The aim of study was to investigate the modulation effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on adhesion molecule expression of memory T lymphocyte in the bone marrow grafts. rhG-CSF was administered in 41 donors by subcutaneous injection for 5 consecutive days. Bone marrow grafts were collected on day 4. The percentages of CD4+ and CD8+, and the expressions of CD49d, CD54, CD62L and CD11a on donor T cells of steady state-bone marrow grafts (SS-BM, n=11) and rhG-CSF primed bone marrow (G-BM, n=30) were analyzed by using multi-color flow cytometry. The results indicated that the percentages of CD4+ and CD8+ T cells were significantly lower in G-BM than those in SS-BM (p<0.05). There were no significant differences in the percentages of memory CD4+ and CD8+ T cells between SS-BM and G-BM (p>0.05). The expressions of CD49d on CD4+ and CD8+T cells were significantly lower in G-BM than that in SS-BM (p<0.05). Compared with SS-BM, the expressions of CD54 on CD4+, memory CD4+ T cells and CD8+ T cells were significantly lower in G-BM (p<0.05). The expressions of CD62L on CD4+ and CD8+ T cells and memory T cells were all significantly lower in G-BM (p values were all less than 0.001). The expressions of CD11a on CD4+, memory CD4+ T cells were significantly lower in G-BM than that in SS-BM (p<0.05). There were no significant differences in the expression of CD11a on CD8+, memory CD8+ T cells between SS-BM and G-BM (p>0.05). It is concluded that the expression of cell adhesion molecules on the CD4+ and CD8+ T cells in G-BM is down-regulated after rhG-CSF treatment of healthy donors.
Adolescent ; Adult ; Aged ; Bone Marrow ; Bone Marrow Cells ; immunology ; metabolism ; Bone Marrow Transplantation ; immunology ; methods ; Cell Adhesion Molecules ; metabolism ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Humans ; Living Donors ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocytes ; immunology ; metabolism ; Young Adult

The study was purposed to investigate the effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in healthy donors after recombinant human granulocyte colony-stimulating factor (rhG-CSF) application in vivo. Sixty-two healthy donors were treated with rhG-CSF [5 microg/(kg.d)] injected subcutaneously for five consecutive days. Donors were divided into groups A and B, and 31 donors were there in each group. Bone marrow grafts and peripheral blood stem cell grafts were harvested on the day 4 and 5 respectively in group A. In group B, only peripheral blood grafts were collected on both the day 4 and 5. The quantities of the cell components, CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry. The results showed that median counts of nuclear cells (1.56 x 10(5)), lymphocytes (8.56 x 10(4)), CD3(+) (6.12 x 10(4)), CD3(+)CD4(+) (3.38 x 10(4)), CD3(+)CD8(+) (2.27 x 10(4)), CD14(+) (3.83 x 10(4)), CD34(+) cells (744) and CD3(+)CD4(-)CD8(-) T cells (3588) per microliter of peripheral blood stem cell grafts in group A were similar to counts of nuclear cells (1.40 x 10(4)), lymphocytes (7.34 x 10(4)), CD3(+) (5.32 x 10(4)), CD3(+)CD4(+) (3.06 x 10(+)), CD3(+)CD8(+) (1.83 x 10(4)), CD14(+) (3.21 x 10(4)), CD34(+) cells (554) and CD3(+)CD4(-)CD8(-) T cells (3120) in group B (p > 0.05). There were no difference in the ratios of CD4(+) cells/CD8(+) cells, CD14(+) cells/CD3(+) cells and CD3(+)CD4(-)CD8(-) T cells/CD3(+) cells in peripheral blood stem cell grafts between group A [1.52 (0.54 - 2.87)], [0.57 (0.15 - 1.64)], [0.064 (0.018 - 0.673)] and group B [1.68 (0.31 - 3.35)], [0.59 (0.18 - 1.25)], [0.063 (0.021 - 0.136)] (p > 0.05). It is concluded that no effect of bone marrow graft collection on the composition of peripheral blood stem cell grafts in the same donor is found after rhG-CSF application in vivo, bone marrow grafts and peripheral blood stem cell grafts can be collected respectively or simultaneously.
Adolescent ; Adult ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Tissue Donors ; Young Adult

The study was aimed to analyze the difference of naive and memory CD4(+) and CD8(+) T-cell subsets between steady-state bone marrow (SS-BM) and recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood grafts (G-PB). Four CD4(+) and CD8(+) T-cell subsets classified according to the expression of CD45RA and CD62L were determined by three-color flow cytometry. The results showed that the percentage of CD4(+), CD8(+) T-cell subsets and the ratios of CD4/CD8 in G-PB were significantly higher than those in SS-BM (p < 0.05). The percentage of CD4(+) naive T-cells in G-PB was significantly lower than that in SS-BM (p < 0.001). As compared with SS-BM, the percentage of CD4(+) effector memory T-cells was significantly high in G-PB (p < 0.001). There were no significant differences in the percentages of the four CD8(+) T-cell subsets between SS-BM and G-PB (p > 0.05). The percentage of CD4(+)CD62L(+) T-cells in G-PB was significantly lower than that in SS-BM (p = 0.001). The absolute numbers of CD4(+) and CD8(+) T-cell subsets, the eight naive and memory CD4(+) and CD8(+) T-cell subsets were significantly higher in G-PB than those in SS-BM (p < 0.001). It is concluded that the difference of naive and memory CD4(+) and CD8(+) subsets between G-PB and SS-BM may partially explain why the incidence and severity of acute graft-versus-host disease (GVHD) was similar and the incidence of chronic GVHD was different after transplantation with SS-BM or G-PB.
Adolescent ; Adult ; Aged ; Bone Marrow Transplantation ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Young Adult

OBJECTIVETo investigate the regular pattern of Cytomegalovirus (CMV)-specific T cells (CTL) immune reconstitution after human leukocyte antigen (HLA) matched sibling donor allogeneic bone marrow(BM) plus peripheral blood hematopoietic stem cell (PBSC) transplantation.

METHODSCTL from seventeen patients after transplantation was detected by flow cytometry, the IFN-γ secretion ability of CTL by enzyme-linked immunospot (ELISPOT) assay, and clonal analysis of TCR Vβ subfamily by gene scan assays. The relationship between CTL reconstitution and CMV infection was studied.

RESULTSBoth number and function of recipients CTL reached to normal control level at 30 d post-transplantation. The recipients achieved a high frequency CTL with IFN-γ response and restoration of T-cell receptor β (TCR Vβ) repertoire at one year post-transplantation. CTL with the central memory CD45RO(+)CD62L(+) cell phenotype expanded in PB when CMV was reactivated. The incidence of CMV reactivation was 35.83% (17.91% - 63.10%) after transplantation, and none of them developed CMV disease.

CONCLUSIONAfter HLA matched related donor transplantation using mixed grafts, immune recovery to CMV seems to be early and fast. The incidence of CMV infection and disease are low.

Adult ; Cytomegalovirus ; immunology ; Female ; HLA Antigens ; immunology ; Hematologic Diseases ; immunology ; surgery ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Siblings ; T-Lymphocytes, Cytotoxic ; immunology ; Tissue Donors ; Young Adult

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 purposed to investigate the relation of monocyte counts in peripheral blood (PB) at the first collection of allograft to the amount of CD34(+) cells in the mixture of recombinant human granulocyte colony-stimulating factor (rhG-CSF)-primed bone marrow graft (G-BM) and rhG-CSF mobilized peripheral stem cell grafts (G-PB). 70 healthy donors were treated with rhG-CSF [5 microg/(kg.d)] injected subcutaneously for 5 consecutive days. Bone marrow grafts and peripheral blood grafts were harvested on the 4th and 5th days respectively. Blood cell counts at the first collection of allografts were determined by blood analyzer XL2100, the amount of CD34(+) cells in G-BM and G-PB were determined by flow cytometry. The results showed that the monocyte counts in the PB of the 70 healthy donors were (1.15 +/- 0.60) x 10(9)/L. The amount of total CD34(+) cells in the G-BM, G-PB and mixture grafts were (5.85 +/- 2.93) x 10(7), (1.33 +/- 0.77) x 10(8), and (1.92 +/- 0.86) x 10(8) respectively. Pearson and Spearman correlation analysis indicated that the counts of monocyte at the first collection of allografts correlated positively with the amount of total CD34(+) cells in the G-BM (correlation coefficient, r = 0.265, p = 0.027), G-PB (r = 0.340, p = 0.004) and mixture grafts (r = 0.398, p = 0.001). Stepwise Linear Regression Model analysis also showed that the counts of monocytes in the PB correlated positively with the amount of CD34(+) cells in the G-BM, G-PB and mixture grafts (p = 0.027, 0.004 and 0.001 respectively). The sensitivity and specificity of the monocyte counts to predict the amount of CD34(+) cells in the mixture grafts were 71% and 70% respectively (p = 0.007). In conclusion, the monocyte counts in the PB at the first collection of allografts after rhG-CSF treatment of healthy donors may be a simple and practical indicator for harvest of CD34(+) cell amount in the mixture grafts.
Adolescent ; Adult ; Antigens, CD34 ; immunology ; Blood Cell Count ; Bone Marrow Transplantation ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Monocytes ; cytology ; immunology ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Tissue Donors ; Young Adult

OBJECTIVETo investigate the cell composition in granulocyte colony-stimulating factor One (rhG-CSF) primed bone marrow grafts (G-BM) from donors with different characteristics.

METHODShundred and fifty healthy donors were injected subcutaneously rhG-CSF (5 microg x kg(-1) x d(-1)) in five consecutive days. Bone marrow and peripheral blood stem cells were harvested at day 4 and day 5. The number of CD3, CD3+ CD4+, CD3+ CD8+, CD14+ , CD34+ and CD3+ CD4- CD8- cells were determined by multicolor flow cytometry. Cell composition of G-BM from donors with different characteristics, including sex, age, weight, pregnancy were analysed.

RESULTSThe absolute number of nuclear cells (NCs), lymphocytes, CD3+, CD3+ CD4+, CD3+ CD8+, CD14+, CD34+ and CD3+ CD4- CD8- cells in G-BM were 31 050 (12 200 -58 100), 2122 (506 - 6618), 1344 (307 - 4791), 692 (145 - 3038), 616 (112 - 2575), 986 (265 -2958), 63 (11 -505) and 83 (9 -390) per microliter, respectively. The number of NCs [33 800 (18 600 - 57 100)], CD34+ cells [76 (22 -505)], and CD3+ CD4+ CD8- regulatory T cells [97 (11 - 380)] harvested from younger donors (aged < or = 38 years) were significantly higher than those from older ones (aged > 38) [28000 (12200-58100), 49 (11-220), and 65 (9 - 389) per microliter (P = 0.027, < 0.001 and = 0.001, respectively)]. Compared with that in donors with peripheral blood monocytes (PBMs) < or = 0.37 x 10(9)/L, higher number of NCs in G-BM [27 150 (13 800 - 58 100) vs 33 550 (12 200 - 57 100), P = 0.005] were collected in donors with PBMs > 0.37 x 10(9)/L. Multivariate analysis showed that donors age (< or = 38 vs > 38 years; P = 0.01) and monocyte number in peripheral blood (< or = 0.37 x 10(9)/L vs > 0.37 x 10(9)/L; P = 0. 003) were factors predicting for NC yields, and donors age ( < or = 38 vs > 38 years; P < 0.001) was factor predicting for yields of CD34+ cells (P < 0.001) and CD3+ CD4- CD8- regulatory T cells (P = 0.001) collection.

CONCLUSIONDonor age is a factor for the yields of NCs, CD34+ cells, and CD3+ CD4- CD8- regulatory T cells collection in G-BM, and donor's PBMs more than 0.37 x 10(9)/L, is another factor affecting NCs harvests.

Adolescent ; Adult ; Age Factors ; Antigens, CD34 ; Bone Marrow Cells ; immunology ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; Tissue Donors ; Young Adult

OBJECTIVETo investigate the feasibility, effectiveness and practicability of transurethral enucleation plus pneumocystostomy rotary cut (TUE + PCRC) for large benign prostatic hyperplasia (BPH).

METHODSWe performed TUE + PCRC for 26 BPH patients aged 62 - 85 years with the prostate volume of 80 - 165 ml. We conducted transurethral enucleation of the hyperplastic prostate glands and pushed them into the bladder, followed by bladder puncture for pneumo-cystostomy rotary cut.

RESULTSAll the surgical procedures were successfully accomplished, with the mean surgical time of 41 (32 - 54) minutes and intraoperative blood loss < 60 ml in all the cases. Twenty-three of the patients were followed up for 2 - 8 months, which revealed no stricture of the urethra or any other severe complications. Compared with the preoperative baseline, significant improvement was achieved in the IPSS (6.5 +/- 2.2 vs 26.2 +/- 2.4), QOL (1.4 +/- 0.9 vs 4.6 +/- 1.2) and Qmax ([5.8 +/- 1.0 ] vs [19.6 +/- 2.8] ml/s) of the patients after surgery (P < 0.01).

CONCLUSIONTUE + PCRC, with its advantages of short operation time and less severe complications, is a safe and effective approach to the management of large BPH.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; surgery ; Transurethral Resection of Prostate ; methods

OBJECTIVETo investigate the association of the HLA-DRB1 polymorphism with susceptibility to myelodysplastic syndrome (MDS) and aplastic anemia (AA) in Chinese Han population.

METHODSThe polymorphism of HLA-DRB1 alleles in 242 patients with MDS, 115 patients with AA and 2264 umbilical cord blood control samples were tested by polymerase chain reaction-sequence specific primer (PCR-SSP).

RESULTSCompared with normal controls, the frequency of HLA-DRB1*15 was significantly increased in the MDS group and AA group (22.93% vs. 17.25%, Chi-square = 9.662, OR= 1.428, P= 0.003; 26.52% vs. 17.25%, Chi-square = 12.924, OR= 1.732, P= 0.001). And this is mainly due to the increase in the male patients in both patient groups: in the MDS males, 24.68% vs. 17.25%, Chi-square= 11.194, OR= 1.572, P= 0.001, and in the AA males, 29.29% vs. 17.25%, Chi-square= 13.563, OR= 1.987, P= 0.001. No significant difference between the controls and the female patients in both groups was observed.

CONCLUSIONThe HLA-DRB1*15 could be a susceptibility allele for MDS and AA male patients.

Alleles ; Anemia, Aplastic ; genetics ; Asian Continental Ancestry Group ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Myelodysplastic Syndromes ; genetics ; Polymorphism, Genetic

The objective of this study is to develop an in vitro screening method for nasal absorption of insulin. First, the adaptability of in situ rat nasal perfusion test for the study of insulin was investigated. It was found that insulin was liable to be absorbed on the silicone tube and the traditional method is not suitable. However, addition of 0.001% Labrasol into the perfusate can effectively solve this problem. A modified method suitable for in situ rat nasal perfusion of insulin was established with the addition of 0.001% Labrasol into the perfusate. Using the modified method, effect of pH and drug concentration on the absorption of insulin in the nasal cavity was further investigated. The results suggest that compared with pH 4.5 and pH 7.4, the drug absorption rate was the lowest at pH 6.0. The intranasal absorption mechanism of insulin may be passive diffusion.