Chronic Injury of Mice Bone Marrow Multipotent Hematopoietic Progenitor Cells Induced by Ionizing Radiation / 中国实验血液学杂志

OBJECTIVE@#To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation.@*METHODS@#Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed.@*RESULTS@#Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05).@*CONCLUSION@#Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.

Intervenciones psicológicas en cuidadores primarios de pacientes con trasplante de células hematopoyéticas: Revisión descriptiva / Psychological interventions on primary care healthcare providers of patients with hematopoietic cells transplantation: Descriptive revision / Intervenções psicológicas em cuidadores primários de pacientes com transplante de células hematopoiéticas: Revisão descritiva

Resumen Introducción: El ser cuidador primario informal de un paciente con indicación médica de trasplante de células progenitoras hematopoyéticas puede tener consecuencias negativas en su salud mental y calidad de vida. Objetivo: Describir las intervenciones psicológicas disponibles para el cuidador primario de pacientes sometidos a trasplante de células hematopoyéticas. Metodología: Se realizó una búsqueda sistematizada de los últimos 10 años con los términos MeSH: psychotherapy AND caregive AND stem cell transplantation en las principales bases de datos médicas y de psicología, para su análisis se empleó la estrategia: Problema, Intervención, Comparación y Outcomes (PICO). Resultados: Se identificaron 122 artículos, de ellos diez cumplieron los criterios de inclusión. Las intervenciones provenían de profesionales de enfermería o trabajo social; el 50% incluyó diadas (paciente y cuidador primario), mostraron una tendencia de duración corta, enfocada al periodo posterior al trasplante. Se basan en el entrenamiento en solución de problemas, manejo de estrés, atención plena y expresión emocional. Las intervenciones lograron la disminución de la depresión, ansiedad y estrés en el cuidador; pero no alcanzaron permanencia en la significancia estadística de dichos restablecimientos. Discusión: De acuerdo con lo observado en las publicaciones y por su impacto positivo en la salud mental, se recomienda la implementación de intervenciones psicológicas en cuidadores de pacientes con trasplante de células progenitoras hematopoyéticas. Conclusión: El apoyo psicológico brindado al cuidador generalmente es de profesionales de la salud que no pertenecen al área de la psicología, con resultados clínicos favorables en las etapas más críticas de su estado mental.

Abstract Introduction: Being an informal primary healthcare provider of a patient who undergoes hematopoietic progeny cells transplantation can have adverse consequences on mental health and the quality of life. Objective: To describe the available psychological interventions for the primary healthcare provider of patients undergoing hematopoietic cells transplantations. Methodology: A systematized search of the last 10 years using the MeSH terms psychotherapy AND caregiver AND stem cell transplantation was conducted on the main medical and psychological databases. The analysis strategy followed the PICO scheme (Problem, Intervention, Comparison, Outcomes). Results: 122 articles were identified, and 10 of them fulfilled the inclusion criteria. The interventions were related to nursing or social work professionals. 50% described patient-healthcare provider dyads with short interventions focused on the post-transplantation period. Discussion: According to what has been observed in the publications and due to its positive impact on mental health, the implementation of psychological interventions is recommended in caregivers of patients who underwent hematopoietic stem cell transplantation. Conclusion: The psychological support provided to the caregiver comes mainly from health professionals who do not belong to the area of psychology, with favorable clinical results in the most critical periods for their mental state.

Resumo Introdução: Ser cuidador primário informal de um paciente sometido a transplante de células progenitoras hematopoiéticas pode ter consequências negativas na saúde mental e na qualidade de vida. Objetivo: Descrever as intervenções psicológicas disponíveis para o cuidador primário de pacientes sometidos a transplante de células hematopoiéticas. Metodologia: Realizou-se uma busca sistematizada dos últimos 10 anos com os termos MeSH: psychotherapy AND caregive AND stem cell transplantation nas principais bases de dados médicas e de psicologia, para sua análise realizou-se a estratégia: Problema, Intervenção, Comparação e Outcomes (PICO). Resultados: Identificaram-se 122 artigos, dos quais, dez cumpriram os critérios de inclusão. As intervenções provinham de profissionais em enfermagem ou trabalho social; o 50% incluiu díades (paciente e cuidador primário), mostraram uma tendência de duração curta, focalizada no período posterior ao transplante. Baseiam-se no treinamento em solução de problemas, manejo de estresse, atenção plena e expressão emocional. As intervenções conseguiram melhoras clínicas na diminuição da depressão, ansiedade e estresse no cuidador; mas não alcançaram permanência na significância estatística destes restabelecimentos. Discussão: Conforme o observado nas publicações e por seu impacto positivo na saúde mental, recomenda-se a implementação de intervenções psicológicas em cuidadores de pacientes para quem se indicou transplante de células progenitoras hematopoiéticas. Conclusão: O apoio psicológico oferecido ao cuidador vem de principalmente profissionais da saúde que não pertencem à área da psicologia, com resultados clínicos favoráveis nos períodos mais críticos para seu estado mental.

CD34+ hematopoietic progenitor cells enumeration and mononuclear cell count: an experience from a tertiary care centre

Background: This study was conducted to assess the CD34+ hematopoietic progenitor cells enumeration by flow cytometry and the utility of performing mononuclear cell count before performing the Stem cell enumeration. Collection was done on two consecutive days with CD34+ hematopoietic progenitor cell enumeration of both the samples. Mononuclear cell counts were done in all the patients. The purpose of the study was to do counts directly from the leukapheresis pack and see the reliability of this practice.Methods: Samples were collected from the leukapheresis pack and subjected to mononuclear cell count and CD34+ hematopoietic progenitor cells enumeration by flow cytometty before harvesting.Results: A total of 66 samples from 34 patients were taken up for the study. 76.47% of our cases were that of multiple myeloma and 17.64% of the cases were that of non Hodgkin lymphoma and 2.94% cases each of neuroblastoma and Hodgkin lymphoma. It was noted that the mononuclear cell counts correlated well with the CD34+ HPC in most of the cases with MNC being above 4 x 108 per pack per kg body weight in cases where CD34+ HPC counts were more than the desired lower limit of 2 x 106 per pack per kg body weight.Conclusions: It was observed that flow cytometric enumeration of CD34+ hematopoietic progenitor cells directly from the leukapheresis pack gave satisfactory results even without doing peripheral blood CD34+ HPCs enumeration before leukapheresis. Also, in our study we were able to set a limit of mononuclear Cell at 4 x 108 per pack/kg BW as counts beyond that always correlated with the more accurate flow cytometric method of CD34+ HPC count of more than 2 x106 per pack/kg body weight, therefore acting as a crude method for assessing the mobilization.

Expression of adhesion molecules on CD34+ cells from steady-state bone marrow before and after mobilization and their association with the yield of CD34+ cells

BACKGROUND: Cell adhesion molecules (CAMs) expressed on hematopoietic progenitor cells (HPCs), endothelial cells, and stromal cells play a pivotal role in the mobilization of CD34+ cells. Herein, we conducted a non-randomized peripheral blood stem cell (PBSC) mobilization study aimed to compare the potential differences in the expressions of several CAMs and chemokines on CD34+ cells obtained from bone marrow aspirate before and after HPC mobilization from patients with hematologic malignancies and healthy donors. METHODS: Three-color cytofluorometric analysis was used to compare the expressions of CAMs and chemokines in the bone marrow before and after mobilization. RESULTS: For all studied groups, CAM expression among those with good and poor yields of CD34+ cells was significantly correlated with VCAM-1 (P=0.007), CD44 (P=0.027), and VLA-4 (P=0.014) expressions. VCAM-1 (P=0.001), FLT-3 (P=0.001), CD44 (P=0.011), VLA-4 (P=0.001), and LFA-1 (P=0.001) expressions were higher before HPC mobilization than after HPC mobilization. By contrast, the expression of CXCR4 significantly varied before and after mobilization only among those with successful PBSC mobilization (P=0.002). CONCLUSION: We attempted to identify particular aspects of CAMs involved in CD34+ cell mobilization, which is a highly complex mechanism that involves adhesion molecules and matrix metalloproteases. The mechanism by which CD34+ cell mobilization is activated through proteolytic enzymes is not fully understood. We believe that CXCR4, VLA-4, CD44, and VCAM-1 are the most important molecules implicated in HPC mobilization, particularly because they show a correlation with the yield of CD34+ cells collected via large volume leukapheresis.

The effect and mechanism of indole-3-carbinol on alleviating the radiation-induced bone marrow hematopoietic cell injury in mice / 天津医药

Objective To investigate the protective effect of indole-3-carbinol (I3C) on radiation-induced mouse bone marrow hematopoietic cell injury and the involved mechanisms. Methods (1) The bone marrow nuclear cells (BMNCs) from CD45.1 subtype of C57BL/6J mice were collected by a density gradient centrifugation method. The BMNCs were pretreated with a series doses of I3C (0 mol/L, 10-8 mol/L-10-3 mol/L) and then exposed with radiation of 137Csγ-ray (doses of irradiation were 0 Gy, 1 Gy and 4 Gy). After 18-hour culturing, the bioluminescence method was used to detect the cell viability. (2) These cells were divided into control group and 10-6 mol/L I3C group. Both groups were received the irradiation (0 Gy, 1 Gy and 4 Gy) and inoculated into the methylcellulose semi-solid culture medium to incubate 7 days, the colony forming unit-granulocyte monocytes (CFU-GM) were observed. (3) Twenty-four CD45.2 subtype mice used as the receptor were exposed with 8 Gy radiation. The CD45.1 BMNCs were divided into control group, 4 Gy irradiation group, 4 Gy irradiation and 10-6 mol/L I3C group. Donor cells were harvested from C57BL/6J (CD45.1) mice after they received various treatments, and were then mixed with competitive BMNCs from C57BL/6J (CD45.2) mice. The mixed cells were transplanted into recipient mice (8 mice/group). Flow cytometry was used to analyze the proportion of donor cells in peripheral blood of receptor. (4) The cells were divided into control group, 10-6 mol/L I3C group, 1 Gy irradiation group, 1 Gy irradiation with 10-6 mol/L I3C group. After 24-hour culturing, Western blot assay was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Results (1) I3C showed a significant cytotoxic effect on the BMNCs when its concentration was above 10-4 mol/L. 10-7-10-6 mol/L I3C could reduce the radiation injury of BMNCs under the same dose of irradiation. Therefore, 10-6 mol/L I3C was chosen for subsequent experiments. (2) The CFU-GM was significantly higher in 10-6 mol/L I3C group than that of control group (P<0.05). (3) Results of flow cytometry showed that the proportion of donor cells in receptor was significantly higher in 4 Gy irradiation group than that of control group, which decreased the engraftment capability of irradiated HSCs (P<0.05), although the engraftment capability of irradiated HSCs improved after 10-6 mol/L I3C treatment. (4) I3C significantly enhanced the increased protein expression of Nrf2 and HO-1 caused by radiation (P<0.05). Conclusion I3C has a protective effect on hematopoietic cells following radiation-induced injury, which may be related with activating the Nrf2/HO-1 signal pathway.

Influence of HPK1 overexpression in proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.

Influence of HPK1 overexpression in proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.

Hematopoietic progenitor cell mobilization for autologous transplantation - a literature review

ABSTRACT The use of high-dose chemotherapy with autologous support of hematopoietic progenitor cells is an effective strategy to treat various hematologic neoplasms, such as non-Hodgkin lymphomas and multiple myeloma. Mobilized peripheral blood progenitor cells are the main source of support for autologous transplants, and collection of an adequate number of hematopoietic progenitor cells is a critical step in the autologous transplant procedure. Traditional strategies, based on the use of growth factors with or without chemotherapy, have limitations even when remobilizations are performed. Granulocyte colony-stimulating factor is the most widely used agent for progenitor cell mobilization. The association of plerixafor, a C-X-C Chemokine receptor type 4 (CXCR4) inhibitor, to granulocyte colony stimulating factor generates rapid mobilization of hematopoietic progenitor cells. A literature review was performed of randomized studies comparing different mobilization schemes in the treatment of multiple myeloma and lymphomas to analyze their limitations and effectiveness in hematopoietic progenitor cell mobilization for autologous transplant. This analysis showed that the addition of plerixafor to granulocyte colony stimulating factor is well tolerated and results in a greater proportion of patients with non-Hodgkin lymphomas or multiple myeloma reaching optimal CD34+ cell collections with a smaller number of apheresis compared the use of granulocyte colony stimulating factor alone.

Effects of Astragalus-Angelica compatibility on proliferation of hematopoietic progenitor cells in mice with bone marrow hematopoiesis suppression / 中草药

Objective: To investigate the effects of different proportions of Astragalus and Angelica on the proliferation ability and cell senescence of hematopoietic progenitor cells (HPC) in the mice model of bone marrow hematopoiesis suppression, and to probe the mechanism of Astragalus-Angelica compatibility on promoting hematopoiesis. Methods: ICR male mice were randomly divided into normal group, model group, positive control group of recombinant human granulocyte colony stimulating factor (rhG-CSF), Astragalus group, Angelica group, and different proportion combination groups of Astragalus and Angelica, and the animals in Chinese medicinal herb groups were ig administered, once a day, for 8 d. In the positive control group, rhG-CSF was sc injected on days 6, 7, and 8 of administration. Except for the normal group, the others were received cyclophosphamide (CTX) by ip injection on days 4, 5, and 6 of administration to establish the model of bone marrow hemopoiesis suppression. The mice were killed on day 9 to obtain bone marrow cells for the culture of HPC, and the senescence rate of bone marrow nucleated cell (BMNC) was detected by SA-β-galactosidase staining method. Results: Compared with the model group, Angelica and Astragalus-Angelica with proportions of 5:1, 1:1, and 1:5 could significantly raise colony forming unit-granulocyte (CFU-GM), colony forming unit-megakaryocyte (CFU-MK), colony forming unit-erythroid (CFU-E), and burst forming unit-erythroid (BFU-E) (P < 0.01). Astragalus-Angelica with 10:1 made CFU-MK, CFU-E, BFU-E remarkably increase (P < 0.05, 0.01), and had no effect on CFU-GM. Furthermore, Astragalus-Angelica with 1:1 promoting the formation of HPC was evidently stronger than that of single Astragalus, single Angelica, and other combinations (P < 0.05). Compared with the model group, the senescence positive rate of BMNC was markedly decreased in Astragalus group, Angelica group, and Astragalus-Angelica combination groups (P < 0.01), while being lowest in Astragalus-Angelica with 1:1 proportion (P < 0.05, 0.01). Conclusion: Astragalus, Angelica, Astragalus-Angelica with 10:1, 5:1, 1:1, and 1:5 proportions can prompt the proliferation and differentiation of HPC that bone marrow hematopoiesis is suppressed in mice, inhibit the senescence of bone marrow hematopoietic cell, moreover, the effect of Astragalus-Angelica with 1:1 proportion is the best. It suggests that promoting the proliferation and differentiation of HPC is one of the mechanisms that Astragalus-Angelica prompt hematopoiesis.

The difference in the radiosensitivity between hematopoietic stem and progenitor cells after different doses of 137Csγ-radiation / 天津医药

Objective To observe the different radiosensitivity induced by different doses of 137Csγ-ray irradiation between hematopoietic stem and progenitor cells. Methods Seventy-two C57BL/6 mice were randomly divided into control group and irradiated groups (2, 4 and 6 Csγ-ray irradiation, n=18 for each group). Mice of control group received sham irradiation, and the rest accepted 2, 4 and 6 Gy137Csγtotal body irradiation, respectively. After 14-day, 35-day and 56-day irradiation, the peripheral blood samples were collected by balls enucleation. The number of bone marrow nuclear cells, hematopoietic stem and progenitor cells were counted. Results The peripheral blood of irradiated mice showed significant changes in the number of white blood cells (WBC), red blood cells (RBC), platelets (PLT) and hemoglobin (HGB) in a dose-response relationship. Compared with the control group, the numbers of BMNCs and hematopoietic progenitor cells (HPCs) were significantly lower in irradiated group. At 35 d and 56 d after 6 Gy irradiation the numbers of BMNCs and HPCs were significantly lower than those of control group (P<0.05). There were no significant differences in numbers of BMNCs and HPCs between irradiated groups (2 and 4 Gy) and control group. The number of bone marrow hematopoietic stem cells (HSCs) was significantly lower in irradiated group than that in control group after 14-d and 56-d irradiation (P<0.05). Conclusion 137Csγ-ray irradiation has some damage in mouse hematopoietic system. The damage caused by radiation is persistent to hematopoietic stem cells.

Effect of different doses of HS6101 on hematopoietic function of ICR mice treated with cyclophosphamide / 国际药学研究杂志

Objective: To observe the effect of different doses of HS6101 on the recovery of hematopoietic injury in ICR mice treated with cyclophosphamideCTX. Methods: Normal ICR mice were intraperitoneally injected with CTX at 100 mg/kg once a day for 3 consecutive days， and the mouse model of chemotherapy-induced hematopoietic injury was established. Three groups of mice with 20 per group， were respectively injected with HS6101 at 0， 9 or 27 μg subcutaneously at one hour before the first administration of CTX. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated at 4 d and 9 d after the first administration of CTX. Results: In ICR mice after chemotherapy with CTX， all doses of HS6101 significantly increased peripheral leukocytes and neutrophils P<0.05， elevated the number of multilineage hematopoietic progenitor cell colonies of bone marrow， and stimulated the proliferation of bone marrow cells after CTX injury. Mice receiving 27 μg HS6101 were better than those of the other two groups. Conclusion: HS6101 at 27 μg could significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy.

Effect of different doses of HS6101 on hematopoietic function of ICR mice treated with cyclophosphamide / 国际药学研究杂志

Objective To observe the effect of different doses of HS6101 on the recovery of hematopoietic injury in ICR mice treated with cyclophosphamide (CTX). Methods Normal ICR mice were intraperitoneally injected with CTX at 100 mg/kg once a day for 3 consecutive days,and the mouse model of chemotherapy-induced hematopoietic injury was established. Three groups of mice (with 20 per group),were respectively injected with HS6101 at 0,9 or 27 μg subcutaneously at one hour before the first administration of CTX. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated at 4 d and 9 d after the first administration of CTX. Results In ICR mice after chemotherapy with CTX,all doses of HS6101 significantly increased peripheral leukocytes and neutrophils (P<0.05),elevated the number of multilineage hematopoietic progenitor cell colonies of bone marrow,and stimulated the proliferation of bone marrow cells after CTX injury. Mice receiving 27 μg HS6101 were better than those of the other two groups. Conclusion HS6101 at 27 μg could significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy.

Effect of Bu-Shen Yi-Sui Sheng-Xue Method for Medicated Serum of Rats with Aplastic Anemia on Mechanism of Proliferation and Differentiation of Hematopoietic Progenitor Cells / 世界科学技术-中医药现代化

This study was aimed to investigate the effect of Bu-Shen Y i-Sui Sheng-Xue (BSYSSX) method on pro-liferation and differentiation mechanisms of hematopoietic progenitor cells. The rat models were established by 60Co-γrays and cyclophosphamide. Compound Chinese medicine was gavaged to rats of the normal control group, model group, stanozolol group, Yi-Sui Sheng-Xue (YSSX) group, Wen-Shen Sheng-Xue(WSSX) group and Zi-Shen Sheng-Xue (ZSSX) group. Then, serum of rat was prepared. Rat bone marrow cells were incubated with AA rats serum ac-counted for 20% and the number of hematopoietic progenitor cells colony-forming units (CFU) were counted. The level of GATA-1 and PU.1 mRNA in colony cells were detected with RT-PCR. The results showed that compared with the normal control group, the number of bone marrow cells, CFU-E, BFU-E, CFU-GM, as well as the expres-sion of GATA-1 and PU.1 mRNA in the model group decreased significantly (P< 0.01). Compared with the model group, the number of bone marrow cells, CFU-E, BFU-E, CFU-GM of each treatment group were significantly in-creased (P< 0.01). CFU-E and BFU-E of the ZSSX group were better than the YSSX group (P < 0.01). CFU-GM of the ZSSX group was better than the YSSX group and the WSSX group. The expression of GATA-1 and PU.1 mR-NA in each treatment group were significantly higher than the model group (P< 0.01). The expression of GATA-1 mRNA in the ZSSX group was better than the YSSZ group and WSSX group (P< 0.05). The expression of PU.1 mR-NA in the ZSSX group was higher than the YSSX group and WSSX group. It was concluded that BSYSSX method may increase the expression of GATA-1 and PU.1 mRNA in order to promote the proliferation and differentiation of bone marrow hematopoietic progenitor cells. The ZSSX method was better than the YSSX method and WSSX method.

Células progenitoras hematopoyéticas de sangre de cordón umbilical / Hematopoietic progenitor cells from umbilical cord blood

La sangre de cordón umbilical (SCU) es una rica fuente de células progenitoras hematopoyéticas (CPH) para el trasplante. Se describe la historia del uso de la SCU para trasplante hematopoyético y la necesidad del consentimiento informado para su obtención, conservación, almacenamiento y uso con estos fines. Se hace referencia a la seguridad para el receptor en cuanto a selección, requisitos epidemiológicos, obstétricos y analíticos de la muestra, la calidad hematopoyética de la colecta y procesamiento hasta su conservación, así como a las ventajas y desventajas de su uso en el trasplante. Por último, se aborda la importancia de incrementar en los bancos de SCU, el almacenamiento de CPH que tengan representación antigénica compatible con las minorías étnicas

The umbilical cord blood (UCB) is a rich source of hematopoietic progenitor cells (HPC) for transplantation. Here, we describe the history of using of UCB for hematopoietic transplantation and the need of obtaining informed consent for its conservation, storage and further use for these purposes. Reference is made to the security for the receiver in terms of sample selection, epidemiological, obstetrical, and analytical requirements, as well as the quality of hematopoietic collection and processing for preservation, in addition to the advantages and disadvantages of its use in transplantation. Finally, we address the importance of increasing the UCB banks, storing CPH with antigenic representation compatible with ethnic minorities

A infecção das células progenitoras hematopoiéticas pelo HIV e a relação com a persistência viral e anormalidades hematológicas / Infection of hematopoietic progenitor cells by HIV and the relation with viral persistence and hematological abnormalities

O vírus da imunodeficiência humana causa infecção persistente apesar da terapia com antirretrovirais. Isso leva a crer que existem outros reservatórios virais além dos já conhecidos. Estudos recentes mostram que o vírus pode invadir as células progenitoras hematopoiéticas, causar infecção ativa assim como latente e morte celular. Essas célulasinfectadas latentemente podem persistir por vários anos e contribuir para a viremia residual e a persistência viral. A morte celular causada pela invasão e replicação do vírus pode estar associada a anormalidades hematopoiéticas. Palavras-chave. Células progenitoras hematopoiéticas; vírus da imunodeficiência adquirida; reservatórios virais

Human immunodeficiency virus (HIV) causes persistent infection despite antiretroviral therapy. This suggests the existence of other viral reservoirs beyond those already known. Recent studies show that HIV can invade hematopoietic progenitor cells, generating active infection as well as latent infection and cell death. The latently infected HPCs may persist for several years and contribute to residual viremia and viral persistence. Cell death caused by the invasion and replication of the virus may be associated with hematopoietic abnormalities.

Evaluación de la contaminación microbiológica en productos de células progenitoras hematopoyéticas. Experiencia de un laboratorio de procesamiento de CPH y Banco de Sangre de Cordón Umbilical / Microbiological evaluation in hematopoietic progenitor cell products. Experience from an HPC processing laboratory and Cord Blood Bank

Los productos de células progenitoras hematopoyéticas (CPH) representan una fuente potencial de infección en pacientes inmunosuprimidos que reciben infusión de CPH como parte de su tratamiento. La probabilidad de contaminación de cada producto difiere según la técnica de colecta y el procesamiento aplicado. En este trabajo hemos realizado un análisis retrospectivo de los resultados de cultivos microbiológicos de 1707 productos de CPH obtenidos de sus tres fuentes (médula ósea, sangre periférica y sangre de cordón umbilical) con el objetivo de determinar la proporción de unidades contaminadas. Además, fueron comparadas las distintas técnicas de colecta y las diferentes manipulaciones a las que fueron sometidos los productos. Por otro lado, se analizaron las posibles fuentes de contaminación según el microorganismo identificado y se evaluó la supervivencia de los mismos luego del descongelamiento. La prevalencia de productos de CPH con cultivos microbiológicos positivos reportados en este estudio (5,2%) se corresponde con lo descripto en la literatura. No encontramos diferencias significativas al comparar los productos según la fuente de la cual provenían las CPH. Tampoco hubo diferencias según los procedimientos aplicados a cada unidad. Los microorganismos aislados en los productos de CPH fueron los esperados de acuerdo a la fuente de la cual provenían las células. Pudo comprobarse que algunos de ellos son capaces de sobrevivir a los procesos de criopreservación y descongelamiento. La estricta adhesión a las normas de buenas prácticas de manufactura y buenas prácticas tisulares es un requisito para minimizar los riesgos de introducir microorganismos contaminantes. Disponer de un producto de CPH seguro es fundamental para el éxito de un trasplante.

Hematopoietic stem cell products (HSCP) represent a potential source of infection for immunosuprressed patients that receive HSCP infusion as part of their treatment. The probability of contamination of a HSCP product depends on the collection technique as well as the processing performed. In this work we have carried out a retrospective analysis of the results of 1707 microbiological cultures of HSCP products obtained from three different sources: bone marrow, peripheral blood and umbilical cord blood. We determined the proportion of HSCP units that were contaminated by microorganisms. Furthermore we compared the results obtained with different collection techniques and with the distinct manipulations that were used for processing. Moreover we analysed the possible sources of contamination related to the microorganisms identified and we evaluated the survival of them after thawing. The proportion of HSCP products with positive microbiological cultures obtained in this study (5,2%) correlates with that reported by other authors. We have not found significant differences between the results achieved with HSCP products from different sources. There were neither differences depending on the procedures applied. The isolated microorganisms from the HSCP products were the expected in accordance with the source of the cells. It could be demonstrated that some of them were capable of surviving the cryopreservation and thawing processes. Adherence to good manufacture practices and good tissue practices regulations is critical for minimizing the risks of introducing contaminant microorganisms. A safe HSCP product is essential for the success of a transplant.

Leucemia Mielóide Crônica: transplante de medula óssea / Cronic Myeloid Leukemia: bone marrow transplantation

A única terapia curativa para Leucemia Mielóide Crônica ainda é o transplante de células hematopoéticas progenitoras (TCHP); os atuais resultados com uso do imatinibe são suficientes para indicarmos o TCHP como um tratamento de segunda ou terceira linha. A decisão de realizar TCHP existe em uma variedade de momentos: falha em se conseguir remissão hemtológica, citogenética ou molecular, ou quando se perde a melhor resposta conseguida ou por progressão da doença para uma fase avançada. Há decisão também de como transplantar. Nesta revisão apontamos algumas destas decisões e as atuais controvérsias.

Although the only curative therapy for chronic myeloid leukemia remains allogeneic stem cell transplantation (allo-SCT), the results of imatinib in newly diagnosed patients are sufficiently impressive to have displaced allo-SCT to second or third-line treatment. Patients now arrive at a decision for transplantation in a variety of disease situations: failing to achieve certain hematological, cytogenetic and molecular remission by some pre-determined timepoint, having lost a previous best response or due to progression to an advanced phase. The decision is also how to transplant. In this review article, the evidence supporting some of these decisions and current controversies are discussed.

Avaliação do valor prognóstico da detecção do status quimérico de pacientes após o transplante alogênico de células progenitoras hematopoéticas / Evaluation of prognostic importance of the detection of chimerical status in post-allogeneic progenitor cell transplantation patients

Este trabalho teve por objetivo correlacionar o status quimérico de pacientes pós -TCPH alogênico com parâmetros clínicos, para avaliar o valor preditivo dos achados laboratorias de quimerismo. Amostras de sangue de 98 pacientes (67 em seguimento e 31 novos casos) foram submetidas à análise do status quimérico pós-TCPH. Os "loci"analisados por biologia molecular foram CS1PO, TPOX, F13A1, FESFPS, HUMTH01, VWA, SE33, HUMARA, HUMD21S11 e Amelogenina. Precocidade da evidência laboratorial de quimerismo misto (QM), em relação ao aparecimento dos sintomas clínicos de recaída, foi observada em 9 dos 12 pacientes nas LA, ou seja, nesses casos, a primeira manifestação de QM foi detectada pelo exame laboratorial antes de qualquer evidência citológica ou clínica de recaída. Em todos eles, houve uma mudança terapêutica relacionada com esse momento do aparecimento do QM. Em 100 por cento dos pacientes com QM na LMC, a detecção do quimerismo pelo exame laboratorial foi anterior a qualquer evidência citológica ou clínica de recaída. De uma maneira geral, o exame laboratorial da avaliação do status quimérico pós-TCPH alogênico pela análise dos "loci"hipervariáveis do genoma, mostrou ser um exame sensível, com detecção de até 1 por cento de QM e precoce, visto que, muitas vezes, foi a primeira manifestação de doença residual antes de qualquer evidência citológica ou clínica da mesma. A associação da existência de QM e a recaída clínica e/ou óbito fica mais evidente nos casos de LA do que nos casos de LMC e AAS.

This study aimed to correlate the chimerical status in post-allogeneic hematopoietic stem cell transplantation (HSCT) patients to clinical patterns in order to evaluate the predictive value of chimerism laboratorial findings. Blood samples from 98 patients (67 current and 31 new cases) were submitted to post-HSCT chimerical status analysis. The CS1PO, TPOX, F13A1, FESFPS, HUMTH01, VWA, SE33, HUMARA, HUMD21S11 and Amelogenian loci were analyzed. Precocity of Mixed Chimerism (MC) laboratorial evidence in relation to recurrent clinical symptom manifestations was observed in 9 out of 12 patients in AL, i.e., in these cases the first MC manifestation was detected in laboratory tests before any cytological or clinical evidence. In all cases, there was a therapeutic change due to MC onset. Chimerism detection through laboratorial examinations was prior to any cytological or clinical evidence in 100 percent of patients presenting MC in CML. Considering that it was the first manifestation of residual disease, before any cytological or clinical manifestation, laboratorial examination to evaluate chimerical status in post-allogeneic hematopoietic stem cell transplantation through analysis of genome hyper-variable loci, turned out to be a more sensitive examination and presented a detection rate of up to 1 percent for early MC. The association of MC to clinical recurrence and/death is more evident in AL cases than in CML and SAA.

Inverse Tendency between Ex Vivo Expansion Potential of Hematopoietic Progenitors and Time to Engraftment after Hematopoietic Stem Cell Transplantation / 대한진단검사의학회지

BACKGROUND: The CD34+ cell dose and infused number of committed progenitor cells in transplantation are important factors in hematologic engraftment. However, the relationship between expansion potential of progenitor cells and hematologic engraftment remains controversial. We evaluated whether expansion potential of progenitor cells is a predictive factor of post-transplantation hematologic engraftment. METHODS: Mononuclear cells isolated from mobilized peripheral blood and bone marrow were cultured with cytokine cocktail for 7 days. Progenitor cells and committed progenitors were analyzed using stem cell markers (CD34 and CD133) and lineage specific markers. Hematologic engraftment was defined as neutrophil counts over 500/microliter and platelet counts over 20,000/microliter without transfusion. Acute and chronic graft-versus-host disease (GVHD) were investigated. RESULTS: There was inverse tendency between the number and fold expansion of progenitor cells or committed (granulocytic or megakaryocytic) progenitors and time to engraftment. Especially, fold expansion of CD34(+)/CD33(+) cells was significantly correlated with time to neutrophil engraftment in bone marrow transplantation (r=-0.56, P=0.04). The infused number and fold expansion of lymphoid progenitors were not related to the occurrence of acute or chronic GVHD. CONCLUSIONS: We could not prove that expansion potential of progenitor cells and committed progenitor cells is correlated to hematologic engraftment although there is a correlation between CD34(+)/ CD33(+) cells and time to neutrophil engraftment. But, a further study on the value of expansion potential is required because there is an inverse tendency.

Alteraciones renales en el trasplante de células progenitoras hematopoyéticas / Renal alterations in hematopoietic progenitor cell transplantation

Se realizó un estudio retrospectivo y descriptivo de 97 pacientes sometidos a trasplante de células progenitoras hematopoyéticas (TCPH), entre los años 1985 y 2004, para evaluar sus alteraciones renales. Se observó una incidencia de esta complicación del 27,5 % en la muestra analizada. Las alteraciones renales más frecuentes resultaron la disfunción y la insuficiencia renales agudas (51,6 y 22,5 %, respectivamente). Durante los primeros 120 d pos -TCPH predominó la disfunción renal aguda y en el período de más de 120 d, la nefritis radiógena. Predominaron como causas la multifactorial (54,2 %) y la nefrotoxicidad por ciclosporina A (17,1 %): hasta los 30 d, la multifactorial (72,7 %); entre los 31 y 120, la nefrotoxicidad por ciclosporina A (71,4 %) y en el mayor de 120, las radiaciones (50,0 %). Se observaron más alteraciones renales entre los trasplantados alogénicos (61,5 %) que entre los autólogos (15,4 %) y en los sometidos a QMT+RT como régimen condicionante (30,8 %), que los que recibieron solo QMT (11,7 %). La aplicación en el régimen condicionante de las dosis únicas de radioterapia se correlacionó con la aparición de la nefritis radiógena.

A retrospective and descriptive study was conducted among 97 patients that underwent hematopoietic progenitor cell transplantation (HPCT) between 1985 and 2004 to evaluate their renal alterations. An incidence of this complication of 27.5 % was observed in the analysed sample. The most frequent renal alterations were acute renal dysfunction and failure. (51.6 and 22.5 %, respectively). During the first 120 days of the transplantation, there was a predominance of acute renal dysfunction, whereas in the period of more than 120 days there was a prevalence of radiogenic nephritis. The prevailing causes were the multifactorial (54.2 %) and nephrotoxicity due to cyclosporin A (17.1 %): up to 30 days, the multifactorial (72.7 %); between 31 and 120 days, the nephrotoxicity due to cyclosporin A (71.4 %); and in the period over 120 days, the radiations (50 %). More alterations were observed in the allogenous transplant recipients (61.5 %) than in the autologous transplant recipients (15.4 %). Among those who underwent CMT + RT as a conditioning regime (30.8 %) there were also more alterations than among those who received only CMT (11.7 %). The application in the conditioning regime of the unique doses of radiotherapy was correlated with the appearance of radiogenic nephritis.