Apoptosis ; Female ; Gardening ; Humans ; Leukocytes ; drug effects ; pathology ; Male ; Occupational Exposure ; adverse effects ; Pesticides ; adverse effects

The anticancer agent's FT-207, N1-(2'-tetrahydrofuryl)-5-fluorouracil, a derivative of 5FU (5-fluorouracil), induced chromosome damage to the human leukocyte was investigated. FT-207 inhibit mitosis and cause chromatid and chromosome breakage and chromatid exchange with 20 ug/ml for 48 to 72 hours of treatment. However, with 15 ug/ml for 72 hours only delayed spiralization was produced in some of the chromosomes in the same cells. The random distribution of chromosome breakage were observed and the effect of FT-207 on the chromosomes of human leukocytes were time dependent rather than concentration dependent. The comparision of the effect of mitomycin C on human leukocytes and the action of FT-207 at specific times during the cell cycle were discussed.
Cells, Cultured ; Chromosome Aberrations* ; Female ; Fluorouracil/analogs & derivatives* ; Human ; Leukocytes/drug effects ; Leukocytes/ultrastructure* ; Male ; Mitotic Index/drug effects ; Tegafur/pharmacology* ; Time Factors

OBJECTIVETo investigate effective doses of cyclophosphamide (CTX) for establishment of leukopenia model in ICR mouse.

METHODSNormal ICR mice (n = 96) were divided into CTX(80), CTX(100), CTX(120), CTX(150) and CTX(300) groups, of which mice received CTX intraperitoneally at a dose of 80, 100, 120, or 150 mg/kg once a day for 3 consecutive days, or 300 mg/kg with single injection. The peripheral blood cell counts were detected at various times before and after CTX administration.

RESULTSThe peripheral white blood cell nadirs in the mice injected with CTX appeared on day 4 after the first dose of CTX. The peripheral white blood cell nadir in group CTX(100) was 26.7% of the value measured in mice before CTX administrated, and that of group CTX(80) was 35.0%. Higher doses of CTX, however, caused too severity in hematopoietic injury.

CONCLUSIONThe dose of CTX 100 mg/(kg·d) × 3d is appropriate for leucopenia model of ICR mouse.

Animals ; Blood Cell Count ; Cyclophosphamide ; administration & dosage ; Leukocytes ; drug effects ; Mice ; Mice, Inbred ICR

Animals ; Leukocyte Count ; Leukocytes ; drug effects ; Male ; Mice ; Nerve Growth Factor ; pharmacology ; Skin ; pathology ; radiation effects ; Survival Rate ; Whole-Body Irradiation ; adverse effects ; mortality ; Wound Healing ; drug effects

OBJECTIVETo investigate the influence of high-voltage electrical burn (HEB) on the aggregation and adhesion of platelet and leukocyte in rats and the interventional effect of pentoxifylline (PTX).

METHODSOne hundred and eighty SD rats were divided into control, electrical burn (EB), and pentoxifylline treatment (PT) groups according to the random number table, with 60 rats in each group. (1) Ten rats were taken from each group at 15 minutes before injury for the observation of the microcirculatory perfusion of chest skin with Laser Doppler Perfusion Imager (LDPI), and the number of leukocyte adherent to mesenteric venule with Bradford Variable Projection Microscope (BVPM). Serum was collected from heart blood to determine the contents of platelet activating factor (PAF), thromboxane B2 (TXB2), prostacyclin (PGI2), P-selectin, E-selectin and L-selectin by double-antibody sandwich enzyme-linked immunosorbent assay. The ratio of TXB2 to PGI2 was calculated therefrom. (2) Model of HEB was reproduced in the remaining 50 rats of EB group and that of PT group with voltage regulator and experimental transformer (the electrical current applied to the left forelimb and exited from the right hind limb). The remaining 50 rats of control group were sham injured with the same devices without electric current. Within 2 minutes post injury (PIM), rats in control group and EB group were intraperitoneally injected with 2 mL isotonic saline, while rats in PT group were intraperitoneally injected with 2 mL pentoxifylline (50 mg/mL). At PIM 5 and 1, 2, 4, 8 hour(s) post injury (PIH), 10 rats of every group were randomly chosen at each time point for the observation of the microcirculatory perfusion of chest skin and the number of leukocytes adherent to mesenteric venule through the same method as used above, and the levels of the related factors of aggregation and adhesion of platelets and leukocytes were determined, and then the relative ratio was calculated. Data were processed with the analysis of variance of factorial design and LSD test.

RESULTSThe contents of PAF, TXB2, PGI2, P-selectin, E-selectin, L-selectin, and the ratio of TXB2 to PGI2, as well as the number of adhered leukocyte in EB group were higher, while the microcirculatory perfusion value was lower than those of control group, with F values from 854.20 to 8156.52, P values all below 0.01. The microcirculatory perfusion value and PGI2 content of PT group were higher, while the contents or number of other indexes were lower than those of EB group, with F values from 33.18 to 1033.99, P values all below 0.01. Only the data within EB group and PT group were comparable. The contents of PAF, TXB2, PGI2, P-selectin, E-selectin, L-selectin, and the ratio of TXB2 to PGI2, as well as the number of adhered leukocyte in EB group and PT group at each time point were significantly higher than those at 15 minutes before injury, while the microcirculation perfusion value was significantly lower than that at 15 minutes before injury (P values all below 0.001), with the exception of the ratio of TXB2 to PGI2 in PT group and E-selectin in EB group and PT group at PIM 5. The contents of PAF, TXB2, and E-selectin and the ratio of TXB2 to PGI2 in EB group peaked at PIH 4, and they were respectively (9.3 ± 0.9) ng/mL, (14.31 ± 0.65) nmol/mL, (271.2 ± 18.4) ng/mL and 4.62 ± 0.26. The contents of PGI2 and P-selectin, and the number of adhered leukocyte in EB group peaked at PIH 8, and they were respectively (3.98 ± 0.24) nmol/mL, (514 ± 24) ng/mL, and (25.50 ± 4.14) per 100 µm venule. The content of L-selectin peaked at PIH 2 [(876 ± 54) ng/mL]. The microcirculatory perfusion value was lowest at PIM 5 [(1.17 ± 0.10) V].

CONCLUSIONSHEB can increase the contents of PAF, TXB2, PGI2, P-selectin, E-selectin, L-selectin, the ratio of TXB2 to PGI2, and the number of adhered leukocyte, as well as decrease the skin microcirculatory perfusion value. PTX can inhibit the aggregation and adhesion of platelets and leukocytes through increasing the content of PGI2 and decreasing contents of other factors mentioned above, thus alleviating the microcirculatory dysfunction after HEB.

Animals ; Blood Platelets ; drug effects ; Burns, Electric ; blood ; physiopathology ; Leukocytes ; drug effects ; physiology ; Male ; Pentoxifylline ; pharmacology ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley

The effects of erythropoietin and recombinant cytokines (G-CSF, SCF, IL-3 and GM-CSF) on colony formation and self-renewal by erythroid burst-forming units (BFU-E) and granulocyte-macrophage progenitors (CFU-GM) from mobilized peripheral blood progenitor cells (PBPCs) of the normal donors and patients were investigated. To better understand how combinations of the cytokines may be appropriate for stem cell manipulation, a model involving replating of individual BFU-E and CFU-GM colonies has been used to study the effects of cytokines on colony formation and self-renewal. Based on granulocyte colony-stimulating factor (G-CSF) and erythropoietin(EPO) alone serve as a baseline with which to compare the effects of the combinations with stem cell factor (SCF), interleukin-3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF). The results revealed that: 1. BFU-E derived from PBPCs produced a significantly great numbers of subcolonies in EPO plus IL-3 and EPO+SCF+IL-3, and a difference in EPO+SCF compared with EPO alone. 2. Compared patients to normal donors the self-renewal ability of BFU-E was not influenced after EPO plus SCF or IL-3. 3. There was a significantly increased frequency of CFU-GM in the presence of SCF, IL-3, and GM-CSF used in conjunction with G-CSF. Comparing the frequencies of CFU-GM in patients and donors, patients' PBPCs were more sensitive to G-CSF+SCF and GMix (G-CSF+SCF+IL-3+GM-CSF), especially to G-CSF alone than donors. 4. There was a significant increase in AUC (area under the curve of subcolony distribution) in the presence of G-CSF combined with other cytokines. GMix was identified as the optimal combination of cytokines for both the expansion of CFU-GM as well as the expansion of clonogenic progenitor cells. 5. Donors have a high AUC than patients, especially there was a significant increase AUC in G-CSF alone (P = 0.0067).
Cytokines ; pharmacology ; Erythroid Precursor Cells ; drug effects ; Erythropoietin ; pharmacology ; Humans ; Leukocytes, Mononuclear ; drug effects ; physiology ; Recombinant Proteins ; pharmacology ; Stem Cells ; drug effects

Adult ; Apoptosis ; drug effects ; Benzene ; poisoning ; Biomarkers ; analysis ; Bone Marrow Cells ; drug effects ; Female ; Flow Cytometry ; Humans ; Leukocytes, Mononuclear ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Micronuclei, Chromosome-Defective ; drug effects ; metabolism ; Occupational Exposure ; analysis

This study was aimed to investigate the effect of dexamethasone (Dex) on immunosuppressive ability of mesenchymal stem cells (MSC) during expansion and differentiation of MSC. MSC were cultured in 96-well flat-bottom plates. Proliferation assays were performed by using the BrdU colorimetric ELISA Kit. To explore the effect of Dex on MSC immunosuppressive ability, MSC were firstly cultured in complete culture medium for 14 d with Dex (10 nmol/L), and then, peripheral blood mononuclear cells (PBMNC) were co-cultured with MSC in 96-well flat-bottom plates for 3 d. Phytohemagglutinin A (PHA, 10 µg/ml) was used to stimulate activation of PBMNC. The concentrations of IFN-γ in culture supernatants was detected by ELISA. The results indicated that there was no obvious difference in representative phenotypes of MSC between experimental and control groups after MSC were treated with low concentration of Dex (10 nmol/L) for 14 d, but the suppression of Dex-treated MSC on lymphocyte activation in same concentration of cells was significantly reduced as compared with control group. After the Dex-treated MSC were co-cultured with IFN-γ for 12 h, the immunoregulatory ability of MSC was recovered in a certain degree. It is concluded that the Dex impairs the immunosuppressive ability of MSC, the IFN-γ can protect and reverse the immunosuppressive ability of MSC impaired by Dex, so that, when the immunoregulatory activity of MSC is investigated, it is necessary to avoid adding Dex in the culture medium.
Cells, Cultured ; Dexamethasone ; adverse effects ; Humans ; Immune Tolerance ; drug effects ; Interferon-gamma ; immunology ; Leukocytes, Mononuclear ; Lymphocyte Activation ; immunology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; immunology

OBJECTIVETo study the effect of hydroquinone on apoptosis of bone marrow mononuclear cells, and to evaluate the toxic effect of benzene on stem cells.

METHODSCell morphology was observed by HT fluorescent stain method, and DNA fragments were analyzed by agarose gel electrophoresis. Anti-Annexin V FITC plus PI staining for apoptotic and necrotic rate was examined by flow cytometer.

RESULTSAfter adding different concentrations of hydroquinone to the cells for 6 h culture, the fluorescent intensity of nucleus increased, the color of nucleus became deep and inhomogeneous, and the chromatin was condensed and distributed around the neucleus. DNA ladder was detected in all samples. Cell apoptotic rate in different concentration of hydroquinone groups was significantly higher than that in blank control group (P < 0.05). With the increase of the concentration of hydroquinone, the apoptotic and necrotic rate also increased. The optimal concentration of hydroquinone was 50 micro mol/L. When it was >or= 75 micro mol/L, the necrotic rate increased significantly. Hydroquinone-induced apoptosis was associated with culture time at the concentration of 50 micro mol/L, and the peak apoptotic time was 10 h, then the apoptotic rate decreased and necrotic rate increased.

CONCLUSIONHydroquinone can induce apoptosis of bone marrow mononuclear cells in vitro with dose-effect and time-effect relationship.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Hydroquinones ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; Mutagens ; pharmacology

OBJECTIVETo investigate the feasibility of prevention and treatment of early scar through observing the effect of feeding immunosuppressive drug cyclophosphamide on rabbit ears hypertrophic scar tissue in early stage.

METHODSThirty-two Rabbit ears were used to establish animal models for hypertrophic scar and randomly divided into four groups: group of distilled water (A), group of cyclophosphamide 5 mg x kg(-1) x d(-1) (B), group of 10 mg x kg(-1) x d(-1) (C), group of 30 mg x kg(-1) x d(-1) (D). Before animal models were built and after administration for 14 days, 28 days, leukocytes and lymphocytes were detected. After 28 days, specimens were harvested and underwent HE staining and VG staining in order to assess HI, NA, AA value changes. The data (HI, NA, AA) from each group were compared by analysis of variance, and the variance for the rank sum test when missing.

RESULTSOn the 14th day, the number of leukocytes in group A, B, C, D were (8.62 +/- 0.58) x 10(9)/L, (4.48 +/- 0.41) x 10(9)/L, (2.7 +/- 0.26) x 10(9)/L, (1.33 +/- 0.27) x 10(9)/L; the number of lymphocytes in group A, B, C, D were (3.11 +/- 0.21) x 10(9)/L, (1.67 +/- 0.16) x 10(9)/L, (0.42 +/- 0.10) x 10(9)/L, (0.40 +/- 0.09) x 10(9)/L. On the 28th day, the number of leukocytes in group A, B, C, D was (8.63 +/- 0.53) x 10(9)/L, (5.10 +/- 0.27) x 10(9)/L, (3.10 +/- 0.26) x 10(9)/L, (1.98 +/- 0.20) x 10(9)/L; the number of lymphocytes A, B, C, D was (3.06 +/- 0.16) x 10(9)/L, (2.08 +/- 0.14) x 10(9)/L, (0.96 +/- 0.19) x 10(9)/L, (0.14 +/- 0.07) x 10(9)/L. On the 14th day and 28th day, the number of leukocytes and lymphocytes in experimental groups was reduced, showing a negative relation with cyclophosphamide dose (P < 0.05). The HI in group of A, B, C, D was 3.02 +/- 0.24, 2.59 +/- 0.43, 2.06 +/- 0.19, 1.63 +/- 0.11; the AA was 40.49 +/- 2. 07, 35.29 +/- 1.99, 28.36 +/- 1.87, 24.99 +/- 1.82; the NA was 4570.5 +/- 259.3, 4222.5 +/- 199.6, 3540.3 +/- 170.3, 3341.4 +/- 228.8. The difference in HI, AA, NA between control group and any of the experimental groups was statistically significant (P < 0.01). Each group, with the dose increased, except NA content of group C and D, the HI, AA, NA was more smaller, negative correlation, the difference was statistically significant (P < 0.05).

CONCLUSIONSFeeding cyclophosphamide can inhibit leukocytes and lymphocytes number, so as to inhibit the proliferative activity of hypertrophic scar. It has significant effect on prevention of hypertrophic scar on rabbit ears in early stage.

Animals ; Cicatrix, Hypertrophic ; drug therapy ; prevention & control ; Cyclophosphamide ; pharmacology ; Ear ; pathology ; Female ; Leukocyte Count ; Leukocytes ; drug effects ; Lymphocyte Count ; Lymphocytes ; drug effects ; Male ; Rabbits