL diagnosed at Peking Union Hospital from 2006 to 2016 were analyzed and identified. RESULTS: Blinded analysis of bone marrow smear and bone marrow biopsy data showed involvement in 42 cases on smears (S), in 47cases by biopsy (B) and the in 49 cases by (S+B). There was an excellent correlation between 2 methods diagnosing the bone marrow infiltration of DLBCL independently (κ=0.889). The morphological features of DLBCL cells involved in bone marrow were of medium sizes, round or irregular nuclear. The chromatin presented dark purple rea and coarse granular, and most of them had 1-5 nucleoli. The amount of cytoplasm was moderate with the color of dark blue or greyish blue. Vacuoles and pseudopodia were common. CONCLUSION The morphological examination of bone marrow cells has a certain role in the diagnosing bone marrow involvement in patients with DLBCL, and the atypical lymphoid cells making up ≥1% of the total nucleated cells highly suggests the bone marrow involvement in the patients with DLBCL.
Biopsy ; Bone Marrow ; Bone Marrow Cells ; Humans ; Lymphocytes ; Lymphoma, Large B-Cell, Diffuse ; Prognosis

OBJECTIVETo explore the role of bromodomain and extra terminal (BET) bromodomain in hematopoietic differentiation from human enbryonic stem cells (hESC).

METHODSThe effect of BET hematopoietic inhibitor I-BET151 on hematopoietic differentiation from hESC was detected by using a monolayer hematopoietic defferentiation model, immunofluorescence, flow cytometry and real-time PCR; moreover the role of I-BET151 in process of hematopoietic differentiation was explored by adding I-BET151 in different differentiation stages.

RESULTSThe analysis results of immunofluorescence, flow cytometry and real-time PCR showed that I-BET 151 significantly inhibited the generation of CD43 positive hematopoietic stem and progenitor cells (HSPCs). It was found that the addition of I-BET 151 in different stages, including APLNR lateral plate mesoderm production, CD34CD31 hemogenic endothelium (HEP) generation and endothelial-to-hematopoietic transition, significantly suppressed the generation of CD43 positive hematopoietic progenitor cells.

CONCLUSIONI-BET 151 inhibites hematopoietic differentiation from hESCs at several stages, suggesting that the BET bromodomain plays important roles in multiple stages of hematopoietic differentiation from hESCs.

OBJECTIVETo investigate the role of HSP90 in proliferation and apoptosis of leukemia cells K562 through detecting the effect of HSP90 inhibitors 17-[2-(Dimethylamino) ethyl] amino-17-desmethoxygeldanamycin(17-DMAG) on leukemia K562 cell lines.

METHODSThe K562 cells were treated with HSP90 inhibitors 17-DMAG, the semi-quantitative PCR was used to detect HSP90 gene expression, the WST was used to detect the effect 17-DMAG on cell proliferation as well as Annexin V flow cytometry was used to detect the cell apoptosis.

RESULTSAfter 17-DMAG treated the K562 cells in different stage, the K562 cell growth was obviously inhibited with time dependent (48 h)(r=0.9918) and dose dependent(3.2 µmol/L) manners (r=0.9999) (P<0.01); after the K562 cells in different stage were treated with different concentrations of 17-DMAG, the K562 cells showed significant apoptosis and with dosage-dependent mauner (r=0.9903)(P<0.01); HSP90 mRNA expression decreased significantly after K562 cells were treated with different concentrations of 17-DMAG for 48 hours. 17-DAMG down-regulated the HSP90 mRNA expression in dosage-dependent mauner as well(r=0.9227) (P<0.01).

CONCLUSIONHSP90 inhibitor 17-DMAG can inhibit the proliferation of K562 cells and induce their apoptosis. This study result provides laboratory basis for the treatment of leukemia patients with 17-DMAG.

OBJECTIVETo investigate the morphological manifestation of bone marrow cells in newly-diagnosed patients with POEMS syndrome.

METHODSThe bone marrow cells in 155 patients with POEMS syndrome were classified and counted by OLYMPUS BX51 microscope, and the abnormal morphology of bone marrow cells was observed.

RESULTSThe count of plasma cells with normal morphology was 83.9% (130/155), the count of plasma cells with abnormal morphology (< 5 percent) was 12.3% (19/155), the count of plasma cells with obvious abnormal morphology (> 10 percent) was 3.8% (6/155) in patients with POEMS syndrome.

CONCLUSIONThe morphology of plasma cells in the most patients with the POEMS syndrome are normal, the minor patients of the POEMS syndrome have little abnormal plasma cell morphology, the extremely few patients showed obvious morphological abnormality in the bone marrow plasma cells. The higher proportion of plasma cells, the more easily and more abnormal plasma cells will be found.

OBJECTIVETo construct a three-dimensional (3D) model of arteries supplying the extrahepatic bile duct with a new segmentation algorithm based on submillimeter CT data.

METHODSThe new image segmentation algorithm based on interactive volume rendering was integrated into Medical Image Three-Dimensional Visualization System (MI-3DVS) as an intersected plug-in. The abdominal submillimeter CTA data of 10 patients were imported into MI-3DVS and the 3D model of the extrahepatic bile duct and its supplying arteries were constructed. The 3D model was zoomed in, zoomed out and spinned for observation and analysis of the arteries supplying the extrahepatic bile duct.

RESULTSThe 3D models of the blood supply to extrahepatic bile duct allowed stereoscopic, and accurate display of the fourth- and fifth-level branches of the hepatic artery, the second-level branches of the cystic artery, the pancreatic duodenal artery arch and the retroportal artery. The 3D models also provided a clear vision of the biliary structures including the hepatobiliary tract, the left and right hepatic ducts, gallbladder, the liver duct, and the common bile duct.

CONCLUSIONBased on the segmentation method of interactive volume rendering, the CT data of the arterioles supplying the extrahepatic bile duct can be extracted and segmented for 3D reconstruction to display the three-dimensional anatomical structures of the extrahepatic bile duct and its supplying arteries.

Actins ; metabolism ; Animals ; Brain ; cytology ; metabolism ; Cell Culture Techniques ; methods ; Cells, Cultured ; Immunohistochemistry ; Pericytes ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; Staining and Labeling

Biomarkers, Tumor ; metabolism ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; Lymphoma ; genetics ; metabolism ; pathology ; Lymphoma, Follicular ; genetics ; metabolism ; pathology ; Lymphoma, T-Cell, Peripheral ; genetics ; metabolism ; pathology ; Signal Transduction ; Skin Neoplasms ; metabolism ; pathology ; T-Lymphocytes, Helper-Inducer ; metabolism ; pathology