OBJECTIVETo explore the formation of pre-metastatic niche in the mouse lung and to study the underlying molecular mechanisms whereby primary breast carcinoma-derived factors mediate recruitment of bone marrow-derived cells (BMDCs) and affect the formation of pre-metastatic lung environment before the arrival of tumor cells.

METHODSMammary carcinoma 4T1 cells were inoculated into the mammary gland to construct mouse model of breast cancer. Confocal microscopy was used to detect the recruitment of BMDCs in the pre-metastatic lungs. The expression of factors in the mouse sera and 4T1 cell culture media was assayed using RayBio Custom mouse cytokine antibody array kit. The mice were injected daily with recombinant VEGF for 7 consecutive days to observe the effect of VEGF on BMDCs recruitment in the mouse lung.

RESULTSNo BMDCs were observed in the lungs of control and 4T1-tumor-bearing mice on day 0. On day 7 and 14, clusters of BMDCs observed in the lungs of 4T1-tumor-bearing mice were 8.7±2.2/objective field and 48.8±3.2/objective field, respectively, significantly higher than those in the control mice (1.1±0.8/objective field and 3.1±1.7/objective field) (P<0.05 for both). Confocal microscopic observation found that metastatic breast cancer cells preferentially facilitate BMDCs recruitment sites in the pre-metastatic mouse lungs. The levels of VEGF, GM-CSF, and IL-6 in the serum of 4T1-tumor-bearing mice were significantly increased compared with those in the control group (P<0.05 for all). However, VEGF was detected only in the culture media of 4T1 cells. The amount of BMDCs in the mouse lung tissue was (22.8±3.6)/objective field in the VEGF group and (3.1±0.4)/objective field in the control group (P<0.05). There were 36.8±5.4 metastatic foci in the lung tissue of VEGF group and 12.6±2.2 in the control group (P<0.05).

CONCLUSIONSThe results of this study demonstrate that primary breast cancer cells can alter the lung microenvironment during the pre-metastatic phase and induce the formation of pre-metastatic niche. Primary tumor cell-derived VEGF may be a crucial factor responsible for the formation of pre-metastatic niche.

Animals ; Bone Marrow Cells ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; Humans ; Interleukin-6 ; blood ; Lung ; pathology ; Lung Neoplasms ; secondary ; Mice ; Recombinant Proteins ; administration & dosage ; Time Factors ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A ; administration & dosage ; physiology ; secretion

Colchicine poisoning is rare but can cause potentially life-threatening toxic complications such as hypovolemic shock, cardiovascular collapse and multiple organ failure. In this case report, we describe a case of a 20-year-old female who presented to the emergency department after suicidal ingestion of a toxic dose of colchicine. She developed thrombocytopenia, neutropenia and acute respiratory distress syndrome that required blood transfusion and administration of granulocyte colony stimulating factor for the prevention of infectious complications. With regard to the clinical manifestations of colchicine toxicity, we discussed suggested mechanisms.
Blood Transfusion ; Colchicine* ; Colony-Stimulating Factors* ; Eating ; Emergency Service, Hospital ; Female ; Granulocytes* ; Humans ; Multiple Organ Failure ; Neutropenia ; Poisoning* ; Respiratory Distress Syndrome, Adult ; Shock ; Thrombocytopenia ; Young Adult

Colchicine poisoning is rare but can cause potentially life-threatening toxic complications such as hypovolemic shock, cardiovascular collapse and multiple organ failure. In this case report, we describe a case of a 20-year-old female who presented to the emergency department after suicidal ingestion of a toxic dose of colchicine. She developed thrombocytopenia, neutropenia and acute respiratory distress syndrome that required blood transfusion and administration of granulocyte colony stimulating factor for the prevention of infectious complications. With regard to the clinical manifestations of colchicine toxicity, we discussed suggested mechanisms.
Blood Transfusion ; Colchicine ; Colony-Stimulating Factors ; Eating ; Emergency Service, Hospital ; Female ; Granulocytes ; Humans ; Multiple Organ Failure ; Neutropenia ; Poisoning ; Respiratory Distress Syndrome, Adult ; Shock ; Thrombocytopenia ; Young Adult

BACKGROUND: Peripheral blood stem cell (PBSC) transplantation is a curative treatment in various hematologic malignancies and some solid cancers. Effective mobilization and collection of PBSC is essential for successful PBSC transplantation. The aim of this study was to investigate the useful factors for predicting PBSC collection using multivariate analysis. METHODS: We retrospectively reviewed the medical records of 170 allogeneic and 389 autologous donors at Chonnam National University Hwasun Hospital between 2005 and 2012. Donor groups were divided into three groups (failure group, suboptimal group, and optimal group) according to the total CD34+ yield. Donors were compared regarding age, sex, body weight, disease, complete blood count, hematopoietic progenitor cell (HPC) parameter of automated cell counter, process volume, number of leukapheresis procedures, prior mobilization history, type of vascular access and instrument. RESULTS: In allogeneic PBSC collections (n=170), the collection failure group showed lower baseline (premobilization) white blood cell (WBC) (P=0.004) and HPC (P<0.001) than the optimal group. In autologous PBSC collections (n=389), the collection failure group showed lower baseline HPC and more frequent prior mobilization history (P<0.001) than the suboptimal and optimal group. In multivariate analysis, older age, lower number of leukapheresis procedures, and prior mobilization history were risk factors associated with mobilization failure. CONCLUSION: Our data suggest that baseline WBC and HPC would be useful for predicting poor mobilizer in allogeneic PBSC collection, whereas baseline HPC would be useful in autologous PBSC collection. Conventional chemotherapy and G-CSF based remobilization would not be helpful to proven poor mobilizer in previous mobilization.
Blood Cell Count ; Body Weight ; Cell Count ; Drug Therapy ; Granulocyte Colony-Stimulating Factor ; Hematologic Neoplasms ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; Humans ; Jeollanam-do ; Leukapheresis ; Leukocytes ; Medical Records ; Multivariate Analysis ; Retrospective Studies ; Risk Factors ; Stem Cells* ; Tissue Donors ; Transplantation

BACKGROUND: Major burn injury induces an inflammatory response that is accompanied by the release of various cytokines. We investigated the gradual changes in the levels of pro-inflammatory and anti-inflammatory cytokines following burn injury and determined the relationship between these levels and burn size in adult Korean patients with burn injury. METHODS: Blood samples from 9 healthy controls and 60 Korean burn patients were collected on days 1, 3, 7, 14, and 21 after burn injury, and concentrations of interleukin (IL)-6, IL-8, IL-10, tumor necrosis factor (TNF)-alpha, and granulocyte-colony stimulating factor (G-CSF) were measured. Burn patients were divided into 3 groups according to burn size (15-30%, 31-50%, >50% total body surface area), and the concentrations of the cytokines were compared between these groups and the control group over 3 weeks. RESULTS: Compared to their levels in controls, IL-6, IL-8, IL-10, TNF-alpha, and G-CSF levels in burn patients were significantly higher during the observation period. Median concentrations of IL-8, IL-10, and G-CSF at each time point increased with burn size, although peak levels and time to peak levels of these cytokines differed from patient to patient. CONCLUSIONS: These findings indicate that IL-6, IL-8, IL-10, TNF-alpha, and G-CSF are important mediators in inflammatory changes after burn injury; however, various factors, including burn size, may influence the concentrations of these cytokines.
Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; Burns/blood/*pathology ; Granulocyte Colony-Stimulating Factor/*blood ; Humans ; Interleukin-10/*blood ; Interleukin-6/*blood ; Interleukin-8/*blood ; Male ; Middle Aged ; Republic of Korea ; Time Factors ; Tumor Necrosis Factor-alpha/*blood ; Young Adult

BACKGROUND: Major burn injury induces an inflammatory response that is accompanied by the release of various cytokines. We investigated the gradual changes in the levels of pro-inflammatory and anti-inflammatory cytokines following burn injury and determined the relationship between these levels and burn size in adult Korean patients with burn injury. METHODS: Blood samples from 9 healthy controls and 60 Korean burn patients were collected on days 1, 3, 7, 14, and 21 after burn injury, and concentrations of interleukin (IL)-6, IL-8, IL-10, tumor necrosis factor (TNF)-alpha, and granulocyte-colony stimulating factor (G-CSF) were measured. Burn patients were divided into 3 groups according to burn size (15-30%, 31-50%, >50% total body surface area), and the concentrations of the cytokines were compared between these groups and the control group over 3 weeks. RESULTS: Compared to their levels in controls, IL-6, IL-8, IL-10, TNF-alpha, and G-CSF levels in burn patients were significantly higher during the observation period. Median concentrations of IL-8, IL-10, and G-CSF at each time point increased with burn size, although peak levels and time to peak levels of these cytokines differed from patient to patient. CONCLUSIONS: These findings indicate that IL-6, IL-8, IL-10, TNF-alpha, and G-CSF are important mediators in inflammatory changes after burn injury; however, various factors, including burn size, may influence the concentrations of these cytokines.
Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; Burns/blood/*pathology ; Granulocyte Colony-Stimulating Factor/*blood ; Humans ; Interleukin-10/*blood ; Interleukin-6/*blood ; Interleukin-8/*blood ; Male ; Middle Aged ; Republic of Korea ; Time Factors ; Tumor Necrosis Factor-alpha/*blood ; Young Adult

In our study, a two-phase culture system was developed to acquire large amount of CD41+ and polyploidy cells. Human mobilized peripheral blood CD34+ (PB CD34+) cells were first cultured in expansion medium (Cocktail or CC100 medium) for 3,4,5 or 6 days, and then cultured in megakaryocytic differentiation medium containing TPO and SCF for additional 7, 8 or 9 days. Cell expansion, morphology, CD41+ cell percentage and DNA content were investigated to evaluate the protocol. The result showed that more CD41+ and polyploidy cells could be obtained following the two-phase culture with Cocktail medium than with CC100. Moreover, with 3 days expansion in Cocktail medium plus 7 days in differentiation medium, the initial CD 34+ cells obtained 16-fold expansion of CD41+ cells and 3-fold expansion of polyploidy cells, such obtained level being significantly higher than that of culturing cells with only one step in TPO or TPO+SCF. We conclude that with the two-phase culture system, PB CD34+ cells can expand and differentiate to more CD41+ and polyploidy cells than those cultured only in accordance to the one-stage culture protocol, so a new and highly efficient megakaryocyte differentiation model for megakaryocyte and platelet related researches is provided already.
Antigens, CD34 ; blood ; Blood Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; physiology ; Colony-Stimulating Factors ; physiology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Megakaryocytes ; cytology ; Stem Cells ; cytology

OBJECTIVETo investigate the optimal time for second allogeneic peripheral blood stem cell grafts (PBSC) harvest from healthy donors after in vivo recombinant human granulocyte colony-stimulating factor application (rhG-CSF).

METHODSThirty-eight healthy donors of second collection (group A) were treated with subcutaneous rhG-CSF \[5 microgxkg(-1)xd(-1)\] for five consecutive days and followed by leukapheresis on day 5 and 6. The control group (group B) was thirty-eight healthy donors who had received a first PBSC collection previously. Group A was reclassified as group C (< or = 9 months) and group D (> 9 months) according to the 75% quantile of interim time between first and second collection. The quantities of lymphocytes of CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry.

RESULTSThe median number of CD3(+)CD8(+) (25.51 x 10(8)) and CD34(+) cells (0.51 x 10(8)) in group A were significantly lower than that (31.55 x 10(8) and 0.70 x 10(8) respectively) in group B (P < 0.05), and so did the CD3(+)CD8(+) (23.42 x 10(8)) and CD34(+) cells (0.42 x 10(8)) in group C than that in group B (P < 0.05). There was no statistical difference in median numbers of T cell subsets, monocytes, and CD34(+) cells between group B and group D (P > 0.05). The cell ratios of CD4(+)/CD8(+), CD14(+)/CD3(+) and CD3(+)CD4(-)CD8(-) T/CD3(+) in PBSC in group A, group C, and group D were similar to that in group B (P > 0.05). Sperman analysis showed a positive correlation between the total CD34(+) cells in second collection and the interval time from first to second collection (r = 0.357, P = 0.028).

CONCLUSIONNine months after the first collection maybe an optimal time for the second PBSC collection. For those who undergo second PBSC collection within 9 months, more circulation blood should be extracted to ensure enough immunological and hematopoietic compositions.

Adolescent ; Adult ; Cytapheresis ; methods ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Time Factors ; Tissue Donors ; Transplantation, Homologous ; Young Adult

This study was aimed to explore the effect of donor characteristics (age, sex and so on.) on CD34(+) cell yields in apheresis from healthy donors mobilized by recombinant granulocyte colony-stimulating factor(rhG-CSF). In 61 healthy donors, the characteristics associated with CD34(+) cell yield were analysed. The relationship between the CD34(+) cell yields and donor characteristics was statistically assessed with multivariate forward, backward and stepwise regression methods. A variety of parameters were analyzed which included donor age, sex, weight, height, body mass index (BMI) and time for collection of peripheral blood apheresis, while the mean number of peripheral blood mononuclear cells (MNCs), CD34(+) cell count, CD34(+) cell proportion based on MNC and CD34(+) cell count per kg of donor weight were used as the variables. The results showed that age of donors was the main factor impacting CD34(+) cell yields (-0.60 < r < -0.45, p < 0.005). In a partial correlation analysis the body weight, height and BMI were served as control factors, the negative correlation of age with CD34(+) cell yields was still found (-0.50 < r < -0.35, p < 0.02). BMI was only weakly correlated with the yields of CD34(+) cells per kg(r = -0.297, p < 0.05). As a whole, sex showed no relation with the CD34(+) cell yields. Compared with the female group less than 35 years old, height, weight and BMI in male group of low age exerted a positive impact on CD34(+) cell yield. The optimal time for collection of PB was day 4 after treatment with rhG-CSF, when 70% of the donors could reach the peak CD34(+) cell yields. It is concluded that the age of the donors is the first factor determining the choice of donors for allogeneic hematopoietic stem cell transplantation, the sex, height, weight and BMI are secondary factors impacting yield of CD34(+) cells from donors mobilized with rhG-CSF.
Adolescent ; Adult ; Age Factors ; Antigens, CD34 ; immunology ; metabolism ; Blood Cell Count ; Blood Cells ; cytology ; immunology ; Body Height ; Body Mass Index ; Body Weight ; Child ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; Sex Factors ; Tissue Donors ; Young Adult

OBJECTIVETo compare the effects of pravastatin and granulocyto-colony stimulating factor (G-CSF) in mobilizing endothelial progenitor cells (EPCs) in mice with myocardial ischemia, and explore the possible mechanism of EPC mobilization.

METHODSNinety-six mice were randomly divided into 4 groups (n=24), namely the control group, saline group, pravastatin group and G-CSF group. In the latter 3 groups, myocardial ischemia was induced with isoprenine followed by intraperitoneal injections of normal saline, pravastatin and G-CSF for 5 consecutive days. On days 1, 5, 7, and 9 after establishment of myocardial ischemia, 6 mice from each group were randomly selected for measurement of the EPC count and serum concentrations of vascular endothelial growth factor (VEGF).

RESULTSCompared with the control group, EPC count increased slightly in the saline group on days 1, 5, and 7. EPC count increased significantly in pravastatin group on days 5, 7 and 9 in comparison with that of the saline group, and the increment was more obvious in G-CSF group. In comparison with the control group, the concentrations of VEGF augmented on days 5, 7 and 9 in the order of saline group, pravastatin group and G-CSF group. The effect of G-CSF on EPC mobilization was positively correlated to VEGF concentrations.

CONCLUSIONMyocardial ischemia induces EPC mobilization and VEGF release. Both Pravastatin and G-CSF can enhance EPC mobilization from the bone marrow and VEGF release, but G-CSF produces a stronger effect on EPC mobilization in association of VEGF release.

Animals ; Cell Movement ; drug effects ; Endothelial Cells ; drug effects ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Leukocyte Count ; Male ; Mice ; Myocardial Ischemia ; blood ; pathology ; Pravastatin ; pharmacology ; Stem Cells ; drug effects ; Time Factors ; Vascular Endothelial Growth Factor A ; blood