Characteristics of bone marrow compensatory erythropoiesis in hereditary spherocytosis / 中华血液学杂志

Objective: To reveal the compensatory features of bone marrow (BM) erythropoiesis in hereditary spherocytosis (HS) and to explore the effect of diferent hemoglobin levels on this compensation. Methods: Clinical and laboratory data of patients with HS were collected, and the peripheral blood absolute reticulocytes counts value was taken as the surrogate parameter to evaluate the ability of erythropoiesis compensation. BM erythropoiesis compensation in HS with diferent degrees of anemia were evaluated. Results: ①Three hundred and two patients were enrolled, including 115 with compensated hemolytic disease, 74 with mild anemia, 90 with moderate anemia, and 23 with severe anemia. ②Hemoglobin (HGB) was negatively correlated with serum erythropoietin in the decompensated hemolytic anemia group (EPO; rs=-0.585, P<0.001) . ③The median absolute reticulocyte count (ARC) of HS patients was 0.34 (0.27, 0.44) ×10(12)/L, up to 4.25 times that of normal people. The maximum ARC was 0.81×10(12)/L, about 10 times that of normal people. The median ARC of patients with compensated hemolytic disease was 0.29 (0.22, 0.38) ×10(12)/L, up to 3.63 times that of normal people. The median ARC of patients with hemolytic anemia was 0.38 (0.30, 0.46) ×10(12)/L, which was significantly higher than the patients with compensated hemolytic disease, up to 4.75 times that of normal people (z=4.999, P=0.003) . ④ ARC was negatively correlated with HGB in the compensated hemolytic disease group (rs=-0.177, P=0.002) and positively correlated with HGB in the decompensated hemolytic anemia group (rs=0.191, P=0.009) . There was no significant difference in the ARC among patients with mild, moderate, and severe anemia (χ(2)=4.588, P=0.101) . ⑤The median immature reticulocyte production index of the mild, moderate, and severe anemia groups was 13.1% (9.1%, 18.4%) , 17.0% (13.4%, 20.8%) , and 17.8% (14.6%, 21.8%) , respectively; the mild anemia group had lower index values than the moderate and severe anemia groups (P(adj) values were both<0.05) , but there was no significant difference between the latter groups (P(adj)=1.000) . The median immature reticulocyte count of patients in the mild, moderate, and severe groups was 5.09 (2.60, 7.74) ×10(10)/L, 6.24 (4.34, 8.83) ×10(10)/L, and 7.00 (3.07, 8.22) ×10(10)/L, respectively; there was no significant difference among the groups (χ(2)=3.081, P=0.214) . Conclusion: HGB can be maintained at a normal level through bone marrow erythropoiesis, while red blood cells are reduced in HS. However, once anemia develops, the bone marrow exerts its maximum erythropoiesis capacity and does not increase, regardless of anemia aggravation or serum EPO increase.

Abnormal expression of b10 cell frequencies: possible relation to pathogenesis and disease severity of aplastic anemia

SUMMARY OBJECTIVE: Aplastic anemia (AA) is an immune-mediated disease that destroys hematopoietic cells through activated T lymphocytes. B lymphocyte-mediated humoral immunity also plays an important role in the pathogenesis of AA. Regulatory B cell (Breg) subpopulation, which is defined as "B10", secretes interleukin 10 (IL-10). The objective of our experiment was to investigate whether the scale-down proportion of B10 cells in AA patients may play a key role in the pathogenesis. METHODS: A total of 38 AA patients (14 SAA patients and 24 NSAA patients) and 20 healthy control subjects were included. All subjects did not suffer from autoimmune diseases or any other diseases affecting the immune system, such as infectious diseases. Bone marrow mononuclear cells (PBMCs) were isolated and analyzed by Flow cytometry (FCM) and Immunofluorescence double-labeling assay. The relationship between the relative proportions of B10 and ProB10 and their associations to AA, as well as disease severity, were assessed by common clinical indicators and then examined. RESULTS: Our analyses revealed AA patients had significantly lower proportions of peripheral B10 and B10pro compared to healthy controls. SAA patients had a substantially lower percentage of B10 cells and B10pro cells compared to NSAA patients. In addition, B10 cells and B10pro cells were negatively correlated with absolute neutrophil counts, hemoglobin levels and platelet, and absolute reticulocyte counts in AA patients. CONCLUSIONS: The present study attempted to elucidate the potential role of the scale-down proportion of B10 cells in the pathogenesis of AA.

RESUMO OBJETIVO: A anemia aplástica (AA) é uma doença imunomediada que destrói células hematopoiéticas por meio dos linfócitos T ativados. A imunidade humoral mediada por linfócitos B também desempenha um papel importante na patogênese da AA. A subpopulação de células B reguladoras (Breg), que é definida como "B10", secreta interleucina 10 (IL-10). No experimento, investigou-se se a proporção reduzida de células B10 nos pacientes de AA pode desempenhar um papel-chave na patogênese. MÉTODOS: Um total de 38 pacientes de AA (14 pacientes de anemia aplástica grave e 24 pacientes de anemia aplástica não grave) e 20 indivíduos de controle saudáveis foram incluídos. Todos os indivíduos não sofriam de doenças autoimunes ou de quaisquer outras doenças que afetam o sistema imunológico, tais como doenças contagiosas. As células mononucleares da medula óssea (PBMCs) eram isoladas e analisadas por citometria de fluxo (FCM) e ensaio de dupla marcação por imunofluorescência. A relação entre as proporções relativas de células B10 e as células ProB10 e as suas associações à AA, assim como a gravidade da doença avaliada por indicadores clínicos comuns, foram examinadas. RESULTADOS: Nossas análises revelaram que os pacientes de AA têm proporções significativamente menores de células B10 e células ProB10 periféricas em comparação com indivíduos de controle saudáveis. Os pacientes de anemia aplástica grave tiveram uma percentagem substancialmente menor de células B10 e células B10pro em comparação com pacientes de anemia aplástica não grave. Além disso, as células B10 e B10pro foram negativamente correlacionadas com contagens absolutas de neutrófilos, níveis de hemoglobina e plaquetas e contagem de reticulócitos absolutos nos pacientes de AA. CONCLUSÕES: Além disso, o estudo presente tentou elucidar o papel imunorregulatório potencial das células B10 na patogênese da AA e fornecer uma nova estratégia para a aplicação de imunoterapia baseada na célula B para tratar a AA no futuro.

Correlação entre as contagens de reticulócitos manual e automática em amostras de felinos anêmicos / Correlation between manual and automatized values of reticulocyte count in anemic feline samples

O objetivo do presente estudo foi correlacionar os valores de reticulócitos pontilhados e agregados obtidos por metodologia manual com a metodologia automática de contagem de reticulócitos totais em amostras de sangue de gatos anêmicos, analisados em um contador hematológico com citometria de fluxo. Para isso, 40 amostras de sangue de pacientes felinos anêmicos, independentemente de idade e sexo, foram utilizadas para a determinação das contagens absolutas de reticulócitos totais pela metodologia automatizada por citometria de fluxo fluorescente e pela técnica manual com corante supravital, em duplicata. Na contagem manual, houve a discriminação entre reticulócitos pontilhados e agregados. Para a correlação entre os métodos, foi realizada a análise de regressão de Passing-Bablok. A média do hematócrito dos gatos foi de 15,25%, tendo a maioria dos gatos (32,5%) apresentado anemia moderada (hematócrito = 17,81%). Como resultados, a análise de regressão demonstrou que a correlação entre a contagem absoluta total automática foi superior à contagem manual de reticulócitos agregados (rho= 0,71; P<0,001) do que a contagem absoluta de reticulócitos pontilhados (rho= 0,68; P<0,001). Os resultados apresentados sugerem que a contagem de reticulócitos total absoluta realizada pelo analisador hematológico ProCyte Dx em gatos anêmicos se refere à contagem absoluta de reticulócitos. Dessa maneira, recomenda-se que os valores possam ser utilizados para a avaliação imediata da condição hematológica de gatos anêmicos.(AU)

The aim of this study was to correlate the punctate and aggregated reticulocytes values obtained by manual methodology and the automatic reticulocyte count in 40 blood samples from anemic cats. Total reticulocyte absolute counts were determined by automated fluorescence flow cytometry and manual methods in 40 blood samples obtained from anemic cats. The manual count was obtained by supravital stain in duplicate to each sample and the reticulocyte morphology were discriminated between punctate and aggregates reticulocytes. Passing-Bablok regression analysis was utilized to compare the methods. Most samples were from anemic cat (15,25%) and the hematocrit mean was 17,81%. Regression analysis showed that the correlation between the absolute total automatic counts is higher with aggregated reticulocytes (rho= 0,71; P< 0,001) than with absolute punctate reticulocytes counts (rho= 0, 68, P< 0.001). Results suggest that the ProCyte Dx reticulocytes count in anemic cats is correlated with aggregate reticulocyte count. Thus, the greater amount of RNA and organelles in aggregate reticulocytes generates a cellular complexity and, therefore, greater impregnation of the dye in an automatic count. Thus, the values obtained by the hematologic instrument can be used for the immediate evaluation of the hematological condition in anemic cats.(AU)

Contagem de Reticulócitos na Prática Clínica: um Exame Pouco Utilizado / Reticulocytes Counting in Clinical Practice: a Little Examination Used

Objetivo: Estimar o percentual de contagem de reticulócitos em relação aos hemogramas realizados nos laboratórios de análises clínicas de Divinópolis/MG. Métodos: Trata-se de um estudo descritivo realizado nos laboratórios de análises clínicas do município de Divinópolis/MG inscritos no Cadastro Nacional de Estabelecimento de Saúde (CNES). Foi realizada uma entrevista através de questionário a fim de obter as informações dos números de hemogramas e contagens de reticulócitos realizados durante os últimos 12 meses, do atendimento (ambulatorial ou hospitalar), da instituição (pública ou privada), do porte (pequeno, médio ou grande) e de quais os métodos eram utilizados para realização dos exames. Resultados: Dos 15 laboratórios inscritos no CNES, 11 foram entrevistados (73%). Foram realizados 276.666 hemogramas e 4.248 contagens de reticulócitos nos últimos 12 meses em todos os laboratórios, o que significa o percentual de 1,5% de contagem de reticulócitos em relação aos hemogramas realizados. A maioria dos laboratórios é privado (81%), de médio porte (54%) e realizam atendimento ambulatorial (54%). Para realizar o hemograma, a maioria (81%) utiliza a técnica citometria de fluxo e para a contagem de reticulócitos todos realizam a técnica manual com o reagente azul de cresil brilhante. Conclusão: O percentual de contagem de reticulócitos em relação aos hemogramas realizados em laboratórios do município de Divinópolis/MG é baixo, mesmo sendo o primeiro exame complementar para o diagnóstico diferencial da anemia. Essa baixa solicitação pode estar relacionada com a desvalorização do exame no meio clínico ou pela falta de acurácia e pobre reprodutibilidade dos resultados encontrados. (AU)

Objective: To estimate the percentage of reticulocyte counts in relation to blood count performed in the clinical analysis laboratories in Divinópolis / MG. Methods: This is a descriptive study carried out in the clinical analysis laboratories of Divinópolis/MG registered in the National Register of Health Establishment (CNES). A questionnaire interview was conducted in order to obtain information on the number of blood counts and reticulocyte counts performed during the last twelve months. The questionnaire also aimed to identify whether the institution was public or private, whether it was outpatient or hospital care and which methods were used to perform the tests. Results: Of the 15 laboratories registered in the CNES, 11 were interviewed (73%). 276,666 blood counts and 4,248 reticulocyte counts were performed in the last 12 months in all laboratories, which means the percentage of reticulocyte count of 1.5% in relation to the blood counts performed. Most of the laboratories are private (81%), medium-sized (54%) and carry out outpatient care (54%). To perform the blood count, most (81%) use the flow cytometry technique and for reticulocyte counting all perform the manual technique with the bright blue cresyl reagent. Conclusions: The percentage of reticulocyte counts in relation to blood counts performed in laboratories in the city of Divinópolis/MG is low, even though it is the first complementary exam for the differential diagnosis of anemia. This low request may be related to the devaluation of the reticulocyte in the clinical environment or the lack of accuracy and poor reproducibility of the results found. (AU)

Haematological values in a healthy adult population in Yaoundé, Cameroon

Background: Haematological values derived from local populations are useful in laboratories to improve diagnoses for local patients. In Cameroon, these data are not yet available. Moreover, there is great variation in baseline parameters pertaining to full blood cell count among medical laboratories.Objectives: This study aimed to determine values for the complete blood cell count of a healthy adult Cameroonian population for use in locally derived ranges in our medical laboratories.Methods: A cross-sectional study was conducted among blood donors attending three blood banks in Yaoundé from November 2015 to September 2016. We expected to obtain at least 120 venous blood samples from both men and women. Tests were performed for (1) HIV, (2) complete blood cell count, (3) hepatitis B virus, (4) malaria, (5) syphilis, (6) C-reactive protein and (7) hepatitis C virus.Results: We enrolled 294 healthy participants (161 men, 133 women) aged 18 to 55 years. The median haemoglobin concentration was 135 g/L in men and 114 g/L in women (p < 0.001). The median reticulocyte count was 60 × 109/L in men and 40 × 109/L in women (p < 0.001).Significant variation by sex was observed for the platelet count. The median white blood cell count was 4.1 × 109/L in men and 4.6 × 109/L in women (p = 0.008). Conclusion: This study provides locally derived ranges for complete blood cell and reticulocyte counts for a healthy adult population in Yaoundé, Cameroon. These results can be used pending larger studies

CRISPR/Cas9-mediated knockout of CD47 causes hemolytic anemia with splenomegaly in C57BL/6 mice / 한국실험동물학회지

CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein α (SIRPα) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout (CD47(−/−) hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, CD47(−/−) mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male CD47(−/−) mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female CD47(−/−) mice became evident at 26 weeks, but splenomegaly was identified in both genders of CD47(−/−) mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of CD47(−/−) mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our CD47(−/−) mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.

Cryohemolysis, erythrocyte osmotic fragility, and supplementary hematimetric indices in the diagnosis of hereditary spherocytosis

BACKGROUND: Hereditary spherocytosis (HS) is a chronic hemolytic anemia characterized by microspherocytes in the peripheral blood and increased erythrocyte osmotic fragility (EOF). This study evaluated the cryohemolysis test (CHT); initial hemolysis (IH); immediate and incubated hemolysis percentage in 5.5 g/L NaCl (H5.5); mean corpuscular hemoglobin concentration (MCHC); red blood cell distribution width (RDW); and Hb/MCHC, Hb/RDW, and MCHC/RDW ratios for the diagnosis of HS. METHODS: Data from 13 patients with HS were evaluated at the Instituto de Bioquímica Aplicada and compared with data from 14 unaffected individuals and 11 patients with anemia due to another etiology. Total blood and reticulocyte counts, CHT, and immediate and incubated EOF were performed in all subjects; sensitivity, specificity, efficiency, and Youden index (YI) were calculated. RESULTS: Eight patients with HS had MCHC ≥345 g/L, 10 had RDW ≥14.5%, 12 had IH >5.0 g/L, 11 had immediate H5.5 ≥5%, and 13 had incubated H5.5 ≥50% (the cut-off value to consider HS). The efficiency and YI were: immediate H5.5 (0.94–0.85), incubated H5.5 (0.89–0.82), IH (0.89–0.78), MCHC (0.87–0.62), CHT (0.84–0.54), and Hb/MCHC (0.71–0.56), respectively. The calculated ratios could distinguish subjects with HS from unaffected individuals (P 0.05). CONCLUSION: Although the CHT and supplementary hematimetric indexes were useful to differentiate individuals with SH from healthy controls, they cannot distinguish from anemias of other etiology. CHT and MCHC, in addition to EOF, are recommended for diagnosing HS patients because of their low cost and efficiency.

Neonatal indirect hyperbilirubinemia and glucose-6-phosphate dehydrogenase deficiency / 소아과

PURPOSE: This study aimed to determine the prevalence of glucose-6-phosphate dehydrogenase (G6PD) deficiency among infants with neonatal indirect hyperbilirubinemia (NIH); compare G6PD-deficient and G6PD-normal patients regarding hyperbilirubinemia and need for exchange transfusions (ET); and assess risk factors for ET and kernicterus. METHODS: This is a case-control retrospective study. Medical records of NIH patients admitted to the Pediatric Department, Salmaniya Medical Complex, Bahrain, between January 2007 and June 2010 were reviewed. Data on sex, age at presentation, hospitalization duration, need for ET, hemoglobin (Hb) level, reticulocyte count, direct Coombs test, serum total and indirect bilirubin levels, thyroid function, blood and urine cultures, G6PD status, and blood groups were collected and compared between the G6PD-deficent and G6PD-normal patients. RESULTS: Of 1,159 NIH patients admitted, 1,129 were included, of whom 646 (57%) were male. Among 1,046 patients tested, 442 (42%) were G6PD deficient, 49 (4%) needed ET, and 11 (1%) had suspected Kernicterus. The G6PD-deficient patients were mainly male (P<0.0001), and had lower Hb levels (P<0.0001) and higher maximum bilirubin levels (P=0.001). More G6PD-deficient patients needed ET (P<0.0001). G6PD deficiency (P=0.006), lower Hb level (P=0.002), lower hematocrit count (P=0.02), higher bilirubin level (P<0.0001), higher maximal bilirubin level (P<0.0001), and positive blood culture result (P<0.0001) were significant risk factors for ET. Maximal bilirubin level was a significant risk factor for kernicterus (P=0.021) and independently related to ET (P=0.03). CONCLUSION: G6PD deficiency is an important risk factor for severe NIH. In G6PD-deficent neonates, management of NIH should be hastened to avoid irreversible neurological complications.

Reticulocytosis in screen-printing workers exposed to 2-butoxyethanol and 2-ethoxyethanol

BACKGROUND: Studies on the hematologic toxicity of ethylene glycol ethers in humans are limited. Therefore, the aim of this study was to examine the association between exposure to solvents (containing 2-butoxyethanol and 2-ethoxyethanol) and hematological effects. METHODS: Thirty-four screen-printing workers who were exposed to 2-butoxyethanol and 2-ethoxyethanol and 37 non-exposed clerical workers were selected using data from the health care facilities that provided regular health screening services. Student's t-tests and Pearson's chi-square tests were used to compare differences in hematological parameters between the exposed and the control groups. A multivariate analysis was performed using the multiple logistic regression models to adjust for other variables. RESULTS: The chi-square test showed the reticulocyte percentages and corrected reticulocyte counts to be significantly higher in the exposed group. The t-tests showed a significant increase in white blood cell counts, reticulocyte percentages, and corrected reticulocyte count (i.e., reticulocyte index) in the exposed group, with p-values of 0.002, 0.004, and 0.002, respectively. Multivariate analysis showed the odds ratio for the corrected reticulocyte counts to be 16.30 for the exposed group, when compared with that of the control group. CONCLUSIONS: Exposure to 2-butoxyethanol and 2-ethoxyethanol was significantly associated with reticulocytosis, necessitating the implementation of preventive measures for workers prone to occupational exposure to ethylene glycol ethers.

Hb Heathrow β103(G5)Phe→Leu, a First Report in an Asian Patient with Erythrocytosis

Congenital erythrocytosis (CE) is a rare and heterogeneous disease. The high oxygen affinity hemoglobin (Hb) variants are the most common cause of CE. Herein, we report a Korean patient with isolated erythrocytosis. A 25-year-old man was referred to our hospital for evaluation of high Hb level (Hb 20.4 g/dL, hematocrit 58%, reticulocyte count 2.90%, white blood cell count 6.83×10⁹/L, and platelet count 195×10⁹/L). Bone marrow biopsy revealed normocellular marrow without myeloproliferative features. JAK2 (V617F, exon 12), CALR (exon 9), and MPL W515K/L mutations were not detected. P₅₀ (partial pressure at which Hb is half saturated with oxygen), which is an indicator of left-shift of oxygen dissociation curve (high oxygen affinity state), was 14.3 mm Hg (reference value 22.6–29.4 mm Hg). He was suspected to have CE. Mutation analysis of the HBB gene revealed the known Hb variant, Hb Heathrow [β103(G5)Phe→Leu]. This is the first report of Hb Heathrow in Asian.

Determination of reference ranges for immature platelet and reticulocyte fractions and reticulocyte hemoglobin equivalent

ABSTRACT Introduction: The immature platelet and immature reticulocyte fractions represent the ratios of platelets and reticulocytes recently released into the circulation and thus with higher RNA content. They are considered early indicators of bone marrow recovery. Objective: The aim of this study was to determine the reference ranges for the immature platelet and reticulocyte fractions of hematologically normal individuals in a university hospital. Methods: Venous blood samples collected in ethylenediaminetetraacetic acid K3 were analyzed using a Sysmex XE-5000™ analyzer. Individuals with platelet and reticulocyte counts within the reference ranges, and a blood count within the laboratory's screening criteria were included. Individuals with clinical conditions that could affect hematological results were excluded. The immature platelet fraction, high, medium and low fluorescence reticulocyte fractions and reticulocyte hemoglobin equivalent were evaluated. The reference ranges were determined according to the recommendations of the International Federation of Clinical Chemistry. Results: One hundred and thirty-two outpatients were evaluated. The mean age was 44 years (range: 13-80 years), 72 (54.5%) were women treated in a university hospital. The mean platelet count was 250.8 × 109/L and the mean reticulocyte count was 0.052 × 109/L. The following reference ranges were obtained: immature reticulocyte fraction 1.6-12.1%, the high, medium and low fluorescence reticulocyte fractions were 0.0-1.7%, 1.6-11.0% and 87.9-98.4%, respectively, the reticulocyte hemoglobin equivalent was 30.0-37.6% and immature platelet fraction was 0.8-5.6%. There was a statistically significant difference (p-value = 0.006) between genders in respect to the immature platelet fraction with 0.8-4.7% for females and 0.7-6.1% for males. The immature reticulocyte fraction was directly correlated with the reticulocyte count. Conclusion: Determining the reference range is critical to the introduction of a new parameter. The reference ranges obtained herein corroborate those reported in previous publications and will contribute to the clinical and laboratory application of the indices.

A Case of Myelodysplastic Syndrome Characterized by Hemolytic Anemia at Presentation

A man aged 78 yr with no history of chemotherapy or toxic exposure presented with a history of dyspnea and intermittent red urine for 3 months and several years, respectively. Hematologic data at admission were as follows: hemoglobin, 65 g/L; white blood cell count, 4.05x109/L; platelet count, 96x109/L; and reticulocyte count, 10.9%. A peripheral blood smear revealed polychromasia, nucleated red blood cells, and neutrophils with a non-lobulated nucleus. The bone marrow was hypercellular and exhibited an increase in erythroid precursors with trilineage dysplasia and our findings were suggestive of refractory cytopenia with multilineage dysplasia (RCMD). Karyotype of bone marrow cells was as follows: 45,XY,der(9;17)(p10;q10),add(18)(q11.2)[10]/45,idem,del(3)(q21)[10]. Other laboratory findings showed decreased serum haptoglobin, increased lactate dehydrogenase, and increased indirect bilirubin levels. Moreover, results of the direct/indirect antiglobulin test (Coombs' test) and paroxysmal nocturnal hemoglobinuria analysis with CD55, CD59, fluorescent aerolysin (FLAER), and CD24 were negative. Cold agglutinin and Donath-Landsteiner antibodies were not detected. This is a case of myelodysplastic syndrome (MDS) associated with hemolytic anemia and complex chromosomal abnormalities at presentation.

Effects of pre-immunosupressive therapy iron overload on hematologic response of severe aplastic anemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of serum ferritin (SF) and iron overload (IO) pre-immunosupressive treatment (IST) on hematologic response of severe aplastic anemia (SAA/VSAA) patients treated with IST.</p><p><b>METHODS</b>257 SAA/VSAA patients who underwent first-line IST from Feb, 2003 to Dec, 2011 in Anemia Therapeutic Centre, Institute of Hematology and Blood Diseases Hospital were retrospectively analyzed, the status of SF before IST and the IO-affected factors were studied. The effects of IO on hematologic response of SAA/VSAA patients were evaluated as well.</p><p><b>RESULTS</b>The median level of SF of 257 patients was 387 (6-2 004) μg/L. 36 patients (14%) had IO, including 20 SAA and 16 VSAA patients. According to univariate logistical regression analyses, IO was influenced by age>14 years (P=0.010) and blood transfusion (P<0.001). The multivariate logistic regression analysis showed that blood transfusion [P=0.001, OR=0.218 (95% CI 0.092-0.520)] was the only independent prognostic factor. SAA (but not for VSAA) patients with IO had much lower hematologic response rate in 6 month after IST (P=0.037). Absolute reticulocyte count and IO correlated with response at 6 month by univariate logistical regression analysis (P=0.014, 0.037). The multivariate logistic regression analysis showed that IO [P=0.021, OR=4.092 (95% CI 1.235-13.563)], ARC ≥20×10(9)/L [P=0.040, OR=2.743 (95% CI 1.049-7.175)] were independent prognostic factors.</p><p><b>CONCLUSION</b>84.8% patients had high serum ferritin before IST, and 14.0% reached IO. Adult and more blood transfusion caused IO more likely. IO correlated with response at 6 month, and was independent prognostic factor.</p>

A 2-month-old boy with hemolytic anemia and reticulocytopenia following intravenous immunoglobulin therapy for Kawasaki disease: a case report and literature review / 소아과

Herein, we report a rare case of hemolytic anemia with reticulocytopenia following intravenous immunoglobulin therapy in a young infant treated for Kawasaki disease. A 2-month-old boy presented with fever lasting 3 days, conjunctival injection, strawberry tongue, erythematous edema of the hands, and macular rash, symptoms and signs suggestive of incomplete Kawasaki disease. His fever resolved 8 days after treatment with aspirin and high dose infusion of intravenous immunoglobulin. The hemoglobin and hematocrit decreased from 9.7 g/dL and 27.1% to 7.4 g/dL and 21.3%, respectively. The patient had normocytic hypochromic anemia with anisocytosis, poikilocytosis, immature neutrophils, and nucleated red blood cells. The direct antiglobulin test result was positive, and the reticulocyte count was 1.39%. The patient had an uneventful recovery. However, reticulocytopenia persisted 1 month after discharge.

Performance Evaluation of the DxH 800 Hematology Analyzer

In this study, the performance of a hematology analyzer, DxH 800 (Beckman Coulter, USA) was evaluated. The precision, carry-over, linearity, and comparison studies were performed according to the Clinical Laboratory Standards Institute guidelines. The test items were white blood cell, red blood cell, hemoglobin, red blood cell index, platelet, and reticulocyte counts. The 6C control and Retic-X cell control (Beckman Coulter) were used for precision evaluation. For the correlation study, the test results were compared with those obtained from the ADVIA 2120i (Siemens, USA) using 120 blood samples. The results of precision and carry-over evaluations were within acceptable range. The coefficient of determination (R 2) for linearity was good, being more than 0.99. The correlation coefficient (R) ranged from 0.945 to 0.996. DxH 800 was evaluated as an acceptable hematology analyzer for the automation of large volume of laboratory samples.

El conteo automático de reticulocitos: una herramienta de uso diagnóstico, clínico e investigativo / Automated reticulocyte count: a tool for diagnostic, clinical and research use

Se revisan los antecedentes históricos del conteo de reticulocitos como determinación indispensable en el laboratorio de hematología para la evaluación de la actividad eritropoyética durante la clasificación, diagnóstico y monitoreo de la respuesta terapéutica en distintos trastornos y situaciones clínicas, principalmente en casos de anemias. Se describe el tránsito del tradicional método de conteo de reticulocitos manual al método automatizado y la integración de los parámetros reticulocitarios al hemograma automatizado actual; además, se analizan las desventajas del método de recuento manual y las ventajas del método automatizado, así como los principios de detección en que se basa el conteo electrónico de reticulocitos. Con relación a los parámetros reticulocitarios, se describe su medición, cálculo y unidades de medida; también se resalta la importancia de la fracción de reticulocitos inmaduros y del contenido de hemoglobina reticulocitaria como variables de mayor uso clínico e investigativo en la evaluación de la respuesta medular ante diversos trastornos clínicos y protocolos terapéuticos. Por último, se alude a la necesidad del conocimiento y empleo de las variables reticulocitarias en la práctica clínica de rutina por parte de los clínicos y especialistas en hematología.

The historical background of reticulocyte count is reviewed as an essential determination in the laboratory of hematology for the evaluation of erythropoietic activity during classification, diagnosis and monitoring of therapeutic response of different conditions and clinical situations are also reviewed, especially in anemia The transition from traditional manual reticulocyte counting method to automated method and integration of the reticulocyte parameters to current automated complete blood count are described. The disadvantages of manual method and the advantages of automated methods are cited, as well as detection principles in which electronic reticulocyte count is based. Regarding reticulocyte parameters, measurement, calculation and units are described. The importance of immature reticulocyte fraction and reticulocyte hemoglobin content as variables most clinical and research use in evaluating bone marrow response to various clinical disorders and therapeutic protocols are highlighted. Finally, the need for knowledge and use of reticulocyte variables in routine clinical practice by clinicians and hematologist is referred.

Value of MCV/RDW Combined with Reticulocyte Parameters in Differential Diagnosis of Anemia Diseases / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the value of mean corpuscular volume/RBC distribution width (MCV/RDW) combined with reticulocyte parameters in differential diagnosis of aplastic anemia (AA), myelodysplastic syndrome (MDS), megaloblastic anemia (MA) and hemolytic anemia (HA) in order to provide some laboratorial evidence for clinical doctors in first diagnosis of these diseases.</p><p><b>METHODS</b>The data of MCV/RDW and reticulocyte parameters of AA, MDS, MA and HA patients from January 1 of 2011 to August 31 of 2014 were retrospectively collected in West China Hospital of Sichuan University. And 158 healthy unrelated individuals with age-, sex-matched were collected as controls. The value of MCV/RDW and reticulocyte parameters in differentiating diagnosis of above mentioned 4 kinds of anemia diseases was assessed. ROC analysis was used to determine the cutoff value of MCV/RDW and the reticulocyte parameters were performed in differentiating diagnosis of AA and MDS.</p><p><b>RESULTS</b>The average values of MCV/RDW of 158 AA patients (79 acute AA patients and 79 chronic AA patients), 107 MDS patients, 13 MA patients and 81 HA patients increased in variable degrees as compared with the controls, and there was statistical difference between them, the MCV/RDW value of acute AA patients was obviously less than that of other patients. In the 4 kinds of anemia diseases, the reticulocyte absolute count in acute AA patients was the lowest, that of chronic AA, MA and MDS patients was higher, and that of HA patients was highest. The ratio of low fluorescent reticulocyte decreased, and the ratio of moderate and high fluorescent reticulocytes increased in the 4 kinds of anemia diseases, as compared to controls. The difference was statistically significant. The analysis of differential diagnosis of chronic AA and MDS showed that RDW-SD could differentiate the chronic AA from MDS. The area under the curve (AUC) of RDW-SD was 0.76 (P < 0.01). The cutoff value of RDW-SD was 22.75fl. The sensitivity and specificity of RDW-SD for differential of chronic AA and MDS was 49.5% and 98.7%, respectively.</p><p><b>CONCLUSION</b>MCV/RDW and reticulocyte parameters can be used as the laboratorial differential diagnostic indicators for AA, MDS, MA and HA diseases.</p>

A 14-Day Repeated Dose Toxicity of Epimedii Herba Aqueous Extract Administered by Oral Gavage in F344 Rats

The objective of this study is to characterize a toxicity of Epimedii Herba (EH) in F344 rats and to find a dose levels for the 13 weeks toxicity study. EH is well known as medicinal herb in many Asian countries for traditional medicines of antibacterial and antiviral effects, estrogenic and antiestrogenic effects, and for treatment of osteoporosis, hypotensives, fatigue, kidney disorders, and related complications. However, the indispensable and basic information of toxicological evaluation of EH extract is insufficient to support its safe use. Therefore, we conducted toxicological evaluation of this drug in compliance with OECD and MFDS guideline in this study. The extract of EH was administered orally to F344 rats at dose levels of 0, 500, 1000, 2000, 3500, and 5000 mg/kg/day for 2 weeks. Each group was composed of 5 male and female rats. In this study, there were no treatment of EH-related adverse changes in clinical observations, mortality, body weights, food consumption, urinalysis, gross finding at necropsy, and organ weight examination. Total red blood cell count, hematocrit, mean corpuscular hemoglobin concentration, total cholesterol, and phospholipid were decreased in males and females at 5000 mg/kg/day compared to the control animals. Mean corpuscular volume and reticulocyte counts were increased in males and females at 5000 mg/kg/day compared to control animals. Therefore, we recommend that dose level of 5000 mg/kg/day is a highest treatment group in 13-week EH extract exposure study for further toxicity assessment.

A Comparison of the Performance of Soyagreentec Ampulab EDTA and Sodium Citrate Tubes with That of BD Vacutainer Tubes

BACKGROUND: Because blood collection tube can affect the results of various laboratory tests, it is necessary before using a newly developed product to verify its performance stringently and objectively. We compared the performance of Ampulab EDTA and sodium citrate tubes (Soyagreentec, Korea) with that of Vacutainer tubes (BD, USA) in accordance with international guidelines. METHODS: This study was performed as a multicenter study of Chung-Ang University Hospital and Seoul National University Hospital to evaluate the performance of two different instrument platforms. We performed the precision test according to CLSI GP34-A, the accuracy test according to CLSI EP09-A2-IR, and the vacuum test according to CLSI H1-A5 as well as stability, and aseptic condition tests. We evaluated 3 lots of Ampulab tubes for their precision, accuracy, vacuum, and aseptic condition. RESULTS: In precision test, the total precision levels of Ampulab tubes in most measurands were desirable or allowable. The results of mean corpuscular hemoglobin concentration, platelet distribution width, basophil, and reticulocyte counts for Ampulab tubes showed imprecision beyond allowable limits, but were similar to those of Vacutainer tubes. In the accuracy test, the bias in most measurands, except for the mean platelet volume, was within allowable limits. In the stability test, Ampulab showed similar performance to Vacutainer. In tests of the vacuum and aseptic conditions, Ampulab fulfilled both requirements. CONCLUSIONS: The performance of Ampulab EDTA and sodium citrate tubes was equivalent to that of Vacutainer tubes.