Neuroprotective effects of erythropoietin on rat retinas subjected to oligemia

OBJECTIVES: Erythropoietin may have neuroprotective potential after ischemia of the central nervous system. Here, we conducted a study to characterize the protective effects of erythropoietin on retinal ganglion cells and gliotic reactions in an experimentally induced oligemia model. METHODS: Rats were subjected to global oligemia by bilateral common carotid artery occlusion and then received either vehicle or erythropoietin via intravitreal injection after 48 h; they were euthanized one week after the injection. The densities of retinal ganglion cells and contents of glial fibrillary acidic protein (astrocytes/Müller cells) and cluster of differentiation 68 clone ED1 (microglia/macrophages), assessed by fluorescence intensity, were evaluated in frozen retinal sections by immunofluorescence and epifluorescence microscopy. RESULTS: Retinal ganglion cells were nearly undetectable one week after oligemia compared with the sham controls; however, these cells were partially preserved in erythropoietin-treated retinas. The contents of glial fibrillary acidic protein and cluster of differentiation 68 clone ED1, markers for reactive gliosis, were significantly higher in retinas after bilateral common carotid artery occlusion than those in both sham and erythropoietin-treated retinas. CONCLUSIONS: The number of partially preserved retinal ganglion cells in the erythropoietin-treated group suggests that erythropoietin exerts a neuroprotective effect on oligemic/ischemic retinas. This effect could be related to the down-modulation of glial reactivity, usually observed in hypoxic conditions, clinically observed during glaucoma or retinal artery occlusion conditions. Therefore, glial reactivity may enhance neurodegeneration in hypoxic conditions, like normal-tension glaucoma and retinal ischemia, and erythropoietin is thus a candidate to be clinically applied after the detection of decreased retinal blood flow.

Effect of emodin on rat bone marrow mesenchymal stem cell proliferation and mRNA expressions of hematopoietic growth factors / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of emodin on the proliferation, cell cycle distribution, apoptosis and expression of hematopoietic growth factors in bone marrow mesenchymal stem cells (BMSCs).</p><p><b>METHODS</b>The proliferation of rat BMSCs exposed to emodin was analyzed using MTT assay, and flow cytometry was used to detect the apoptosis and cell cycle changes of the exposed cells. Real-time quantitative PCR was used to determine the mRNA expression of the hematopoietic growth factors.</p><p><b>RESULTS</b>Exposure to 0.1 and 1 µg/ml emodin for 48 and 72 h significantly enhanced the proliferation of BMSCs (P<0.01). The cells exposed to 0.1 µg/ml emodin showed significantly increased percentage of cells in G2/M phase (P<0.05), and 1 µg/ml emodin exposure caused increased cells in S phase (P<0.01) and decreased cells in G1/G0 phase (P<0.05). Emodin exposure for 48 h resulted in significantly decreased cell apoptosis (P<0.05). BMSCs treated with 0.1 µg/ml emodin showed a significant increase in the expression of thrombopoietin mRNA (P<0.05).</p><p><b>CONCLUSION</b>Emodin can promote the proliferation of BMSCs in vitro possibly by regulating the cell cycle distribution, cell apoptosis and thrombopoietin expression.</p>

Actualización en trasplante de células progenitoras hematopoyéticas en pacientes pediátricos en los últimos 15 años / Updating in transplant of hematopoietic progenitor cells in pediatric patients during the past 15 years

En el pasado año 2010 se conmemoró el 25º aniversario de la introducción en Cuba del trasplante de médula ósea, y su desarrollo ha seguido la secuencia de la historia universal del trasplante hematopoyético. En este trabajo nos referimos a los logros más importantes que se han alcanzado en los últimos 15 años, como ha sido la introducción del trasplante con células movilizadas hacia la sangre periférica. Se exponen los resultados parciales de un estudio comparativo de 2 grupos de pacientes pediátricos, uno que recibió células progenitoras hematopoyéticas obtenidas de médula ósea y otro con células movilizadas hacia la sangre periférica mediante factores de crecimiento hematopoyéticos. Otros avances han sido: la introducción del trasplante no mieloablativo en el año 2002, la aplicación de factores recombinantes producidos en Cuba en el manejo de los pacientes trasplantados, y la introducción de técnicas de quimerismo. Se analizan diferentes aspectos relacionados con la histocompatibilidad y los requerimientos para mejorar los resultados del trasplante. Se señala la contribución que ha tenido la experiencia obtenida con este proceder, para el desarrollo de la medicina regenerativa

In the past year 2010, it was commemorate the 25 Anniversary of introduction in Cuba of the bone marrow transplantation and its development has followed the sequence of the universal history of the hematopoietic transplantation. In present paper authors made reference to more important achievements over the past 15 years including the introduction of the transplantation with mobilized cell to peripheral blood. Partial results of a comparative study of 2 groups of pediatric patients are showed; one received hematopoietic progenitor cells obtained from the bone marrow and other with cells mobilized to the peripheral blood by means of hematopoietic growth factors. Other advances include: the introduction of non-myeloablation transplantation in 2002, the application of recombinant factors produced in Cuba in the management of transplanted patients and the introduction of chimerism. Different features related to histocompatibility are analyzed as well as the requirements to improve the transplantation results. It is indicated the contribution of the experience obtained with this procedure for the development of the regenerative medicine

Comparative study of enhancing effect on mRNA expression of hematopoietic growth factors in rat bone marrow mesenchymal stem cells by ginseng polysaccharide and ginsenoside / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study mRNA expression levels of main hematopoietic growth factors in bone marrow mesenchymal stem cells (BM-MSC), and to compare effect on mRNA expression levels treated by ginseng polysaccharide and ginsenoside.</p><p><b>METHODS</b>Relative quantification real-time polymerase chain reaction (RT-PCR) was used to observe mRNA expression levels of IL4, Csf2, Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo, and IL3, etc. in rat BM-MSC treated with ginseng polysaccharide (20 microg/mL) or ginsenoside (20 microg/mL) at 12, 24, and 36 h.</p><p><b>RESULTS</b>IL4 and Csf2 mRNA expressions were not detected. Relative expression of Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo and IL3 mRNA ranked in an attenuating order when compared with Gapdh mRNA. mRNA expression of Epo and IL3 was not significantly changed at any time point by treatment of ginseng polysaccharide or ginsenoside in rat BM-MSC (P > 0.05). mRNA expression of Csf1, IL6, Lif, Csf3 and IL11 were significantly enhanced at 12 and 36 h by treatment of ginseng polysaccharide (P < 0.05) and that of Csf1, IL6, Lif, Csf3, and Kitlg were significantly enhanced at 24 h in rat BM-MSC (P < 0.05). The enhanced mRNA expression was Csf3 at 12 h, Csf3, IL6 and Lif at 24 h, and Csf3, IL6, Lif, IL11, and Kitlg, respectively at 36 h by treatment of ginsenoside in rat BM-MSC.</p><p><b>CONCLUSIONS</b>The enhancement of ginseng polysaccharide was stronger than that of ginsenoside on mRNA expression of hematopoietic growth factors in the initial stage. As time went by, the enhancement of ginsenoside gradually increased and exceeded that of ginseng polysaccharide.</p>

Auto-mobilized adult hematopoietic stem cells advance neovasculature in diabetic retinopathy of mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hematopoietic stem cells (HSCs) can be used to deliver functionally active angiostatic molecules to the retinal vasculature by targeting active astrocytes and may be useful in targeting pre-angiogenic retinal lesions. We sought to determine whether HSC mobilization can ameliorate early diabetic retinopathy in mice.</p><p><b>METHODS</b>Mice were devided into four groups: normal mice control group, normal mice HSC-mobilized group, diabetic mice control group and diabetic mice HSC mobilized group. Murine stem cell growth factor (murine SCF) and recombined human granulocyte colony stimulating factor (rhG-csf) were administered to the mice with diabetes and without diabetes for continuous 5 days to induce autologous HSCs mobilization, and subcutaneous injection of physiological saline was used as control. Immunohistochemical double staining was conducted with anti-mouse rat CD31 monoclonal antibody and anti-BrdU rat antibody.</p><p><b>RESULTS</b>Marked HSCs clearly increased after SCF plus G-csf-mobilization. Non-mobilized diabetic mice showed more HSCs than normal mice (P=0.032), and peripheral blood significantly increased in both diabetic and normal mice (P=0.000). Diabetic mice showed more CD31 positive capillary vessels (P=0.000) and accelerated endothelial cell regeneration. Only diabetic HSC-mobilized mice expressed both BrdU and CD31 antigens in the endothelial cells of new capillaries.</p><p><b>CONCLUSION</b>Auto-mobilized adult hematopoietic stem cells advance neovasculature in diabetic retinopathy of mice.</p>

Detecção da carga viral dos herpesvirus HHV-5 (citomegalovirus) e HHV-6 pela reação em cadeia da polimerase em tempo real e transcrição reversa acoplada a nested-PCR em pacientes receptores de transplante de celulas tronco hematopoieticas / Detection of herpesvirus HHV-5 (cytomegalovirus) and HHV-6 viral load by real time polymerase chain reaction and reverse transcription nested polymerase chain reaction in hematopoietic stem cell transplantation recipients

O cytomegalovirus humano (HCMV) e o herpesvirus humano 6 (HHV-6) são β-herpesvirus com homologia superior a 67 por cento e alta soroprevalência na população adulta. A infecção primaria por estes herpesvirus ocorre comumente na infância e é normalmente subclinica, ou pode causar mononucleose (HCMV) ou exantema súbito (HHV-6) sendo resolvidos na maioria dos casos sem complicações. Após a infecção primária os vírus permanecem no hospedeiro por toda vida podendo ser reativado de seu estado de latência em indivíduos adultos imunocomprometidos como os receptores de células tronco hematopoiéticas (TCTH). A reativação ou reinfecção por estes vírus causam serias complicações em pacientes submetidos ao transplante de células tronco hematopoiéticas como pneumonia intersticial, febre, gastroenterite, mielossupressão, encefalite e doença do enxerto contra o hospedeiro (GVHD). A reativação do HHV-6 após o transplante é associada com o desenvolvimento de infecções oportunistas, doença causada pelo citomegalovírus humano e possíveis episódios de rejeição aguda. Com efetivos tratamentos antivirais disponíveis, um monitoramento adequado destes vírus distinguindo entre latência e reativação é critico para estes pacientes. Monitoramos 30 pacientes submetidos à TCTH quanto a infecção ativa por HCMV e HHV-6 pelas técnicas de nested-PCR em soro e células, PCR- em tempo real em soro e células e transcrição reversa acoplada a nestedPCR (RT-nPCR). 29 pacientes (96,66 por cento) apresentaram infecção ativa por HCMV sendo 21 pacientes (70 por cento) pela nested-PCR em células, 17 pacientes(56,66 por cento) pela neste-PCR em soro, 23 pacientes(76,67 por cento) pela PCR em tempo real em células,19 pacientes (63,33 por cento) pela...

Human cytomegalovirus (HCMV) and human herpesvirus type 6 (HHV-6) are β-herpesvirinae extremely closely related with a homology > 67 per cent with a high seroprevalence in the adult population. Primary infection commonly appears in early childhood and is usually subclinical, or may cause mononucleosis (HCMV) or febrile illness, including exanthema subitum (HHV-6), solving, in the majority of cases, without complications. After primary infection, the viruses persist in the infected individual through life and can be reactivated from their state of latency in immunocompromised hosts. Reactivation or reinfection causes severe clinical diseases in patients who underwent hematopoietic stem cell transplantation, like interstitial pneumonia, fever, gastroenteritis, myelossupression, encephalitis and graft-versus-host-disease (GVHD). A potential increase in virulence of HHV-6 in the course of a simultaneous CMV reactivation, leading to a great risk of CMV-associated disease. In this present study, 30 patients who received HSCT were monitoring for active HCMV and HHV-6 infection by Nested PCR in serum and peripheral blood leukocytes (PBL) samples, real time PCR in serum and PBL and RT-nPCR. In 29 patients (96,66 per cent) active HCMV infection was detected: 21 patients (70 per cent) by PBL nested-PCR, 17 patients (56,66 per cent) by serum neste-PCR, 23 patients (76,67 per cent) by PBL nested real-time-PCR, 19 patients (63,33 per cent) by...

Assessment the effectiveness of hematopoietic growth factors on hepatic regeneration after partial hepatectomy in albino rats with induced bone marrow suppression

This study aimed to evaluate the effect of hematopoietic growth factors [erythropoietin and granulocyte colony stimulating factor] on liver regeneration after partial hepatectomy and bone marrow suppression of rats and to investigate the role of hematopoietic stem cells in liver regeneration by assessment of CD 34+ hematopoietic stem cells marker in hepatic tissues. 36 adult male albino rats were used in this study and were divided into six groups each of 6 rats: the 1st is the control group, the 2nd is bone marrow suppression group, 3rd is subjected to partial hepatectomy, 4th is subjected to bone marrow suppression by benzene, then 70% partial hepatectomy, 5th is subjected to 70% partial hepatectomy with injection of Eprex and Neupogen at time of partial hepatectomy, then daily for 5 days post hepatectomy and the 6th group is subjected to bone marrow suppression by benzene first then 70% partial hepatectomy with injection of Eprex and Neupogen daily for 5 days post hepatectomy. Histological evaluation and immunohistochemical study for CD34+ cells in the hepatic tissues were assessed. There were no regenerative changes in both control and benzene treated groups. There were little regenerative changes in the group of partial hepatectomy after benzene treatment but in the groups that were treated with the hematopoietic growth factors. These regenerative changes were increased especially in the treated group after partial hepatectomy than treated group after partial hepatectomy and bone marrow suppression. Immunostaining of CD34 expression as marker of HSCs in liver sections showed the following; the normal group, benzene treated group and partially hepatectomized group after benzene treatment showed no expression, but partially hepatectomized group show focally positive expression. Both groups treated with hematopoietic growth factors either after partial hepatectomy or after partial hepatectomy and bone marrow suppression by benzene show diffusely positive expression. The present study had shown that Erythropoietin and Granulocyte colony stimulating factors stimulate regenerative process occurred in the liver after partial hepatectomy only and after partial hepatectomy with bone marrow suppression either by endogenous mechanisms or by mobilization of hematopoietic stem cells

Role of granulocyte colony-stimulating factor [G-CSF]/Filgrastim as an adjunct in chronic hepatitis C management

Drug-induced hematotoxicity is the commonest reason for reducing the dose or withdrawing interferon [IFN] therapy in a case of chronic hepatitis C thus depriving the patient of a possible cure. Traditionally, severe neutropenia has been considered an absolute contraindication to start antiviral therapy. Since the advent of adjunct therapy with Granulocyte-colony stimulating factor, the same is not true any more. Some recent landmark studies have used this adjunct therapy to help avoid antiviral dose reductions. although, addition of this adjunct therapy has been shown to significantly increase the overall cost of the treatment, if the infection is cured at the end of the day, this extra cost is worth bearing. Although, more studies are needed to refine the true indications of this adjunct therapy, determine the best dose regimen, quantify the average extra cost and validate that whether or not the addition of this therapy increases the sustained virologic response rates achieved, the initial reports are encouraging. Therefore, although not recommended on routine basis, some selected patients may be given the benefits of these factors. In this article, a review of the current literature on this subject is given followed by few suggested recommendations at the end to help develop local guidelines

Have hematopoietic growth factors made an impact on the management of liver disease?

It is clear that the major indication for the use of hematopoietic growth factors in hepatology is to counteract the adverse effects of interferons (neutropenia and thrombocytopenia) and ribavirin (hemolytic anaemia) during the treatment of hepatitis C infection. This is important because the probability of SVR depends on proper adherence to therapy (at least 80% of the requisite dose maintained for at least 80% of the requisite duration) and proper adherence can only be achieved if the side effects are reduced to a minimum. Even though the studies have demonstrated beyond doubt that the use of hematopoietic growth factors does indeed reduce the incidence and severity of these adverse effects and helps the patients to complete the course of therapy, the data on improvement of SVR is still limited. There is only one study of darbepoetin and filgrastim showing the beneficial effect on SVR. Even among the hematological side effects, possibly the only significant effect which limits the use of optimal HCV therapy is the hemolytic anaemia induced by ribavirin. The other two main side effects, i.e. neutropenia and thrombocytopenia are not clinically problematic. The use of such growth factors would be particularly effective if patients who have advanced liver disease or cirrhosis are able to receive adequate anti-viral therapy as has been demonstrated in the study of eltrombopag among HCV cirrhotics. Apart from this, other indications of G-CSF or GM-CSF use are still in the experimental stage. So, as of now, apart from erythropoietic factors, the role played by other hematopoietic growth factors in hepatology is limited. But future research, especially in the areas of immunotherapy of liver cancers and stem cell therapy for endstage liver disease, is surely going to give these factors their due place in hepatology.

O uso profilático de fatores de crescimento, estimuladores de colônias hematopoéticas, em pacientes oncológicos submetidos à quimioterapia / The prophylactic use of growth factors, stimulating of Hematopoietic colonies, in oncological patients submitted to the chemotherapy

O objetivo dos autores é fazer uma revisão sobre as indicações atuais do uso de fatores de crescimento, estimuladores de colônias hematopoéticas, na profilaxia da neutropenia febril, em pacientes submetidos à quimioterapia, avaliando seu custo-benefício.

Effect of hematopoietic stimulating factors on the expansion of megakaryocyte / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of hematopoietic stimulating factors on the expansion of mature megakaryocytes.@*METHODS@#(2, 4, 6, 8, 10) x 10(5)/mL bone marrow single nucleus cells (BMNC) were added in the culture system of colony forming unit-megkaryocyte (CFU-Meg) to find out the relationship of the cultured BMNC with the output of CFU-Meg. rmSCF + rmTPO + rmIL-3 (3HSFs) and rmSCF + rmTPO + rmIL-3 + rmIL-6 (4HSFs) or F-CM were added in the liquid culture system of megkaryocytes respectively. The number of mature megakaryocytes were counted every other day.@*RESULTS@#The number of CFU-Meg increased with the increase of the cultured BMNC. The CFU-Meg productivity of 1 x 10(6) BMNC/mL culture system was more than that of 2 x 10(5) BMNC/mL culture system. 3HSFs and 4HSFs or F-CM significantly promoted the expansion of mature megakaryocytes in the liquid culture system, but the effect was different. The peak time of the number of mature megakaryocytes in 3HSFs and 4HSFs or F-CM were 7 d, 7 d and 5 d respectively.@*CONCLUSION@#3HSFs and 4 HSFs or F-CM had positive effect on the expansion of mature megakaryocytes. 4HSFs was better than 3HSFs and F-CM. 3HSFs was better than F-CM. The peak time of the number of mature megakaryocytes in different culture systems was different.

The response of bone marrow hematopoietic cells to G-CSF in paroxysmal nocturnal hemoglobinuria patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the response of hematopoietic cells (HSC) to granulocyte colony stimulating factor (G-CSF) in paroxysmal nocturnal hemoglobinuria (PNH) patients.</p><p><b>METHODS</b>(1) Bone marrow mononuclear cells (BMMNC) from 17 PNH patients and 12 normal subjects were inoculated into semisolid culture media containing or not G-CSF (50 ng/ml). The cluster/colony forming unit-granulocyte/monocyte (CFU/cFU-GM) were counted and compared. (2) BMMNC of 20 PNH patients and 12 normal controls were triply stained for CD34, CD59 and G-CSF receptor CD114/stem cell factor receptor (C-KIT) CD117 and assessed by FCM. The CD34(+) cells were identified as CD34(+)/CD59(+) and CD34(+)/CD59(-). Percentage of CD114 and CD117 expression in each cell population was calculated.</p><p><b>RESULTS</b>(1) PNH cFU-GM without G-CSF were (112.41 +/- 22.74)/10(5) BMMNC, while with G-CSF: (133.82 +/- 25.85)/10(5) BMMNC and normal cFU-GM were (190.33 +/- 36.05)/10(5) BMMNC, (309.42 +/- 92.94)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC formed less cFU-GM than control did, both of the two kinds of BMMNC responded to G-CSF well (P < 0.05), but the increment of PNH cFU-GM yields was less than that of the normal control (P < 0.05). CFU-GM yields of PNH BMMNC without G-CSF were (24.29 +/- 9.05)/10(5) BMMNC, with G-CSF were (27.53 +/- 10.65)/10(5) BMMNC, while normal control were (77.42 +/- 36.01)/10(5) BMMNC and (98.00 +/- 43.14)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC showed less CFU-GM yields than that of control (P < 0.05). (2) The percentage of CD114 positive cells in PNH CD34(+)CD59(+) BMMNC was (73.34 +/- 29.40)% and that in PNH CD34(+)CD59(-) BMMNC and in control CD34(+)CD59(+) BMMNC were (32.70 +/- 6.89)% and (58.52 +/- 29.99)%, respectively. The percentage of CD114 expression in PNH CD34(+) CD59(-) BMMNC was less than that in the other two groups (P < 0.05). The percentages of CD117 positivities on the PNH CD34(+)CD59(+) BMMNC were (76.90 +/- 22.08)%, PNH CD34(+) CD59(-) (36.03 +/- 7.69)% and control CD34(+) CD59(+) (80.28 +/- 13.36)%, respectively (P < 0.01).</p><p><b>CONCLUSION</b>In vitro, BMMNC of normal control grow better, and respond better to G-CSF than PNH BMMNC do. PNH CD34(+)CD59(-) BMMNC express less G-CSF receptor and C-KIT than PNH CD34(+)CD59(+) and normal CD34(+)CD59(+) BMMNC do, which may be the reason that abnormal PNH clone grow worse than the normal clones do.</p>

Effects of human umbilical cord blood mesenchymal stem cells on the expansion of CD34+ cells from umbilical cord blood / 中华儿科杂志

<p><b>OBJECTIVE</b>The previous studies indicated that mesenchymal stem cells (MSCs) either from umbilical cord blood (UCB) or from bone marrow (BM) had the same biological characteristics and the function of secreting hematopoietic growth factors (HGFs). The present study aimed to understand the effects of human UCB MSCs on the expansion of CD(34)(+) cells from UCB.</p><p><b>METHODS</b>1. Human UCB CD(34)(+) cells were incubated in the system containing UCB MSCs, HGFs and serum free medium. 2. The surface markers (CD(34)(+), CD(34)(+)CD(38)(-), CD(34)(+)CD(3)(+), CD(34)(+)CD(19)(+), CD(34)(+)CD(33)(+), CD(34)(+)CD(41a)(+)) on expanded UCB cells were examined by flow cytometry on the 6th and 12th days. 3. The expanded and unexpanded cells were cultured in semi-solid culturing system and checked for colony forming units of granulocyte and macrophage (CFU-GM), erythroid burst-forming unit (BFU-E), colony forming units of granulocyte- erythrocyte-megakaryocyte-macrophage (CFU-Mix) and colony forming units of high-proliferative potential (CFU-HPP).</p><p><b>RESULTS</b>1. The expansion folds of CD(34)(+)CD(38)(-) cells from UCB MSCs + HGFs groups on the 6th and 12th days were 159.43 and 436.68, respectively. Interestingly, the percentage of CD(34)(+)CD(38)(-) cells declined in HGFs group after expanding for 12 days, but it rose to 9.98% in the UCB MSCs + HGFs group. 2. Colony forming capacity of expanded UCB cells showed that the folds of CFU-Mix and CFU-HPP of UCB MSCs + HGFs group increased from day 6 to day 12, but the folds decreased in the HGFs group. 3. From day 0 to day 12, CD(34)(+)CD(33)(+) cells and CD(34)(+)CD(41a)(+) cells were amplified gradually, but CD(34)(+)CD(19)(+) and CD(34)(+)CD(3)(+) cells decreased gradually, and in UCB MSCs + HGFs group this phenomenon was more significant than that in HGFs group.</p><p><b>CONCLUSION</b>1. UCB MSCs containing system not only has the ability to expand the primitive HSCs but also has the ability to sustain the proliferation of HSCs. 2. UCB MSCs containing system amplified mainly myeloid and megakaryocytoid progenitor subsets. These may have clinical significance in reducing infection and hemorrhage.</p>

Study on the expression of hematopoietic growth factor gene in human umbilical vein endothelial cells using gene chip / 中国实验血液学杂志

In order to detect the hematopoietic growth factor gene expressed in human umbilical vein endothelial cells using gene chip, human umbilical vein endothelial cells (ECV304) were cultured in vitro and divided into VEGF group and control group in same medium. 50 ng/ml hVEGF165 was added in the VEGF group. After culture for 24 hours all cells were collected for total RNA extraction. Then, cDNAs were marked with Cy3 and Cy5 for control group and VEGF group, respectively, and hybridized with gene chip. Hybridization signals were collected and analyzed following scanning by laser co-focal microscopy. The results showed that a large number of hematopoietic growth factor and receptor genes (Epo/R, GM-CSF/R, G-CSF/R, LIF, IL-3, TPO, Flt-3, SCF) were expressed in both groups, while many other growth factors (VEGF, IGF2, PDGFA, PDGFB, TGFbeta1) and receptors (neuropilin-1, neuropilin-2, TGFbeta-R1)were expressed. The differentially expressed genes amounted to 24. It is concluded that many hematopoietic growth factors and receptors expressed by hUVECs could be analyzed in a short period by using gene chip, which provides a powerful method for further studies on characteristics of vascular endothelial cells.

Anemia aplástica: experiencia con 7 casos / Aplastic anemia: a report of 7 cases

La anemia aplástica (AA) es una falla medular caracterizada por pancitopenia en sangre periférica como resultado de una disminución de la producción de células sanguíneas en médula ósea. Tiene diversas etiologías y una incidencia de 2 a 4 casos por 1 000 000 niños menores de 15 años. El tratamiento de elección es el transplante de médula ósea alogénico o en su defecto la inmunosupresión con lingoglobulina o timoglobulina además de ciclosporina, metilprednisolona y factores de crecimiento hematopoyético. Se presenta la experiencia con 7 pacientes del Hospital Roberto del Río diagnosticados entre los años 1995 y 2000, edad 2 meses a 13 años, con biopsia de médula ósea compatible. Dos pacientes presentaban etiología congénita, 3 con antecedentes de hepatitis y 2 fueron considerados idiopáticos. Un paciente fue transplantado de un hermano compatible luego de recibir inmunosupresión, 3 recibieron inmunosupresión con linfo/timoglobulina además de ciclosporina y factor estimulante de colonias de granulocitos y 3 niños sólo recibieron tratamiento de sostén con metilprednisolona o factores de crecimiento. Dos pacientes fallecieron precozmente por cuadro infeccioso. Cinco pacientes están vivos con una mediana de seguimiento de 43 meses, los 4 que recibieron inmunosupresión incluido el paciente transplantado, y la paciente con anemia de Fanconi

Células mononucleares de sangue de cordão umbilical e de sangue periférico estimulado com fator de crescimento granulocítico (G-CSF): análise da proliferação e de apoptose in vitro / Mononuclear cells of umbilical cord blood and of granulocyte colony-stimulating factor: analyses of proliferation and apoptosis in vitro

Células mononucleares de sangue de cordão umbilical (SCU) e sangue periférico mobilizado (SPM) com G-CSF, foram cultivadas in vitro com citocinas, na presença ou não de estroma de medula óssea. Os objetivos foram avaliar a capacidade proliferativa de células progenitoras, a ocorrência de apoptose e expressão de integrina. Nas culturas sem estroma, a celularidade aumentou 5 vezes (SCU) e não se alterou nas de SPM. O total de células CD34+ caiu em ambas culturas. Com estroma, o total de células nucleadas aumentou 7 vezes (SCU) e 2,3 vezes (SPM). O total de células CD34+ permaneceu o mesmo. A apoptose foi menor nas culturas de SCU. A expressão de integrina caiu, na população de células CD34+ e de CD45+ / Mononuclear cells from umbilical cord blood (UCB) and G-CSF mobilized peripheral blood (MPB), were cultured in vitro, in the presence of cytokines, with or without bone marrow stroma. The aims were to evaluate the proliferative response of progenitor cells, occurrence of apoptosis and expression of adhesion molecule. In cultures without stroma, cellularity increased 5-fold for UCB, but has not changed for MPB. The number of CD34+ cells has dropped in both culture. With stroma, total nucleated cells had a 7-fold increse (UCB) and a 2,3-fold (MBP), however, CD34+ cells number has not changed. Apoptosis was lower in UCB culture. The expression of integrin decreased, in the...

Avaliação da presença e atividade biológica de citocinas hematopoiéticas no colostro humano / Evaluation of the presence and biological activity of hematopoietics cytokines in colostrum human

Técnicas foram aplicadas para a detecção de citocinas: por imunocitoquímica para as citocinas; por marcação intracelular em citometria de fluxo para as citocinas IL-4, IL-5,IL-6,INF-y e GM-CSF. As células do colostro são constituídas principalmente por macrófagos e lifócitos. A representação destas populações foi confirmada pela distribuição dos marcadores de superfície CD11b e CD3. A marcação intracelular com CD14 forneceu resultados comparáveis aos da marcação de superfície com CD11b. Em contraste, a freqüência de células com positividade para CD14 na superfície foi muito baixa, o que foi interpretado, como evidência de rápida conversão do CD14 de membrana em CD14 solúvel. As células T do colostro incluem subpopulações com marcadores fenotípicos de ativação CD69,e subpopulações com marcadores fenotípicos de células de memória CD45RO. Foram encontradas células com marcação para o CD158, indicando a presença de células NK. Evidenciou presença das citocinas imuno-regulatórias IL-4, IL-5 em microscopia ótica e IL-4, IL-5 e INF-y, por detecção intracitoplasmática em citometria de fluxo. Estes resultados permitiram caracterizar a presença de células Th1 e Th2 no colostro. Foram igualmente detectadas IL-10, TNF-a e GM-CSF, e associadas a macrófagos. Evidenciou a presença de fatores hematopoiéticos biologicamente ativos utilizando como bio-ensaio a indução de proliferação e a manutenção da viabilidade celular em culturas líquidas de células hematopoiéticas do sangue de cordão umbilical. Colostro teve atividade superior à da associação GM-CSF/IL-3 num período de 7-14 dias de cultura. A atividade biológica foi perdida progressivamente com períodos crescentes. A transferência de um número expressivo de células da mãe para o recém-nascido com capacidade de secretar citocinas biologicamente ativas, a presença de linfócitos T de memória, e o estado ativado em que muitas células se encontram, além das atividades biológicas demonstradas ex vivo, são compatíveis com a hipótese de que o colostro exerce um importante papel na imunomodulação da criança. Os resultados apontam, em estudos futuros: os alvos da ação das citocinas detectadas, no próprio colostro e/ou no revestimento do trato digestivo; possíveis mecanismos pelos quais a transferência adotiva de imunidade inata e adquirida via colostro possa exercer efeitos sistêmicos no lactente.

Human bone marrow mesenchymal stem cells express multiple hematopoietic growth factors / 中国实验血液学杂志

To study the biological role of human cultured bone marrow mesenchymal stem cell (BM-MSC) in hematopoiesis by investigation of its expression of multiple hematopoietic growth factors, RT-PCR was used to analyze the expression of SCF, Flt3-ligand, TPO, LIF, G-CSF, GM-CSF, IL-3, IL-6 and IL-11 at mRNA level for human BM-MSC from healthy donors and patients with leukemia and lymphoma. BM-MSC were incubated with or without hydrocortison (HC). The results clearly showed that the cultured BM-MSC expressed mRNA of SCF, Flt3-ligand, TPO, LIF, IL-6 and IL-11 at passages 3 up to 15, but did not express G-CSF, GM-CSF and IL-3. The same expression pattern of above cytokines was seen also for the patient's BM-MSC. HC was able to induce BM-MSC to express G-CSF but not to express GM-CSF. BM-MSC seemed not to change morphologically after incubation with HC for up to 21 days. In conclusion, both normal and patient BM-MSC should be potential to promote hematopoiesis according to their expression of multiple hematopoietic cytokines, and HC is able to induce hematopoietic growth factor expression.

Expression of survivin in cord blood CD34(+) stem/progenitor cells and its significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression and significance of survivin protein and mRNA in cord blood (CB) CD(34)(+) stem/progenitor cells.</p><p><b>METHODS</b>Survivin level in CB CD(34)(+) cells was assessed by immuno-histochemistry and reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Survivin expressed in fresh CB CD(34)(+) cells, its protein level was 9.4 +/- 1.2, faint signal of the mRNA transcript band was detected. In the in vitro culture of the cells, hematopoietic growth factors could upregulate survivin expression, especially the combination of SCF, FL and Tpo. After 3 days culture, its protein level was 25.2 +/- 5.3, and mRNA transcript increased significantly. Survivin expression dropped down 24 hours after withdraw of these cytokines.</p><p><b>CONCLUSION</b>Survivin is not a specific anti-apoptotic protein of cancer, and also expressed in normal hematopoietic stem/progenitor cells. It plays important regulating roles in the process of hematopoiesis.</p>