Angelica sinensis polysaccharide promotes stress erythropoiesis in mice caused by 5-FU / 中国药理学通报

Aim To investigate the effects of angelica sinensis polysaccharide (ASP) antagonizing 5-fluorou-raeil (5-FU) on spleen stress erythropoiesis in mice and its related mechanism. Methods C57BL/6J mice aged 6-8 weeks were randomly divided into control group, ASP group, 5-FU group and ASP + 5-FU group. The mouse body weight during the modeling pe-riod was recorded, and peripheral blood routine and the number of mononuclear cells in the bone marrow of femur were measured. Histopathology of spleen was de-tected, also the index and cellularity of spleen were analyzed. BFU-E of spleen mononuclear cells was counted. The number of F4/80

Prediction of the anti-inflammatory mechanisms of curcumin by module-based protein interaction network analysis

Curcumin, the medically active component from Curcuma longa (Turmeric), is widely used to treat inflammatory diseases. Protein interaction network (PIN) analysis was used to predict its mechanisms of molecular action. Targets of curcumin were obtained based on ChEMBL and STITCH databases. Protein-protein interactions (PPIs) were extracted from the String database. The PIN of curcumin was constructed by Cytoscape and the function modules identified by gene ontology (GO) enrichment analysis based on molecular complex detection (MCODE). A PIN of curcumin with 482 nodes and 1688 interactions was constructed, which has scale-free, small world and modular properties. Based on analysis of these function modules, the mechanism of curcumin is proposed. Two modules were found to be intimately associated with inflammation. With function modules analysis, the anti-inflammatory effects of curcumin were related to SMAD, ERG and mediation by the TLR family. TLR9 may be a potential target of curcumin to treat inflammation.

Evaluation of the Mindray BC-6800 Complete Blood Counts Analyzer

BACKGROUND: The BC-6800 (Mindray, China) is a recently developed hematology analyzer that utilizes 'SF Cube Technology' to improve the reliability of complete blood counts (CBC), white blood cell (WBC) differentials, and erythroblast counts. In this study, we evaluated the performance of the BC-6800 for CBC, WBC differentials, reticulocyte counts, and erythroblast counts and analyzed the efficiency of its flag system. METHODS: Specimens from 100 healthy controls and 95 patients were used. We performed precision and correlation studies of CBC, WBC differentials, reticulocyte counts, and erythroblast counts. We also analyzed the efficiency of the flag system in detecting abnormal blood cells. RESULTS: The coefficients of variation (CVs) of precision were 0.9800 for CBC except erythrocyte indices, and >0.9500 for WBC differentials except monocyte and basophil. The WBC differentials and erythroblast counts obtained using the BC-6800 were well correlated with those of manual counts. The efficiencies of the flag system were 77.9% for Blasts, 82.1% for Immature Gran, 86.3% for Atypical Lymph, and 92.6% for NRBC present. CONCLUSIONS: The BC-6800 showed good precision and correlation with pre-existing hematology analyzers. The flag systems were quite efficient for detecting abnormal blood cells. Our study demonstrated that the BC-6800 hematology analyzer exhibits suitable performance and is helpful in routine laboratories.

Evaluation of the ABX Pentra DX 120 for the Detection of Immature Cells in Peripheral Blood / 임상검사와정도관리

INTRODUCTION: The ABX Pentra DX 120 (Pentra DX 120, ABX Diagnostics, Montpellier, France) adopted new technologies to perform differential leukocyte and erythroblast counts. The double matrix can discriminate Large Immature Cell (LIC), Immature Granulocyte (IMG), Immature Monocyte (IMM), Immature Lymphocyte (IML), and Atypical lymphocyte (ALY) in addition to a routine 5-differential count. For erythroblast (ERB), a fluorescence method is employed. In this study, we evaluated the performance of the Pentra DX 120 in the performance of differential leukocyte and erythroblast counts. METHODS: Precision was evaluated using 3-level control materials. Comparison analysis was performed on 200 samples: 100 normal and 100 abnormal samples. We evaluated the 5 part differential count, LIC, IMG, IMM, IML, ALY, and ERB. These parameters were analyzed in comparison with the results from the reference method, manual differential count. RESULTS: The coefficients of variation (CVs) of precision were 0.9 except monocytes and basophils. IMG, ALY and erythroblasts were also well correlated with manual count (r=0.8315, 0.5602, 0.8144, respectively). The efficiency of flagging system was 84% for LIC, 80% for ALY, and 78.0% for increased ERB (>2/100WBCs). CONCLUSIONS: The Pentra DX 120 performed reliable differential leukocyte and IMG, ALY, and ERB results demonstrated comparable performance to manual count. And, the flagging system was efficient for detecting each abnormal cell population. We expect the Pentra DX 120 double matrix and erythroblast count can reduce microscopic review rate in routine laboratory and promote laboratory efficiency.

The utility of an erythroblast scoring system and gender-independent short tandem repeat (STR) analysis for the detection of aneuploid fetal cells in maternal blood / 대한산부인과학회지

OBJECTIVE: The aim of this study was to determine whether fetal nucleated red blood cells (NRBCs) could be distinguished from maternal cells in peripheral blood using an erythroblast scoring system. Presumptive fetal NRBCs were further analyzed through the use of fluorescent PCR amplification with polymorphic STR markers to prove fetal origin. METHODS: NRBCs were isolated by density gradient separation, CD15/45 depletion, and gamma hemoglobin positive selection from peripheral blood of seven women who had undergone termination of pregnancy because of fetal trisomy 21 (n=4), 18 (n=1), and 13 (n=2). Candidate fetal NRBCs, based on four discrete morphological and hemoglobin staining criteria, were then subjected to fluorescent PCR amplification of chromosome 21 short tandem repeat (STR) markers (D21S1411, D21S11) and chromosome 18 STR markers (D18S535). RESULTS: In all cases candidate fetal NRBCs were accurately identified based on erythroblast scoring system and confirmed to be fetal in origin based on the presence of shared and non-shared polymorphic DNA alleles when compared to DNA isolated from maternal cells. Also in five cases aneuploid fetal cells in maternal blood were identified through the use of fluorescent PCR amplification with polymorphic STR markers. CONCLUSION: We were able to distinguish fetal NRBCs from maternal cells and prove fetal origin independent of gender. These results suggest that this novel combined approach to fetal cell isolation through using an erythroblast scoring system and genetic analysis by STR analysis is a promising method for noninvasive prenatal diagnostic applications.

The examination of bone marrow erythroblastic islands and it′s clinical value / 中华检验医学杂志

Objective To investigate the correlation between the morphological features of erythroblastic islands (EI) and clinical outcome. Methods The incidence and maturity of EI were measured by morphological examination.Results The incidence of EI was highest in acute erythremia, acute erythroleukemia, and iron deficiency anemia (75.0%?70.0%?63.6%, respectively), while very rare in aplastic anemia, acute leukemia (acute erythroleukemia excepted ), lymphoma. In addition, EI incidence in leukemia was significantly higher after remission than at presentation (43.7% vs 0, P

Migration of Emperipoletic Erythroblasts Within Kupffer Cells in Human Hepatic Hemopoiesis: Electron Microscopic Study / 대한해부학회지

The presence of erythroblasts within Kupffer cell was studied for transmission electron microscopically with 5 human fetal livers from 11 to 23 weeks of gestation during the high activity of hepatic hemopoiesis. By using continuous series of thin sections electron microscopically, the objective of the present study was to evaluate the relevance between a migrated erythroblast and a Kupffer cell, and the migration of erythroblasts within Kupffer cells in the sinusoidal lumen. During the examined period the sinusoidal wall consisted of endothelial cells and Kupffer cells, being deficient in basement membrane. Erythropoietic cell-Kupffer cell interaction was often found as the emperipolesis and adhesion between the cells in human fetal liver under electron microscopy. The cytoplasm of the emperipoletic Kupffer cell contained several mitochondria, rough endoplasmic reticuli, clear vesicles, electron dense bodies, cellular debris with shrunken chromatin of enucleated nuclei, intact enucleated nuclei, and erythroblast bearing vacuoles as intact erythroblasts. Intracellular erythroblasts in the Kupffer cell remain unaltered with their normal structure and showed mitosis, enucleation and migration of erythroblast into the sinusoidal lumen. And a clear zone of a vacuole was readily seen around the intracellular erythroblast within Kupffer cell. On occasion, the hypertropic Kupffer cell with interacellular erythroblasts virtually occluded the sinusoidal lining cell. Processing of a migrating emperipoletic erythroblast within a Kupffer cell, the erythroblast migrated via migration pore through the luminal cell membrane of the Kupffer cell into the sinusoidal lumen. An invasion of a proerythroblast into Kupffer cell or a migration of the cell into the sinusoidal lumen had been found in human fetal liver from 11 to 13 weeks of gestation. The results demonstrate that migration of emperipoletic erythroblasts within Kupffer cells occurs in human hepatic hemopoiesis. We suggest that emperipolsis may be one of the mechanisms that support the maturation of erythroblasts in human fetal liver.

Maturation of Erythroblasts in Human Liver during Ontogeny - An Electron Microscopic Study / 대한해부학회지

The hepatic hemopoiesis in intrauterine lifeis predominently erythropoiesis,and the erythroblasts undergo the differentiation process finally to form reticulocytes.In this syudy, the development of erythropoiesis in human fetal liver was observed using transmission electron microscope. The immature erythroblasts were more prominent in earlier fetal liver and proliferated rather than went through final differentiation process. The erythroblasts of different differentiation stages exhibited apoptosis in addition to the normal differentiation process. The nuclei of acidophilic erythroblasts were removed by the excessive condensation and dissociation of nuclei from the cytoplasm or by the displacement of nuclei to one side of the cell with deformation of nuclei. The deformed nuclei restored the round shape after the completion of enucleation and engulfed by hepatocytes and Kupffer cells. Two types of erythroblast islands were present in hepatic plate by the differentiation stages of erythroblasts, id est, islands of the same and the different differentiation stages. And several erythroblasts and enucleated nuclei were included in hepatocytes, intrahepatic macrophages and Kupffer cells, and the intrahepatic macrophages resembling Kupffer cell could be suggested to be originated from the Kupffer cell. But there was no morphological evidence of phagocytosis of erythroblasts and nuclei by these cells. In summary, human fetal hepatic erythropoiesis occurred by forming erythroblastic islands and some erythroblasts proceeded to apoptosis during the differentiation. Hepatocytes and macrophages were present in close relation to erythroblast islands and were suggested to influence the development and differentiation of erythroblasts.

A HIGH EFFICIENT METHOD FOR THE SEPARATION OF CELL FRACTION ENRICHED WITH INTERMEDIATE AND LATE ERYTHROBLAST CELLS / 解剖学报

A method described by Harrison has been adopted and modified by us for the separation of intermediate and late erythroblast cells from 15 day embryonic liver of pregnant Wistar rat. The method consisted briefly of preparation of fetal liver cell suspension and the separation of cell types in a 40% and 70% nonlinear Percoll gradient system. Using this method, we can obtain about 96% of hemogenous population of intact and viable intermediate and late erythroblasts. Examination of tho separated cells by Giemsa and benzidine staining and by electron microscopic observation indicated that no granulocytes or other white blood cells could be detected, except for some contamination of about 1% of proerythroblasts and 3% of reticulocytes in the fraction. Trypan blue vital staining demonstrated that there were over 95％ of the cells maintained viable after separation, they could be used directly for the study of cell differentiation as well as biochemical analysis. SO, this is an economical, simple and easy technique to operate which proved to be a useful mean for obtaining enrich population of intermediate and late erythroblasts in research in the field of cell biology.

DETECTION OF GENE EXPRESSION PATTERNS IN HYBRIDS CROSSED BETWEEN RAT NUCLEATED ERYTHROBLASTS AND MOUSE MYELOMA SP2/0 CELLS USIND IN SITU HYBRIDIZATION TECHNIQUE / 解剖学报

The gene expression of both the mouse plasmocytoma (SP2/0) and the hybrid cells crossed with rat nucleated erythroblasts were detected by in situ hybridization technique using the probes of mouse ?-globin gene and 7 oncogenes (v-Ki-ras, v-H-ras, v-sis, erb-B, v-abl, v-fos, c-myc). After plasmic amplification, DNA was isolated by alkali lysis, purified and recovered, the DNA containing gene fragments were labelled with ~(32)P to become high activity ~(32)P-cDNA probes through nick translation, and the labelled probes were used to detect the gene transcripts in cellular level. The results indicated that: (1) no mouse ?-globin gene transcripts could be detected in the cytoplasm of SP2/0 cells, as well as in hybrid cells within 72 hours after cell fusion, but transcript signals could be observed in hybrid cells from 4th to 26th passages. (2) Active expression of multioncogenes in SP2/0 cells was demonstrated, all the 7 oneogenes tested, except v-sis, were expressed more strongly. On the other hand, the expression of oncogenes in hybrid cells was found to be dramatically decreased, among them, the oncogenes of c-myc and Ki-ras been suppressed completely. After long term of passages (26th subcultures), the expression of c-myc and Ki-ras was still lower than that of SP2/o ceils although in some cases other oncogenes increased in their expression levels. These results confirmed that the multistep carcinogenesis involved multi-oncogenes expression and that the decancerization of tumor cells may be due to the suppression of multi-oncogenes activity as well as to activate the expression of differentiation genes.

LIGHT AND ELECTRON MICROSCOPIC STUDIES ON HYBRIDS CROSSED BETWEEN RAT INTERMEDIATE OR LATE ERYTHROBLASTS AND MOUSE SP2/0 CELL LINE / 解剖学报

The present study reported the observations with light and electronic micros copy on hybrid cells crossed between rat intermediate or late erythroblasts and mouse SP2/0 plasmocytoma cells. In a short period after fusion, the cell size and the ratio of nuclear heterochromatin in hybrid cells appeared to be increased, but the number of nucleoli, as well as the number of microvilli, finger-like processes, and nuclear cytoplasmic ratio were decreased. Swelling mitochondria, pycnotic nuclei and/or process of enucleation also could be seen in some hybrid cells. The subcul tured hybrid cells were characterized with less microvilli and cellular surface membrane processes, and showing marked changes in nuclear size, as well as the appearance of cytoplasmic vesicles and dense granules in some cells. The observations mentioned above provide morphological and ultrastructural evidences for the regulation of malignant phenotype of hybrid cell model we reported previously. The possible relationship between the deeancerization and the morphological changes of hybrid cell nucleus, cytoplasm and cell surface were briefly discussed.