Generation of hematopoietic stem/progenitor cells with sickle cell mutation from induced pluripotent stem cell in serum-free system

ABSTRACT Introduction Sickle cell disease (SCD) is a monogenic disease and it is estimated that 300,000 infants are born annually with it. Most treatments available are only palliative, whereas the allogeneic hematopoietic stem cell transplantation offers the only potential cure for SCD. Objective Generation of human autologous cells, when coupled with induced pluripotent stem cell (iPSC) technology, is a promising approach for developing study models. In this study, we provide a simple and efficient model for generating hematopoietic cells using iPSCs derived from a sickle cell anemia patient and an inexpensive in-house-prepared medium. Method This study used iPSCs previously generated from peripheral blood mononuclear cells (PBMCs) from a patient with sickle cell anemia (iPSC_scd). Hematopoietic and erythroid differentiation was performed in two steps. Firstly, with the induction of hematopoietic differentiation through embryoid body formation, we evaluated the efficiency of two serum-free media; and secondly, the induction of hematopoietic stem/progenitor cells to erythroid progenitor cells was performed. Results The patient-specific cell line generated CD34+/CD45+ and CD45+/CD43+ hematopoietic stem/progenitor cells and erythroid progenitors, comprising CD36+, CD71+ and CD235a+ populations, as well as the formation of hematopoietic colonies, including erythroid colonies, in culture in a semi-solid medium. Conclusion In conjunction, our results described a simple serum-free platform to differentiate human the iPSCs into hematopoietic progenitor cells. This platform is an emerging application of iPSCs in vitro disease modeling, which can significantly improve the search for new pharmacological drugs for sickle cell disease.

Comparison of Effect of Serum-Free Culture Systems on Directional Erythroid Differentiation of Human Umbilical Cord Blood CD34 Cells / 中国实验血液学杂志

OBJECTIVE@#To compare the efficacy of directional erythroid differentiation in different serum free culture systems and to screen the optimal culture systems for inducing the differentiation of umbilical cord blood hematopoietic stem and progenior cells (HSPC) to erythroid cells.@*METHODS@#The CD34 cells from umbilical blood munonuclear cells were sorted by using the magnetic beads, and were inoculated into 3 different of culture systems (system 1, 2 and 3 respectively), to induce erythrold differentiation by 3 stage culture. The living cells were counted in different differentiation stages and were observed by Wright-Giemsa staining; the expression of CD71 and CD235a on cell surface was detected by flow cytometry, the erythroid differentiation pteency was detected via colony-forming test.@*RESULTS@#The ability of system 2 to promote the HSPC proliferation was the strongest, the efficacy of system 3 to promote the erythroid differentiation of HSPC was the most optimal; the proliferation ability of cells cultured in system 2 for 2-15 days all was higher than that of cells cutured in system 1 and 3 (P＜0.05). The flow cytometry detection showed that the expression of CD71 and CD235a on surface of cells cultured in system 3 was the highest, the CD235a percentage on day 15 of differentiation in system 3 was (92.33±3.89)%, that in system 2 was (84.67±3.12)%, while that in system 1 was (72.17±6.83)% (P＜0.05). Cell morplologic detection showed that throid differentiation was accelerated on day 12, the percentage of orthochromatic erythrocytes in system 3 was (67.67±2.08)% which was 10.69 and 25.34 times higher than that in system 2 and 1 respectively (P＜0.05). The colony-forming test showed the ratio of BFU-E in system 3 increased gradually on day 3-9 (r=0.99, P＜0.05), which was significanlly higher than that in system 2 and 1 on day 9 (90.35±5.52% vs 77.06±2.26% and 74.50±3.95%).@*CONCLUSION@#Culture system 3 is the most effective serum-free erythroid differentiation system, and the culture system 2 is the most powerful HSPC proliferation system. This study results provide a technical basis for further efficiently increasing and inducing the erythroid proliferation and differentiation of HSPC, and also provide culture system in vitro for the clinical application and basic research.

Anemia de Diamond Blackfan: un diagnóstico de exclusión / Anemia of Diamond Blackfan: an exclusión diagnosis

La anemia de Diamond Blackfan es un trastorno genético y clínico raro, caracterizado por aplasia eritrocitaria, que clásicamente se manifiesta durante el primer año de vida, típicamente a los 2-3 meses de edad. El 25% de los afectados presentan anemia severa en la infancia, normo o macrocitosis, reticulocitopenia y disminución selectiva de células precursoras eritroides en medula ósea. Es causada por mutaciones que afectan genes que codifican para proteínas ribosomales, inicialmente fue identificado RPS19, que codifica la proteína S19 y las mutaciones a otros genes que codifican proteínas ribosomales. Se presenta el caso de una paciente de cuatro meses de edad quien debutó con anemia severa, quien no mejoró con la suplencia de hierro, vitamina B12, y ácido fólico y además fueron descartadas sistemáticamente causas frecuentes de anemia. El diagnóstico de anemia de Diamond Blackfan en nuestro medio es un diagnóstico de exclusión, dada la dificultad para acceso a pruebas de confirmación genética. Se establece el diagnóstico y se da manejo con glucocorticoides con buena respuesta clínica y paraclínica

The Diamond Blackfan anemia is a rare genetic and clinical disorder. It is characterized by red cell aplasia, which typically occurs during the first year of life, typically during the second to the third month of age. 25% of the patients had severe anemia during their childhood, normo or macrocytosis, reticulocyte and selective decrease in the number of erythroid precursor cells in bone marrow. It is caused by mutations affect genes encoding ribosomal proteins, RPS19 initially was identified, which encodes S19 protein and mutations in other genes encoding ribosomal proteins. We present a case of a four-month-old who debuted with severe anemia in whom the substitution were iron supplements, vitamin B12 and folic acid, showed no improvement and who also were systematically discarded as common causes of anemia. The diagnosis of Diamond Blackfan anemia in our country is a diagnosis of exclusion, given the difficulty of access to genetic confirmation tests. In this article the diagnosis is established and gives management with glucocorticoid with good clinical and paraclinical response

Human parvovirus B19 in patients with beta thalassemia major from Tehran, Iran

BACKGROUND: Due to the tropism of human parvovirus B19 to erythroid progenitor cells, infection in patients with an underlying hemolytic disorder such as beta-thalassemia major leads to suppression of erythrocyte formation, referred to as transient aplasia crisis (TAC), which may be life-threatening. We investigated the prevalence of parvovirus B19 among patients with beta thalassemia major attending the Zafar Adult Thalassemia Clinic in Tehran, Iran. METHODS: This cross-sectional study was performed to determine the presence of parvovirus B19 DNA in blood samples and parvovirus B19 genotypes in plasma samples of patients with thalassemia major. The population consisted of 150 patients with beta-thalassemia major who attended the Zafar clinic in Tehran. Specimens were studied using a real-time polymerase chain reaction assay. RESULTS: The prevalence of parvovirus B19 in our study population was 4%. Of 150 patients with thalassemia, six (4%) were positive for B19 DNA. There was no significant correlation between blood transfusion frequency and B19 DNA positivity. Finally, phylogenetic analysis of human parvovirus B19 revealed genotype I in these six patients. CONCLUSION: In this study, acute B19 infections were detected in patients with beta thalassemia major. Screening of such high-risk groups can considerably reduce the incidence and prevalence of B19 infection; thus, screening is required for epidemiologic surveillance and disease-prevention measures.

Significance of Morphological Examination, Cytochemical Staining Combined with Bone Marrow Biopsy in Differential Diagnosis of Myelodysplastic Syndrome with Low Blasts and Hemolytic Anemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the value of morphological examination, cytochemical staining combined with bone marrow biopsy in the differential diagnosis between myelodysplastic syndrome (MDS) with low blasts and hemolytic anemia (HA).</p><p><b>METHODS</b>The clinical data of 85 cases of myelodysplastic syndrome with low blasts (< 5%) and 61 patients with hemolytic anemia in Chinese PLA's Gerneral hospital from September 2009 to March 2015 were retrospectively analysed. The clinical characteristics, cytogenetic and molecular features, bone marrow cell count and morphology features, cytochemical staining results and bone marrow biopsy features of above-methioned patients were compared.</p><p><b>RESULTS</b>There was no significant difference (P > 0.05) in clinical data between MDS group and HA group. Megakaryocytic dysplasia-positive rate, and ring sideroblasts positive rate, and PAS positive rate were significantly higher in MDS group than those that in HA group (P < 0.05). Abnormal localization of immature precursors (ALIP) and megakaryocytic dysplasia positive rate in bone marrow biopsy were significantly higher in MDS group than those that in HA group (P < 0.05), 90.6% of MDS with low blasts patients were identifiable by combined detections.</p><p><b>CONCLUSION</b>Combining detection of morphology, cytochemistry staining and bone marrow biopsy has been confirmed to be more useful for differential diagnosis between MDS with low blasts and HA.</p>

Relationship among the Oxygen Concentration, Reactive Oxygen Species and the Biological Characteristics of Mouse Bone Marrow Hematopoietic Stem Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of oxygen concentration and reactive oxygen species (ROS) on the biological characteristics of hematopoietic stem cells (HSC) and to analyzed the relationship among the oxygen concentration, ROS and the biological characteristics of mouse HSC through simulation of oxygen environment experienced by PB HSC during transplantation.</p><p><b>METHODS</b>The detection of reactive oxygen species (ROS), in vitro amplification, directional differentiation (BFU-E, CFU-GM, CFU-Mix), homing of adhesion molecules (CXCR4, CD44, VLA4, VLA5, P-selectin), migration rate, CFU-S of NOD/SCID mice irradiated with sublethal dose were performed to study the effect of oxgen concentration and reactive oxygen species on the biological characteristics of mouse BM-HSC and the relationship among them.</p><p><b>RESULTS</b>The oxygen concentrations lower than normal oxygen concentration (especially hypoxic oxygen environment) could reduce ROS level and amplify more Lin(-) c-kit(+) Sca-1(+) BM HSC, which was more helpful to the growth of various colonies (BFU-E, CFU-GM, CFU-Mix) and to maintain the migratory ability of HSC, thus promoting CFU-S growth significantly after the transplantation of HSC in NOD/SCID mice irradiated by a sublethal dose. BM HSC exposed to oxygen environments of normal, inconstant oxygen level and strenuously thanging of oxygen concentration could result in higher level of ROS, at the same time, the above-mentioned features and functional indicators were relatively lower.</p><p><b>CONCLUSION</b>The ROS levels of BM HSC in PB HSCT are closely related to the concentrations and stability of oxygen surrounding the cells. High oxygen concentration results in an high level of ROS, which is not helpful to maintain the biological characteristics of BM HSC. Before transplantation and in vitro amplification, the application of antioxidancs and constant oxygen level environments may be beneficial for transplantation of BMMSC.</p>

The induction and cryopreservation of erythroid progenitor cells derived from umbilical cord blood mononuclear cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To discover the techniques for ex vivo generation and cryopreservation of erythroid progenitor cells (EPCs)derived from umbilical cord blood (UCB)mononuclear cells (MNCs).</p><p><b>METHODS</b>UCB was chosen as the source of EPCs. Erythrocytes were precipitated by hydroxyethyl starch (HES). MNCs were separated by Ficoll density gradient centrifugation. Erythroid progenitor cell were generated from MNC ex vivo in suspension culture supplemented with stem cell growth factor, insulin growth factor, erythropoietin, Fms- liketyrosinekinase ligand, transferrin and dexamethasone. Cell maturation was evaluated by morphologic analysis and CD71/CD235a expression profiling. In vitro induced cells were cryopreserved using different cryopreservation media. The cell survival rate, phenotype and proliferation curves were detected after cell thawing.</p><p><b>RESULTS</b>With the extension of culture time, the total number of cells increased significantly accompanied with the elevation of CD71 and CD235 positive populations. After 14- day inducing, the cells reached to approximately 110 times of the starting number with the cell viability as (88.92±0.95)%. The percentages of cell surface markers were (86.77±9.11)% for CD71 and (64.47±16.67)% for CD71/CD235, respectively. With the extension of inducing time, wright- Giemsa staining showed that the middle erythroblasts appeared mostly at day 10, and the late erythroblasts were seen at day 14. The red pellets were present at day 14, which indicated the more production of hemoglobin. Colony forming assay showed that erythroid colonies at induction day 7 were higher than that for non-induced cells (326.00±97.96vs 61.60±20.03 per 2 000 cells). With the extension of culture time, the number of erythroid colonies decreased. Induced EPCs were preserved with different cryopreservation solutions, in which 10% DMSO were better than 5% DMSO. Additionally, 10% DMSO + 2% HSA showed no different with 10% DMSO + 5% HSA. Combined 50% plasma with 2% HSA was more effective.</p><p><b>CONCLUSIONS</b>This non- serum culture media could effectively induced and expanded EPCs, and 10% DMSO + 2% HSA + 50% plasma appeared to be a desirable cryopreservation solution for EPCs from UCB.</p>

As infecções genitais podem alterar os resultados dos testes preditivos do parto prematuro? / Can genital infections alter the results of preterm birth predictive tests?

OBJETIVOS: Verificar se a presença de agentes infecciosos no conteúdo vaginal ou cervical pode alterar os resultados dos testes da proteína-1 fosforilada ligada ao fator de crescimento insulina-símile (phIGFBP-1) e das medidas do comprimento do colo uterino (CC) pela ultrassonografia transvaginal. MÉTODOS: Um total de 107 gestantes com antecedente de prematuridade espontânea foram submetidas ao teste da phIGFBP-1 e à realização da ultrassonografia transvaginal para medida do comprimento do colo uterino, a cada três semanas, entre 24 e 34 semanas. As infecções genitais foram pesquisadas imediatamente antes da realização dos testes. As pacientes foram distribuídas em quatro grupos (GA, GB, GC e GD) e dentro de cada grupo foi avaliada a correlação entre infecção genital e alteração nos testes utilizando a análise das razões de chance (OR) e o coeficiente de correlação de Pearson. RESULTADOS: Em cada grupo, mais de 50% das pacientes apresentaram infecção genital (GA 10/17; GB 28/42; GC 15/24; GD 35/53), sendo a vaginose bacteriana a principal alteração de flora vaginal. O resultado positivo para phIGFBP-1 (GA 10/10; GB 18/28; GC 15/15; GD 19/35) e CC≤20 mm (GA 10/10; GB 20/28; GC 10/15; GD 20/35) foram os resultados encontrados com maior frequência nas pacientes com infecção genital em todos os grupos. Porém, aplicando o coeficiente de correlação de Pearson foi identificada correlação entre infecção genital e positividade para os marcadores. CONCLUSÃO: A presença de alteração da flora vaginal e de outras infecções genitais não alteram significativamente os resultados do teste da phIGFBP-1 e da medida do colo uterino quando comparados aos casos sem infecção. No entanto, é necessária ...

PURPOSE: To determine if the presence of infectious agents in vaginal or cervical content can alter the results of the insulin-like growth factor binding protein-1 (phIGFBP-1) test and the measurement of cervical length (CC) by transvaginal ultrasonography. METHODS: A total of 107 pregnant women with a history of spontaneous preterm birth were submitted to the phIGFBP-1 test and to measurement of CC by transvaginal ultrasonography every 3 weeks, between 24 and 34 weeks of gestation. Genital infections were determined immediately before testing. The patients were distributed into four groups (GA, GB, GC, and GD) and the correlation between genital infection and changes in the tests was determined within each group based on the odds ratio (OR) and the Pearson correlation coefficient. RESULTS: In each group, over 50% of the patients had genital infections (GA 10/17; GB 28/42; GC 15/24; GD 35/53), with bacterial vaginosis being the main alteration of the vaginal flora. Positive results for phIGFBP-1(GA 10/10; GB 18/28; GC 15/15; GD 19/35) and CC≤20 mm (GA 10/10; GB 20/28; GC 10/15; GD 20/35) were obtained more frequently in patients with genital infection in all groups. Nonetheless, when applying the Pearson correlation coefficient we detected a poor correlation between genital infection and positivity for markers. CONCLUSION: The presence of changes in the vaginal flora and of other genital infections does not significantly alter the results of phIGFBP-1 and the measurement of cervical length when compared to cases without infection. However, more studies with larger samples are necessary to confirm these results. .

Comparison of in-vitro and in-vivo response to fetal hemoglobin production and γ-mRNA expression by hydroxyurea in Hemoglobinopathies.

BACKGROUND: Hydroxyurea, which induces Fetal hemoglobin (HbF) synthesis, is the only drug widely used in different hemoglobinopathies; however, the response is very variable. We compared the efficacy of hydroxyurea in-vitro in erythroid cultures and in-vivo in the same patients with different hemoglobinopathies to induce HbF production and enhance γ-messenger RNA expression. MATERIALS AND METHODS: A total of 24-patients with different Hemoglobinopathies were given hydroxyurea and their response was studied in-vivo and in-vitro on mononuclear cells collected from them simultaneously. RESULTS: A total of 57.7% of patients (responders) showed no further crisis or transfusion requirements after hydroxyurea therapy with a mean increase in fetal cells (F-cells) of 63.8 ± 59.1% and γ-mRNA expression of 205.5 ± 120.8%. In-vitro results also showed a mean increase in F-cells of 27.2 ± 24.7% and γ-mRNA expression of 119.6% ± 65.4% among the treated cells. Nearly 19.0% of the partial-responders reduced their transfusion requirements by 50% with a mean increase in F-cells of 61.2 ± 25.0% and 28.4 ± 25.3% and γ-mRNA-expression of 21.0% ± 1.4% and 80.0% ± 14.1% in-vivo and in-vitro respectively. The non-responders (15.3%) showed no change in their clinical status and there was no significant increase in F-cells levels and γ-mRNA expression in-vivo or in-vitro. CONCLUSION: Thus, this method may help to predict the in-vivo response to hydroxyurea therapy; however, a much larger study is required.

Diagnostic value of dysplasia characteristics in typing of myelodysplastic syndrome / 中国实验血液学杂志

This study was purpose to investigate the diagnostic value of hematopoietic cell dysplasia in myelodysplastic syndrome (MDS). Sixty-four cases of WHO-MDS were detected by cytomorphology, cytohistochemical staining and bone marrow biopsy. The characteristics of hematopoietic cell dysplasia were analyzed, and its sensitivity and specificity were evaluated for WHO-MDS diagnosis. The results showed that though myeloblast, megakaryocytes presented in peripheral blood and granular Auer body, abnormal granular pseudo Pelger-Huër, vacuole of erythroid, micro-megakaryocytes appeared in bone marrow for diagnosis sensitivity were not very high, and respectively were 34.4%, 3.1%, 3.1%, 75.0%, 6.3%, 42.4%, the specificity of these characteristics was 100%. Moreover, erythroid odd nucleus, nuclear deformity, fragmentation, nuclear budding, ring sideroblasts, single and more round nuclear megakaryocyte had better reference value for WHO-MDS diagnosis. By bone marrow biopsy, the dysplasia and abnormal localization of immature precursor (ALIP) also were found in patients with WHO-MDS. More than half patients with WHO-MDS had mild to moderate increase in reticulin fibres. It is concluded that the cytomorphology assay is the base and key for the diagnosis of WHO-MDS. Diagnostic accuracy can be improved by combinative use of a variety of detection methods.

Parvovirus B19 Infection in a Child with Acute Lymphoblastic Leukemia during Maintenance Chemotherapy / 임상소아혈액종양

Parvovirus B19 targets human erythroid progenitor cells, causing a self-limiting subclinical erythroid aplasia in the healthy hosts, whereas the immunocompromised subjects generate a prolonged viremia and chronic anemia with or without thrombocytopenia or neutropenia. The attenuated immune response in patients with acute lymphoblastic leukemia (ALL), receiving chemotherapy, may generate the hematologic aberration which mimic a leukemia relapse or therapy-induced cytopenia. This mimicry may lead to the unnecessary examination and the recess of chemotherapy. If the anemia with or without thrombocytopenia or neutropenia is noticed during the chemotherapy of ALL, the parvovirus B19 infection should be considered as a cause of hematologic aberration. We report a case of parvovirus B19 infection confirmed by PCR in a child with ALL who was initially sero-negative (IgM and IgG) against parvovirus B19 to highlight the importance of alertness to the possibility of parvovirus B19 infection during chemotherapy.

Effects of 1,4-benzoquinone on the proliferation activity of human bone marrow stem cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the influence of 1,4-benzoquinone (1,4-BQ) on proliferation of human bone marrow haematopoietic stem cells (hBM-HSCs) and human bone marrow mesenchymal stem cells (hBM-MSCs).</p><p><b>METHODS</b>The bone marrow samples were collected from a healthy donor. Methylcellulose semi-solid culture medium was used to culture the mononuclear cells of bone marrow in different culture systems. Colony-forming unit (CFU) assay was utilized to evaluate the proliferation of hBM-HSCs exposed to 1,4-BQ at the doses of 10, 25, 50 and 100 µmol/L and to observe the influence of 1,4-BQ on the Colony-forming unit-erythroid (CFU-E)/Burst-forming unit-erythroid (BFU-E), Colony-forming unit-granulocyte, macrophage (CFU-GM), Colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM) in hBM-MSCs. MTT assay was used to detect the proliferation of hBM-MSCs exposed to 1,4-BQ at the doses of 1, 5, 10, 25, 50, 100, 200, 500 and 1000 µmol/L for 24 h, respectively, after hBM-MSCs were isolated, cultured and expanded.</p><p><b>RESULTS</b>The results of CFU assay indicated that numbers of CFU-E/BFU-E, CFU-GM and CFU-GEMM in 25, 50 and 100 µmol/L groups significantly decreased, as compared with control group (P < 0.05). However, no significant difference was found between the 10 µmol/L group and the control group. The results of MTT assay showed that the cellular viability of hBM-MSCs exposed to 1,4-BQ at the doses of 50 ∼ 200 µmol/L for 24 h significantly decreased in a dose-depended manner. When the exposure dose was higher than 200 µmol/L, the cellular viability of hBM-MSCs was lower than 5% which was significantly lower than that of control group (P < 0.05). When the exposure dose was lower than 25 µmol/L, there was no significant difference of cellular viability between exposure group and control group (P > 0.05).</p><p><b>CONCLUSION</b>The results of the present study demonstrated that 1,4-BQ could inhibit the colony forming of hBM-HSCs and the relative viability of hBM-MSCs in vitro. The hematotoxicity induced by 1,4-BQ may be related to inhibiting the proliferation capacity of hBM-HSCs.</p>

Parvovirus B19 Infection in a Child with Acute Lymphoblastic Leukemia during Maintenance Chemotherapy / 임상소아혈액종양

Parvovirus B19 targets human erythroid progenitor cells, causing a self-limiting subclinical erythroid aplasia in the healthy hosts, whereas the immunocompromised subjects generate a prolonged viremia and chronic anemia with or without thrombocytopenia or neutropenia. The attenuated immune response in patients with acute lymphoblastic leukemia (ALL), receiving chemotherapy, may generate the hematologic aberration which mimic a leukemia relapse or therapy-induced cytopenia. This mimicry may lead to the unnecessary examination and the recess of chemotherapy. If the anemia with or without thrombocytopenia or neutropenia is noticed during the chemotherapy of ALL, the parvovirus B19 infection should be considered as a cause of hematologic aberration. We report a case of parvovirus B19 infection confirmed by PCR in a child with ALL who was initially sero-negative (IgM and IgG) against parvovirus B19 to highlight the importance of alertness to the possibility of parvovirus B19 infection during chemotherapy.

Analysis of clinical characteristics in 45 cases of Diamond-Blackfan anemia / 中国实验血液学杂志

In order to explore the diagnosis and therapy of Diamond Blackfan anemia (DBA), the clinical data of 45 cases of DBA admitted in our hospital from February 1994 to July 2011 were analyzed retrospectively. The clinical characteristics, results of laboratory examination, treatment reaction and outcome of disease were investigated. The results indicated that out of 45 children diagnosed as DBA, 14 cases (31.1%) had short stature and physical malformation. All patients had anemia with reticulocytopenia. Thirty-four patients (75.6%) had mean corpuscular volume. Eleven patients (24.4%) had macrocytic anemia. Bone marrow examination showed a marked erythroid hypoplasia in all patients. Out of 29 cases tested for fetal hemoglobin (HbF), 13 cases (44.8%) had high level of HbF. Erythroid colony-forming unit of bone marrow was tested in 25 patients, among them 12 patients (48%) showed normal plasia, 13 (52%) showed hypoplasia. The erythropoietin (EPO) levels of 17 patients were elevated. Karyotypes were examined in 28 patients, and showed all normal. The treatment was based on corticosteroids and Cyclosporine A. Thirty patients had good response to corticosteroid therapy, and 10 of them obtained a sustained corticosteroid-induced remission. Twenty cases discontinued corticosteroid therapy after remission, as a result, 15 cases (75%) relapsed, moreover all the relapsed cases still had good response to corticosteroid. Two relapsed patients suffered from aplastic anemia, one of them died of therapy failure. Six patients were unresponsive to corticosteroid, 1 of which achieved remission with cyclosporine A and the others continued to receive regular transfusions. 3 patients received iron chelation therapy. It is concluded that the clinical characteristics, complete blood count, bone marrow smear, HbF level and EPO level are useful to make a diagnosis of DBA. Most patients have a good response to corticosteroid therapy, but relapse rate is high when drug was discontinued. Patients unresponsive to corticosteroid should receive regular transfusions and chelation therapy.

Interpretation of maturing trilineage hematopoiesis

Normal adult bone marrow occupies the medullary spaces of large bones. The main function of bone marrow is the production of blood cells [hematopoiesis]. Bone marrow is composed of a matrix requisite for hematopoiesis as well as erythriod precursors, myeloid precursors with monocyte macropliage system, megakaryocytes, lymphocytes, plasma cells, blood vessels and stroma. Interpretation of bone marrow maturing trilineage hematopoiesis includes the assessment of the maturation seq uence and morphologic features for each lineage. This review presents detailed guidelines for interpretation of bone marrow maturing trilineage hematopoiesis according to the cell line and the pathologic condition

TfR2 mRNA expression in bone marrow mononuclear cells of children with hyperplastic anemia and its implications / 中国实验血液学杂志

The aim of this study was to investigate the expression of transferrin receptor 2 (TfR2) mRNA in bone marrow mononuclear cells (BMMNC) of children with hyperplastic anemia (HA), to analyze the correlation of TfR2 mRNA expression level with Hb level, bone marrow erythroid hyperplasia, iron status in body and underlying diseases, and to evaluate the role of TfR2 in erythroid hemopoiesis and the useful value in diagnosis of HA. The experiment was divided into 2 groups: test group, in which 40 patients with HA were enrolled, and control group in which 10 patients without erythroid disorders and hematological malignancies confirmed by bone marrow examination were enrolled. The bone marrow samples of patients in mentioned above 2 groups were collected, the TfR2 mRNA expression in BMMNC of patients with HA was detected by fluorescence-quantitative PCR, the correlation of HA with bone marrow erythroid hyperplasia, iron status of body and underlying diseases was analyzed. The results showed that the relative level of TfR2 mRNA expression in HA patients was significantly higher than that in control patients. The TfR2 mRNA expression level negatively correlated with Hb level in peripheral blood (r = -0.715), while it positively correlated with ratio of bone marrow erythroblasts (r = 0.533). It is concluded that TfR2 mRNA expression in HA patients increases and closely correlates with hyperplasia status of bone marrow and anemia level in peripheral blood.

Leukemic stromal hematopoietic microenvironment negatively regulates the normal hematopoiesis in mouse model of leukemia / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Leukemic microenvironment has a major role in the progression of leukemia. Leukemic cells can induce reversible changes in microenvironmental components, especially the stromal function which results in improved growth conditions for maintaining the malignant leukemic cells. This study aimed to investigate the survival advantage of leukemic cells over normal hematopoietic cells in stromal microenvironment in long term.</p><p><b>METHODS</b>The mice were injected intraperitoneally with N-N' ethylnitrosourea (ENU) to induce leukemia; the mice received injection of normal saline were used as control. At 180 days after ENU induction, the mice were killed and the bone marrows were cultured for 19 days. Colony-forming assays were used to analyze the formation of various cell colonies. The expression of Sca-1, CD146, VEGFR2, CD95, pStat3, pStat5, and Bcl-xL in marrow cells were detected by flow cytometry.</p><p><b>RESULTS</b>Long-term leukemic bone marrow culture showed abnormal elongated stromal fibroblasts with almost absence of normal hematopoietic cells. Adherent cell colonies were increased, but CFU-F and other hematopoietic cell colonies were significantly decreased in leukemia group (P<0.001). Primitive progenitor-specific Sca-1 receptor expression was decreased with subsequent increased expression of CD146 and VEGFR-2 in leukemic bone marrow cells. Decreased Fas antigen expression with increased intracellular pStat3, pStat5 and Bcl-xL proteins were observed in leukemic bone marrow cells.</p><p><b>CONCLUSIONS</b>Stromal microenvironment shows altered morphology and decreased maturation in leukemia. Effective progenitor cells are decreased in leukemia with increased leukemia-specific cell population. Leukemic microenvironment plays a role in promoting and maintaining the leukemic cell proliferation and survivability in long term.</p>

Doenças hematológicas associadas ao eritrovírus / Hematologic diseases associated with eritrovírus

O eritrovírus infecta células precursoras eritroides, determinando a interrupção temporária da eritropoese. Neste contexto, é importante o conhecimento das principais doenças hematológicas que podem estar associadas à presença do vírus, principalmente quando estão presentes em condições mórbidas, tais como nas anemias hemolíticas hereditárias. Este trabalho tem como objetivo relatar as principais doenças hematológicas que cursam com a infecção pelo eritrovírus B19.

Erythroviruses infect precursor erythroid cells, determining a temporary disruption of erythropoiesis. Thus, knowledge of the main hematological diseases that may be associated with the virus is important, especially when they are present in morbid conditions, such as in hereditary hemolytic anemia. This paper aims at reporting the main hematological diseases that are associated with erythrovirus infections.

Fuzzy mathematic quantitative law of composing principle in the study of traditional Chinese medicine / 药学学报

This study is to analyze microcosmic significance of Chinese medicine composing principle "principal, assistant, complement and mediating guide" and it's fuzzy mathematic quantitative law. According to molecular biology and maximal membership principle, fuzzy subset and membership functions were proposed. Using in vivo experiment on the effects of SiWu Decoction and its ingredients on mice with radiation-induced blood deficiency, it is concluded that DiHuang and DangGui belonged to the principal and assistant subset, BaiShao belonged to the contrary complement subset, ChuanXiong belonged to the mediating guide subset by maximal membership principle. It is discussed that traditional Chinese medicine will be consummate medical science when its theory can be described by mathematic language.