PURPOSE: Prolonged fetal hypoxia stimulates erythropoiesis in fetal life and induces increased nucleated erythrocytes(NRBC) counts at the early newborn period. To evaluate the relationship between prolonged fetal hypoxia and neonatal intraventricular hemorrhage (IVH), and the prediction of neonatal IVH by neonatal NRBC. METHODS: We compared the daily courses of the absolute NRBC count in preterm new- boms at 34 weeks' gestation or earlier with(n=17) and without(n=20) IVH for 7 days of life. RESULTS: Absolute NRBC counts at birth were higher in neonates with IVH than in control neonates(2,499/mm3+/-3,748 and 412/mm3+/-272, respectively, P=0.0022). The cut-off value of 1,000/mm3 for absolute NRBC counts at birth showed the best parameter estimate of the predictive model for IVH at early newborn period with 100% of positive predictive value and 74.1% of negative predictive value. CONCLUSION: Prolonged fetal hypoxia inducing fetal erythropoiesis near labor is closely related to IVH at early newborn period. Thabsolute NRBC counts at birth is the very important predictable marker for the condition.
Erythroblasts* ; Erythropoiesis ; Fetal Hypoxia ; Hemorrhage* ; Humans ; Infant, Newborn* ; Parturition ; Pregnancy

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 <5% for neutrophils and lymphocytes, <15% for eosinophils and basophils and <7% for erythroblasts. The correlation coefficients (r) 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.
Basophils ; Eosinophils ; Erythroblasts ; Fluorescence ; Granulocytes ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils

OBJECTIVE: To explore the role of Ter cells in the development of the collagen-induced arthritis (CIA), we detected their quantity changes in the spleen of different stages of CIA mice and analyzed the correlation between Ter cells and the joint scores, and we also analyzed the correlation between Ter cells and the frequencies of T and B cell subsets, so as to further understand the pathogenesis of rheumatoid arthritis. METHODS: The six to eight weeks DBA/1 mice were used to prepare CIA model. After the second immunization, we began to evaluate the joint score. According to the time of CIA onset and the joint score, the CIA mice were divided into three stages: early, peak and late stages. According to the final joint score, the CIA mice at the peak stage were subdivided into the high score group (score>8) and the low score group (score≤8). The frequencies of Ter cells in the spleen of the naïve mice and the CIA mice at various stages and the frequencies of T and B cell subsets in the spleen of the CIA mice at the peak stage were detected by flow cytometry, then we carried on the correlation analysis. RESULTS: The frequencies of Ter cells in the spleen of the CIA mice was significantly higher than those of the naïve mice (8.522%±2.645% vs. 1.937%±0.725%, P<0.01), the frequencies of Ter cells in the spleen of the high score group mice was significantly lower than those of the low score group (6.217%±0.841% vs. 10.827%±0.917%, P<0.01). The frequencies of Th1 cells in the spleen of the high score group mice was significantly higher than those of the low score group mice (1.337%±0.110% vs. 0.727%±0.223%, P<0.05). The frequencies of Th17 cells in the spleen of the high score group mice was higher than those of the low score group mice (0.750%±0.171% vs. 0.477%±0.051%, P=0.099). The frequencies of germinal center B cells in the spleen of the high score group mice was significantly higher than those of the low score group mice (1.243%±0.057% vs. 1.097%±0.015%, P<0.05). Correlation analysis results showed that the frequencies of Ter cells in the spleen of the CIA mice at the peak stage was strongly negatively correlated with the frequencies of CD4+ T, Th1, Th17, and germinal center B cells, and was strongly positively correlated with the frequencies of B10 cells, indicating that these cells might have a protective effect in CIA. Studies on dynamic changes showed that the frequencies of Ter cells in the spleen of the CIA mice at the late stage was significantly lower than those at the peak stage (0.917%±0.588% vs. 8.522%±2.645%, P<0.001), suggesting the protective effect of these cells in arthritis. CONCLUSION Ter cells were significantly increased in the spleen of the CIA mice at peak stage, and were negatively correlated with joint scores and pathogenic immune cells, and positively correlated with protective immune cells. Ter cells were significantly decreased in the spleen of the CIA mice at the late stage. What we mentioned above suggests that Ter cells might be involved in the progression of rheumatoid arthritis as an immunomodulatory cell,but further in vivo and in vitro experiments are needed to verify its specific effects and mechanism.
Animals ; Arthritis, Experimental ; Erythroblasts ; Mice ; Mice, Inbred DBA ; Th17 Cells

We performed dual color fluorescence in situ hybridization (FISH) for the bcr/abl fusion in CML using the peripheral blood smears without destruction of cell morphology to determine the bcr/abl fusion. Two patients of CML, one patient in accelerated phase and one patient in chronic phase, were selected. The blood smears were fixed in absolute methanol. FISH was performed with the Mbcr/abl translocation DNA probe mixture and the slides were stained with Wright's stain after FISH. The blood smears of both cases revealed distinct signals without destruction of cellular morphology. The normoblasts and lymphocytes revealed beautiful fused bcr/abl signals as well as granulocytes in both cases. The results provide a novel finding that the normoblasts and lymphocytes in CML are also neoplastic clonal cells which has not been demonstrated with a single-cell approach before.
DNA ; Erythroblasts* ; Fluorescence* ; Granulocytes ; Humans ; In Situ Hybridization ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Lymphocytes* ; Methanol

The relationship between the kidney and erythropoiesis was investigated. Thirty-three rabbits were used of which nine rabbits were subjected to bilateral nephrectomy, eight to unilateral nephrectomy with ureter ligation of the oppositekidney and nine to bilateral ureter ligation. The controls consisted of four rabbits. In each group, N.P.N., B.U.N., creatinine, normoblast count and half time of plasma iron disappearance(Fe59) were measured 72 hours after the operation and the following data were obtained. Normal group:N.P.N.,27.9mg%(18.5-36.0mg%), B.U.N.,17.2mg%(9.9-30.4mg%), creatinine, 1.3mg%(0.7-1.62mg%)normoblast, 28%(13-36%) and half time of plasma iron disappearance, 62min(40-92min) Bilaterally nephrectomized group: N.P.N.,262.1mg%(198.0-267.3mg%), B.U.N.,141.2mg%(130.5-146.0mg%), creatinine, 19.1mg%(9.45-19.47mg%) ,normoblast ,22% (1.0-5.0%)and half time of plasma iron disappearance, 139.8min(87-196min) Uinlateral nephrectomized group with ureter ligation of the opposite kidney: N.P.N.,256.2mg%(199.8-420.0mg%), B.U.N.,108.0mg%(90.0-112.5mg%) ,creatinine, 17.4mg%(12.4-18.7mg%),normoblast, 14.2% (8.0-26.0%) and half time of plasma irondisappearance, 86.0min(51.0-159.0min) Bilaterally ureter ligated group: N.P.N.,242.7mg%(236.4-333.0mg%), B.U.N.,135.5mg% (129.0-151.3mg%) ,creatinine, 19.8mg%(18.9-21.3mg%), normoblast, 18.2% (13.0-20.0%) and half time of plasma iron disappearance, 69.5min(49.0-92.0min) Normoblast markedly decreased(to 2.2%) and half time of plasma iron disappearance(Fe59) was strikingly higher(87 to 196 min., mean 139,8min.)than normal rabbits(40 to 92 min., mean 62 min.) in bilateral nephrectomized group. This marked depression of erythropoiesis occurred in bilateral nephrectomized group while in the group of bilateral ureterligation and unilateral nephrectomy with ureter ligation of opposite kidney only moderate depression of erythropoiesis was noted despite the elevation in azotemia and the degree of malnutrition were almost same level in these three groups. It suggests that the kidney is probably the source of and erythropoietic factor.
Azotemia ; Creatinine ; Depression ; Erythroblasts ; Erythropoiesis* ; Iron ; Kidney* ; Ligation ; Malnutrition ; Nephrectomy ; Plasma ; Rabbits* ; Ureter

OBJECTIVE: In an attempt to further maximize the potential of genetic analysis from fetal cells isolation, fetal nucleated red blood cell (FNRBC) recovery with direct anti-gamma hemoglobin staining after density gradient and depletion was compared with three different whole blood magnetic separations (1-step and 2-step ferrofluid, 2-step Dynal beads). METHODS: In model systems such as quantitatively defined spikes of fetal into adult blood, as well as blood samples after surgical termination procedures, fetal cell yield and purity through the results of fluorescence in situ hybridization (FISH), quantitative real time polymerase chain reaction (PCR), and fluorescence-activated cell sorting (FACS) were calculated. RESULTS: The yield of total number of cells with a XY signal after FISH was the highest on direct anti-gamma hemoglobin staining. After normalizing the results of each experiment to the corresponding result from anti-gamma hemoglobin staining (1), ratio is 0.42 in 1-step ferrofluid, 0.33 in 2-step ferrofluid, and 0.76 in 2-step dynal beads. The fetal cell purity is clearly better in direct anti-gamma hemoglobin staining than those of the magnetic separations from whole blood. The median ratio is 56.3% in anti-gamma hemoglobin staining, 7.7% in 1-step ferrofluid, 6.5% in 2-step ferrofluid, and 31.4% in 2-step dynal beads. CONCLUSION: This study shows that the direct anti-gamma staining is the best fetal cell recovery system and it is very useful to isolate fetal nucleated red blood cells as a non-invasive genetic source.
Adult ; Antibodies* ; Erythroblasts* ; Erythrocytes ; Flow Cytometry ; Fluorescence ; Humans ; In Situ Hybridization ; Real-Time Polymerase Chain Reaction

OBJECTIVE: To evaluate the intrauterine hypoxic effect in term small-for-date (SGA) neonates by comparing the umbilical venous erythropoietin (EPO) concentration between appropriately-grown (AGA) and SGA neonates at delivery and to determine the variables that correlate with the umbilical venous EPO concentration by multiple regression analysis. METHODS: 183 term singleton neonates (gestational weeks > OR =37) were enrolled and divided into 136 cases of AGA (10th-90th percentile of birth weight for the gestational age) and 47 cases of SGA (< 10th percentile of birth weight for the gestational age. At each delivery, blood gas values, concentration of EPO by radioimmunoassay and the number of nucleated erythrocytes (NRBC) per 100 white blood cells in smear of umbilical venous blood were obtained. The placentas were examined microscopically for presence of pathological infarct. Statistical analysis was done by Mann-Whitney U test, x2 test, and univariate and multiple regression analysis using SPSS statistical package (version 10). RESULTS: The median umbilical venous EPO concentration, fetal hemoglobin level were significantly higher in SGA neonates than those in AGA neonates. There was no difference in number of NRBC between AGA and SGA neonates. Multiple regression analysis model with level of NRBC, presence of placental infarct and SGA provided prediction of EPO level in umbilical venous blood at delivery (regression equation: EPO=-103.94+4.75NRBC+68.07placental infarct+36.40SGA F=15.57. r2=0.47). CONCLUSION: Term SGA neonates was considered to have compensatory, not pathological intrauterine hypoxic effect by showing increased level of EPO and normal level of NRBC in umbilical venous blood at delivery, compared with thoses of AGA. In the suspected cases of SGA antenatally, measurement of NRBC level and placental pathologic examination for infarct can be informative for estimating the extent and duration of intrauterine hypoxia.
Anoxia ; Birth Weight ; Erythroblasts ; Erythropoietin* ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn* ; Leukocytes ; Placenta ; Pregnancy* ; Radioimmunoassay

OBJECTIVE: To evaluate the intrauterine hypoxic effect in term small-for-date (SGA) neonates by comparing the umbilical venous erythropoietin (EPO) concentration between appropriately-grown (AGA) and SGA neonates at delivery and to determine the variables that correlate with the umbilical venous EPO concentration by multiple regression analysis. METHODS: 183 term singleton neonates (gestational weeks > OR =37) were enrolled and divided into 136 cases of AGA (10th-90th percentile of birth weight for the gestational age) and 47 cases of SGA (< 10th percentile of birth weight for the gestational age. At each delivery, blood gas values, concentration of EPO by radioimmunoassay and the number of nucleated erythrocytes (NRBC) per 100 white blood cells in smear of umbilical venous blood were obtained. The placentas were examined microscopically for presence of pathological infarct. Statistical analysis was done by Mann-Whitney U test, x2 test, and univariate and multiple regression analysis using SPSS statistical package (version 10). RESULTS: The median umbilical venous EPO concentration, fetal hemoglobin level were significantly higher in SGA neonates than those in AGA neonates. There was no difference in number of NRBC between AGA and SGA neonates. Multiple regression analysis model with level of NRBC, presence of placental infarct and SGA provided prediction of EPO level in umbilical venous blood at delivery (regression equation: EPO=-103.94+4.75NRBC+68.07placental infarct+36.40SGA F=15.57. r2=0.47). CONCLUSION: Term SGA neonates was considered to have compensatory, not pathological intrauterine hypoxic effect by showing increased level of EPO and normal level of NRBC in umbilical venous blood at delivery, compared with thoses of AGA. In the suspected cases of SGA antenatally, measurement of NRBC level and placental pathologic examination for infarct can be informative for estimating the extent and duration of intrauterine hypoxia.
Anoxia ; Birth Weight ; Erythroblasts ; Erythropoietin* ; Fetal Hemoglobin ; Gestational Age ; Humans ; Infant, Newborn* ; Leukocytes ; Placenta ; Pregnancy* ; Radioimmunoassay

The isolation of fetal cells from maternal circulation has the potential to allow relativelyself prenatal diagosis for all pregnant women. The present technology, however, has notreached the accuracy required for clinical diagnosis because of maternal cell contaminationSo we published a new method for enrichment of nRBC in a fetal cell isolation(1996).In this study, attempted to FISH analysis of nRBC which was isolated by our ownmethods. We evaluated the efficiency of FISH.As the results, we have successfully used FISH on enriched nRBC.We were able to identified 2 abnormal fetus which were confirmed by conventionalcytogenentic study as Down syndrome(Fig.1) and Klinefeltre syndrome(Fig.2). And thesensitivity and specificity for FISH was 86%(49/57) and 92.3%(36/39), respectively.According to our results, fetal cell analysis by FISH can be reliable used for prenatalaneuploidy diagnosis. However, the problems of enrichment of the fetal cell and FISH probeor condition should be over come before analyze.
Aneuploidy ; Diagnosis ; Erythroblasts* ; Female ; Fetus ; Fluorescence* ; Humans ; Pregnant Women ; Sensitivity and Specificity

The primitive erythroblasts in 21 cases of embryonic hearts from 4 to 9 weeks of gestation were studied with a light microscope. The nuclear diameter, the motosis, and the loss of nuclei of the primitive erythoblasts were analyzed quantitatively. The results obtained were as follows. 1. At 4 weeks of gestation, the blood cells consisted of proerythroblasts, along with basophilic polychromatophilic primitive erythroblasts. The nuclear diameter ranged from 3.20 µm to 9.20 µm, but the main range was from 4.20 µm to 6.00 µm. It was revealed that 9.50% had diameter of more than 6 µm. 2. At the fist half of the 7 week gestation when hepatic hemopoiesis developed, the blood cells consisted of basophilic, polychromatophilic, and eosinophilic erythroblasts. Cells of more than 6 µm in nuclear diameter were about 1.10% and thereafter gradually disappeared. The range of the nuclear diameters was from 2.60 µm to 7.00 µm, while a range from 3.40 µm to 5.20 µm wqs the main. The proportion of cells less than 4 µm in nuclear diameter was 39.58% and thereafter rapidly increased. 3. From the second half of 7 weeks to 9 weeks of gestation, the erythrocytes originating from hepatic hemopoiesis increasingly replaced the circulating primitive erythroblasts, which became mature during this time. The erythrocytes showed 72.88% at 9 weeks of gestation. The proportions of cells less than 4 µm in nuclear diameter in the first and second haIves of 8 weeks and 9 weeks were 52.73%, 80.02%, and 89.09%, which represented the rapid destruction of nuclei. 4. Mitosis in the primitive erythroblasts occurred principally up to the early 6th weeks, and very weakly at 8 weeks. 5. As the crown-rump length increased, the average nuclear diameter decreased very significantly (P<0.01, y=-0.2811X + 0.3171). The results suggest that distrilbution of the nuclear diameter, the maturity, the rate of nuclear loss, and the mitotic figure offer credible data for estimating embryonic age.
Basophils ; Blood Cells ; Crown-Rump Length ; Embryonic Structures* ; Eosinophils ; Erythroblasts* ; Erythrocytes ; Heart ; Humans* ; Mitosis ; Pregnancy