Protective effects of deferasirox and N-acetyl-L-cysteine on iron overload-injured bone marrow

Using an iron overload mouse model, we explored the protective effect of deferasirox (DFX) and N-acetyl-L-cysteine (NAC) on injured bone marrow hematopoietic stem/progenitor cells (HSPC) induced by iron overload. Mice were intraperitoneally injected with 25 mg iron dextran every 3 days for 4 weeks to establish an iron overload (Fe) model. DFX or NAC were co-administered with iron dextran in two groups of mice (Fe+DFX and Fe+NAC), and the function of HSPCs was then examined. Iron overload markedly decreased the number of murine HSPCs in bone marrow. Subsequent colony-forming cell assays showed that iron overload also decreased the colony forming capacity of HSPCs, the effect of which could be reversed by DFX and NAC. The bone marrow hematopoiesis damage caused by iron overload could be alleviated by DFX and NAC.

Counteracting the enzymatic activity of dipeptidylpeptidase 4 for potential therapeutic advantage, with an emphasis on cord blood transplantation

Dipeptidylpeptidase (DPP) 4, also known as CD26, is an enzyme present on the surface of a number of different cell types. It is also found within cells and as a soluble protein in body fluids. It can specifically truncate proteins at the penultimate N-terminus residue for some amino acids, such as alanine, proline, serine, and perhaps others. DPP4 has been implicated in regulating the in vitro and in vivo functional activities of a number of hematopoietically active molecules, and this information, along with that on inhibition of DPP4, has been studied in efforts to enhance hematopoietic cell transplantation (HCT), hematopoiesis after stress in mouse models, and in the clinical setting of single-unit cord blood (CB) HCT. This article reviews the current status of this compound's effects on regulatory proteins, the field of CB HCT, a potential role for modulating DPP4 activity in enhancing single-unit CB HCT in adults, and future aspects in context of other cellular therapies and the area of regenerative medicine.

Toxicity of trigonella foenum graecum [fenugreek] in bone marrow cell proliferation in rat

Fenugreek has a wide range of medical applications and its medicinal use has been clear in several studies, however, few studies are available on effects on haematopoietic stem cell of bone marrow. The goal of the present study was to investigate the effect of Fenugreek on fetal macroscopic diameters and microscopic bone marrow cell histological changes in its teratogenic dosages. Fenugreek decoction was dissolved in 1.5 milliliter distilled water and injected intraperitoneumly in three dosages of 0.8 g/kg, 1.6 g/kg, and 3.2 g/kg for three groups of Wistar female rats mated by Wistar male. For another group [as control group] only 1.5 milliliter distilled water was injected. Bone marrow tissue was prepared from rat fetus and was cut using a microtome and stained with hematoxylin and eosin. Sections were evaluated for changes using light microscope. LD50 for the measurement of teratogenic dosage of fenugreek was 4.1 and 3.5 g/kg in female and male rat, respectively. There was a positive relation between the injected drug dosage and fetal mortality rate. Among all fetal diameters, ear to ear diameter was decreased in groups received Fenugreek decoction. The severity of stem cell histological changes caused by 3.2 g/kg drug injection was lower than distilled water injection and in evaluation of other cells, differences in the severity of histological changes across three groups with different drug dosages and control group was detected. Fenugreek in teratogenic dosages can decrease the severity of bone marrow cell proliferation and increase fetal mortality rate

Niveles de citoquinas megacariocitopoyéticas en pacientes con trombocitemia esencial y su relación con características clínicas y bioquímicas / Megakaryopoietic cytokine levels in patients with essential thrombocythemia and their relationship with clinical and biochemical features

La megacariocitopoyesis y la producción de plaquetas están regidas por factores de transcripción y citoquinas presentes en el microambiente medular. La trombocitemia esencial (TE) es una enfermedad mieloproliferativa crónica caracterizada por aumento del recuento de plaquetas e hiperplasia megacariocítica. En el presente trabajo se evaluaron los niveles de las citoquinas que participan en el desarrollo megacariocítico en plasma de pacientes con TE que se encontraban sin tratamiento y los de trombopoyetina (TPO) antes y durante el tratamiento con anagrelide. Las determinaciones se realizaron por técnica de ELISA. Dentro de las citoquinas involucradas en la etapa de proliferación, los niveles de interleuquina 3 (IL-3) se encontraron aumentados en los pacientes (p=0.0383) respecto al grupo control. Los niveles de factor estimulante de colonias granulocito-macrofágico y stem cell factor fueron normales. Dentro de las citoquinas con acción sobre la maduración megacariocítica, tanto la interleuquina 6 como la interleuquina 11 y la eritropoyetina estuvieron normales. Los niveles de TPO antes del tratamiento no difirieron del grupo control y durante el tratamiento aumentaron de manera no significativa. Los pacientes que presentaron agregación espontánea tuvieron niveles más altos de TPO que los que no lo hicieron (p=0.049). Los niveles de las citoquinas no tuvieron relación con ninguno de los parámetros clínicos ni de laboratorio evaluados. El aumento de los niveles de IL-3 podría contribuir al incremento en la proliferación megacariocítica en este grupo. La presencia simultánea de niveles más altos de TPO y trombocitosis sería un factor predisponente para la ocurrencia de agregación espontánea en los pacientes con TE

Megakaryopoiesis and platelet production are driven by transcription factors and cytokines present in bone marrow environment. Essential thrombocythemia (ET) is a chronic myeloproliferative disorder characterized by high platelet count and megakaryocytic hyperplasia. In the present work we evaluated plasmatic levels of cytokines involved in megakaryocytic development in a group of patients with ET that were not on treatment, as well as thrombopoietin (TPO) levels before and during anagrelide treatment. The assays were carried out using ELISA techniques. Among the cytokines mainly involved in proliferation of megakaryocytic progenitors, interleukin 3 (IL-3) levels were found increased in patients compared to normal controls (p=0.0383). Granulocyte-macrophage colony stimulating factor and stem cell factor levels were normal. Interleukin 6, as well as interleukin 11 and erythropoietin (EPO), cytokines mainly related to megakaryocytic maturation, were normal. Plasma TPO levels before treatment were within the normal range and increased during treatment but the difference was not statistically significant. Patients who displayed spontaneous platelet aggregation had higher plasma TPO levels compared to those who did not (p=0.049). We did not find any relationship between cytokine levels and clinical or laboratory parameters. The high IL-3 levels seen in some patients with ET could contribute to megakaryocytic proliferation. The simultaneous occurrence of higher TPO levels and elevated platelet count could be a predisposing factor for the development of spontaneous platelet aggregation in ET patients

Comparative haematological and hepatorenal toxicity of IGR, lufenuron and profenofos insecticide on albino rats

The present investigation was conducted to compare the toxicity of the IGR, lufenuron and the organophosphorus insecticide, profenofos on blood content, liver and kidney functions of male albino rats. The tested compounds were orally administered to rats at 1/20 and 1/10 of their median lethal doses [LD50s] for two months [day after another], then toxicants were withdrawn for 30 days to allow recovery of toxic effects. Data indicated that 1/10 LD50 of both compounds caused significant changes on blood contents and biochemical parameters of treated rats without return to normal levels at the end of recovery period, while, the smallest dose revealed negligible changes on some tested parameters with resumed normal values. The adverse effects reached its peak at 45 and 60 days of treatment [high dose treated rats] followed by decrease during recovery intervals without returned to normal, but at 1/10 LD50, lufenuron caused sever damage on kidney; urea and creatinine showed high levels at the end of recovery periods [92.0 and 220.0% above normal level, respectively]. Data indicated that, 1/10 LD50 of lufenuron treated rats exhibited changes in leucocytes, platelets counts, transaminases activities, creatinine and urea concentrations more than the organophosphorus insecticide. On the contrary, the same dose of profenofos mostly affected on erythrocytes counts, haemoglobin levels and alkaline phosphatase [ALP] activity. The obtained data would suggest that the two tested compounds at high dose have an inhibitory action on haemopiesis. In addition, both compounds proved to have comparable toxicity towards animals

The effects of Ligustrazine on the expression of bFGF and bFGFR in bone marrow in radiation injured mice / 华中科技大学学报（医学）（英德文版）

To study the expression of the bFGF and its receptor in the mouse bone marrow by treatment with acute radioactive injury and Ligustrazine, 56 mice were divided into 3 groups: normal group, radiation-injured group and Ligustrazine group. After irradiation by 6.0 Gy 60Co gamma-ray, each mouse was orally given 0.1 ml Ligustrazine twice a day for 13 days in Ligustrazine group, and each mouse in radiation injured group was orally given equal amount of saline. On the 3rd, 7th, 14th day after irradiation, bone marrow mono-nuclear cells (BMMNC) were counted, and the expression levels of bPGF and bFGFR in bone marrow were evaluated by immunohistochemistry and flow cytometry analysis respectively. On the 3rd, 7th, 14th day after irradiation, expression of bFGF in bone marrow were significantly lower than in normal group (P<0.05 or P<0.01). Expressions of bFGF and bFGFR were much higher in Ligustrazine treated group than that in the control group (P<0.05 or P<0.01). Ligustrazine potentiate the expression of bFGF and bFGFR in bone marrow MNC to recover the bone marrow hematopoiesis inductive microenvironment, which is one of the mechanisms by which Ligustrazine rebuild the bone marrow hematopoiesis after acute radioactive injury.

Effects of ligustrazine on hematopoiesis in the early phase of bone marrow transplantation mice / 华中科技大学学报（医学）（英德文版）

To investigate the effects of Ligustrazine on histogenesis of bone marrow in the early phase of hematopoietic reconstruction in bone marrow transplantation (BMT) mice. The syngeneic BMT mice model was established. The syngeneic BMT mice were orally given 2 mg Ligustrazine twice a day. 1, 3, 5, 7, 10, 15 and 21 day(s) after BMT, peripheral blood granulocytes and bone marrow nucleated cells (BMNC) were counted and the diameter of central vein and the area of micro-vessel in femur were measured. The effect of Ligustrazine on hematopoietic stem cells was observed by colony forming unit of spleen (CFU-S). The effect of Ligustrazine on hemopoietic progenitors was studied by observing the number of progenitors of Granulocytes/Macrophage on day 10 and day 20 after BMT. In Ligustrazine-treated group, the diameter of center veins and the area of micro-vessel of femur were all significantly less than the control group 7, 10, 15, 21 days after BMT (P < 0.01). In addition, Ligustrazine significantly increased the number of CFU-S on day 10 and the number of CFU-GM on day 10, 20 after BMT. These results indicate that Ligustrazine can accelerate the histogenesis of hemopoietic bone marrow, which may be one mechanism by which Ligustrazine promotes hematopoietic reconstitution after BMT.

El caseinato de sodio induce la diferenciación de las células hematopoyéticas multipotenciales 32D / Sodium caseinate induces differentiation of the multipotential hemopoietic cell line 32D

Objetivo. Evaluar la participación del caseinato de sodio (CasNa) en la modulación de la hematopoyesis. Material y métodos. Se emplearon células 32D, una línea celular hematopoyética multipotencial, de origen murino y dependiente de interleucina-3. Estas células se cultivaron con 0.5 ng/mL de interleucina-3 y con concentraciones variables de CasNa. En los cultivos se realizaron estudios de proliferación celular (conteo directo e incorporación de timidina 3H) y de diferenciación morfológica (tinción con Giemsa), citoquímica (tinciones específicas para monocito-macrófagos y para granulocitos) y funcional (presencia de receptores Fc y reducción de nitro azul de tetrazolio), además se determinó la viabilidad con azul de tripano y la apoptosis por la reacción Tunel in situ. Resultados. Se demostró que el CasNa produce una reducción en la proliferación, dependiente de la dosis, que ésta no es provocada por una disminución de la viabilidad de las células 32D así como tampoco por un aumento de la muerte celular por apoptosis. Además el CasNa indujo la diferenciación de las células 32D hacia monocito-macrófagos en cultivos de 4 días. Conclusiones. Aparentemente el CasNa ejerce una actividad tipo factor estimulador de colonias de macrófagos. Además, parece ser un potente factor de diferenciación de las células 32D, ya que en sólo 4 días de estímulo genera células de tipo monocito-macrófago, a diferencia de los 7 días requeridos por la combinación de G-CSF y GM-CSF.

Evaluation of amlodipine in mild to moderate hypertension--a clinical report.

Twenty two patients having mild to moderate hypertension were treated with a single daily dose of amlodipine for 4 weeks. Satisfactory response defined as final diastolic blood pressure < 90 mm of Hg and a reduction from baseline values > 10 mm of Hg could be achieved in 81.8% of patients in supine position and 70% of patients in standing position. Thirteen patients responded to 5 mg dose and 9 patients required 10 mg. Postural hypotension and reflex tachycardia were absent. Three patients has mild leg cramps and constipation. No deleterious effects were observed on liver, kidney and hemopoetic function, or on E.C.G. Changes. Amlodipine given once daily is effective and safe, and is a useful addition to the existing armamentarium of antihypertensive drugs.

Regulación de la hematopoyesis / Hematopoyesis regulation

Las señales positivas y negativas juegan un papel trascendental en la regulación del sistema hematopoyético. En los últimos 30 años se purificaron más de 20 moléculas (glicoproteínas) con actividad biológica sobre las células progenitoras del sistema hematopoyético y sobre las células maduras circulantes. Los factores de crecimiento hematopoyéticos son las más conocidas de estas biomoléculas (citocinas como los factores estimulantes de colonias y las interleucinas), las cuales son capaces de estimular a las células de la médula ósea para producir descendencia madura. Hasta el presente se ha podido determianr no sólo la secuencia de la mayoría de estas glicoproteínas y los gene que las codifican, sino también caracterizar a las células blanco de cada uno de los factores y a sus receptores celulares. Los estudios de la interacción entre las células progenitoras hematopoyéticas y los factores que estimulan y/o inhiben su proliferación, han demostrado ser muy útiles en el desarrollo de terapias clínicas y podrían ser herramientas importantes en el análisis de la patogenia de muchas enfermedades hematológicas. Sin embargo, los mecanismos subyacentes en la producción de factores estimulantes o inhibidores y la proliferación de las células progenitoras hematopoyéticas continúan siendo escasamente conocidos

Tratamento de sindromes mielodisplásicos con danazol / Treatment of myelodysplastic syndromes with danazol

Se trataron con 600 mg de danazol, 42 pacientes con síndrome mielodisplásico (SMD) por un minimo de tres meses. En 10 pacientes se observó una buena respuesta, mientras que en 6 se observó una respuesta parcial: los restantes 26 pacientes fueron considerados como fracasos. La mediana de la supervivencia de los pacientes que respondieron fue de 43 meses, la cual es significativamente mejor que los 7 meses que se observan en los pacientes en que esta forma de tratamiento fue considerada como fracaso (p v 0.01). La toxidad observada fue mínima y no fue necesario modificar el programa terapéutico por esta razón. Los subgrupos morfológicos con anemia refractaria y con anemia refractaria con exceso de blastos fueron en los que su tuvieron en mayor número de respuestas positivas. El uso de danazol en pacientes con SMD debe ser considerado como una opción, ya que además de tener uan toxicidad mínima, puede modificar de manera significativa los parámetros hematológicos y la supervivencia. Este fármaco, podría considerarse para el tratamiento inicial, antes de que otros, generalmente más tóxicos, sea usado en los SMD

A quantitative method for evaluating the effects of drugs on bone marrow cellularity using the same experimental animal: quantification of the effects of a single dose busulphan

Se describe un método que permite cuantificar la celuradidad de la médula ósea de la rata por unidad de peso. A tal efecto se determinaron en diferentes tiempos los números absolutos de cada tipo celular presentes en el fémur derecho e isquierdo del mismo animal de experimentación y los resultados se expresan en número de células por mg de médula ósea. En la rata normal se demostró que el número absoluto de cada tipo celular por mg de médula ósea es muy similar en el fémur derecho e izquierdo. Esto ocurre cuando el estudio de la celularidad medular se realiza simultáneamente en ambos huesos, y también cuando se comparan los resultados del estudio cuantitativo del fémur izquierdo con los del fémur derecho del mismo animal con 10 y 20 días de intervalo. Con el objeto de ratificar la validez del método para el estudio de la acción de drogas sobre la hematopoyesis medular, se observaron, además, los efectos de una dosis de busulfán (20 mg/Kg, vía oral), administrada inmediatamente después de que se realizó el estudio cuantitativo de la celularidad de la médula ósea del fémur izquierdo de una rata normal. Los resultados se compararon con los obtenidos de la médula ósea del fémur derecho de los mismos animales, 10 días después. Se demostró una marcada y significativa disminución del total de células nucleadas por mg de médula ósea, luego de ese período de observación. Los efectos celulares del busulfán fueron particularmente intensos sobre la progenie mieloide de médula ósea, con una reducción del