Adult ; Drug Users ; Female ; Humans ; Male ; Methadone ; administration & dosage ; therapeutic use ; Opioid-Related Disorders ; drug therapy ; epidemiology ; psychology ; Patient Acceptance of Health Care ; Quality of Life

creatinine concentration. Conclusion Bisphosphonates can decrease serum total calcium levels in hypercalcemia crisis caused by PHPT effectivelywith mild adverse events.

Objective to explore the current situation and related influencing factors on the retention time of patients receiving methadone maintenance treatment (MMT). Methods Information on basic situation and daily treatment of the patients were collected from the 7 MMT clinics opened in the pro-two batch in Hunan province. Retention rate and influencing factors were analyzed. Results (1) The retention rates after 6 and 12 months of MMT became 72.06% and 49.65% respectively. (2) The retention rates of high-dosage group and low-dosage group were 85.03% and 68.03% after 6 months on MMT program while became 60.48% and 46.28% after 12 months of MMT respectively. (3) The mean retention time of HIV+ patients and HIV- patients were 9.46 months and 8.62 months respectively during the 12 months follow-up observation, showing a significant difference. (4) Patients who took large dose methadone, did not share needles, at older age or HIV+ , were prone to keep MMT at a long period. Conclusion The retention rates for 6 months and 12 months in the MMT program in Hunan province were similar to the national data. Dose, type of drug abuse, age and HIV status were related to the period of retention.

OBJECTIVETo evaluate the expression of CD133 and its clinical significance in acute leukemia (AL) patients.

METHODSThe expression of CD133 and CD133 mRNA in leukemic blasts from 76 AL patients were detected by three-color flow-cytometry and hemi-quantitative RT-PCR respectively.

RESULTS(1) CD133 mRNA expression was highly correlated with CD133 expression in both of normal donors and AL patients groups. The expression of CD133 in AL patients was significantly higher than that in control group (P < 0.01). (2) The positive rates of CD133 and CD133 mRNA in AL group were 42.1% (32/76) and 46.1% (35/76) respectively. There was no significant difference in CD133 expression between AML-M(3) and normal control, AML and ALL, as well as T-ALL and B-ALL. The expression of CD133 in AML-M(4) were significantly higher than those in other AML subtypes (81.8% vs 43.7% and 81.8% vs 46.9% at CD133 and CD133 mRNA level, respectively, P < 0.01). (3) The expression of CD133 in AML was significantly correlated with the expression of CD34 and HLA-DR (P < 0.001). (4) The expression of CD133 had no relationship with the clinical prognostic factors such as cytogenetic or molecular aberrations, WBC counts, LDH, mdr1 expression and age. (5) There was a trend toward lower CR rate in CD133(+) cases, but only CD34/CD133(+) double positive cases had significant lower CR rate than that of negative ones (44.4% vs 71.4%, P < 0.05).

CONCLUSIONSAL had significantly higher CD133 expression compared to normal control. The detection of CD133 expression might help to identify AL type and predict therapeutic outcomes. Co-expression of CD133/CD34 might convey adverse prognosis of AL.

AC133 Antigen ; Acute Disease ; Adolescent ; Adult ; Aged ; Antigens, CD ; genetics ; Antigens, CD34 ; genetics ; Child ; Female ; Gene Expression Regulation, Neoplastic ; Glycoproteins ; genetics ; HLA-DR Antigens ; genetics ; Humans ; Leukemia ; genetics ; pathology ; Male ; Middle Aged ; Peptides ; genetics ; Prognosis ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo study the effect of ex vivo expansion on the migration ability and the CXCR4 expression of umbilical cord blood (CB) hematopoietic stem/progenitor cells (HSPC).

METHODSCD(34)(+) cells isolated from fresh CB samples were cultured in a serum-free and stroma-free culture system. On day 7, 10 and 14, CD(34)(+) cells were re-selected from the expanded cells, and the expression of CXCR4 and the transmigration ability of these CD(34)(+) cells were evaluated respectively and compared with those of the precultured fresh CD(34)(+) cells.

RESULTS(1) SDF-1 induced a higher migration percentage of fresh or expanded CB CD(34)(+) cells than that of uninduced ones. (2) Both of the uninduced and SDF-1-induced migrations were slightly reduced in the first week and then much more reduced in the second week expansion (P < 0.05). (3) The number of the CD(34)(+)CXCR4(+) cells were significantly increased during the culture period, but there was a downtrend of CXCR4 expression on CD(34)(+) subset; the expression levels on day 10 and 14 were lower than that on day 0.

CONCLUSIONSThe expanded HSPC would sustain the chemotactic activity during one-week-culture, but with further extended culture time their intrinsic homing potential would be partly impaired.

Antigens, CD34 ; genetics ; metabolism ; Cell Culture Techniques ; Cell Movement ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; metabolism ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Male ; Pregnancy

OBJECTIVETo explore the role of stromal cell-derived factor (SDF-1) and its specific receptor CXCR4 in the G-CSF-induced hematopoietic stem/progenitor cells (HSPCs) mobilization in human healthy donor.

METHODSThe changes of SDF-1/CXCR4 in bone marrow (BM) and peripheral blood (PB) of healthy donors during G-CSF-induced mobilization were detected by enzyme-linked immunosorbent assay (ELISA), immunohistological staining and flow cytometry. SDF-1 neutralizing antibody wes injected into BALB/c mice to further test its effect on mobilization.

RESULTSSDF-1 concentration in mobilized BM (mBM), steady state BM (ssBM) and PB were(7.23 +/- 0.66) microg/L, (5.43 +/- 0.35) microg/L and (5.42 +/- 0.52) microg/L, respectively. SDF-1 protein levels were decreased in the BM (P < 0.05) after 5-day G-CSF injection, and its concentration gradient between BM and PB disappeared (P > 0.05). Significant up-regulation of CXCR4 expression was observed on mBM CD34 cells in healthy donors. The rate of CXCR4 expression on CD34 cells in ssBM, mBM and mobilized PB were (40.98 +/- 21.56)%, (65.80 +/- 24.68)% and (27.54 +/- 26.03)%, respectively. Comparing with that in ssBM and mBM, CXCR4 expression on mobilized PB CD34+ cells were significantly decreased (P < 0.05). Inhibition of SDF-1 signal by blocking monoclonal antibodies significantly reduced G-CSF-induced mobilization in BALB/c mice. This resulted in significant decrease of white blood cell count and progenitors mobilized into peripheral circulation.

CONCLUSIONG-CSF induces HSPCs mobilization by decreasing bone marrow SDF-1 and down-regulating CXCR4 expression on HSPCs.

Animals ; Chemokine CXCL12 ; metabolism ; physiology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Receptors, CXCR4 ; metabolism ; physiology

OBJECTIVETo investigate the effect and the potential mechanism of umbilical cord (UC) derived mesenchymal stem cells (MSCs) on umbilical cord blood (UCB) derived CD34+ cells in vivo homing in xenotransplanted NOD/SCID mice model.

METHODSCD34+ cells and MSCs were derived from fresh UCB and UC, respectively. CD34+ cells (5 x 10(5) per mice) and MSC cells (5 x 10(6) per mice) were co-transplanted into irradiated NOD/SCID mice intravenously. CD34+ cells (5 x 10(5) per mice) alone were transplanted into the mice as control group. CD34+ cells home in bone marrow and spleen of recipient mice were detected 20 hours after transplant by FACS and RT-PCR, and the homing efficiencies were calculated. The effect of MSCs on CD34+ cells chemotactic function was investigated after co-cultured UCB CD34+ cells with UC MSCs in vitro. After 4 and 7 days coculture, the homing related adhesion molecules (the CD49e, CD31, CD62L, CD11a) expressed on CD34+ cells were detected by FACS.

RESULTS1) The homing efficiencies in bone marrow in experimental and control group were (7.2 +/- 1.1)% and (5.4 +/- 0.9)%, respectively (P < 0.05). 2) Human GAPDH gene was detected in bone marrow in experimental group and in spleen in both groups. 3) The migration efficiency of CD34+ cells was significantly higher in experimental group (35.7 +/- 5.8)% than in control group (3.5 +/- 0.6)% (P < 0.05). 4) The expression of CD49e, CD31, CD62L on CD34+ cells kept higher level in MSCs cocultured group than in CD34+ cells alone group.

CONCLUSIONSMSCs can efficiently increase homing of CD34+ cells to bone marrow and spleen in vivo by keeping a high level of homing adhesion molecules expression and improving migration efficiency of UCB CD34+ cells.

Animals ; Antigens, CD34 ; Cell Movement ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; metabolism ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Mice ; Mice, Inbred NOD ; Mice, SCID

OBJECTIVETo study the effect of ex vivo expansion on the adhesion activities of umbilical cord blood hematopoietic stem and progenitor cells (HSPC).

METHODSFresh UCB CD(34)(+) cells were cultured in a serum and stroma-free culture system. At day 7, day 10 and day 14, CD(34)(+) cells were re-selected from the expanded products. The expression of adhesion molecules (CAMs) such as VLA-4, VLA-5, LFA-1, ICAM-1, HCAM, L-selectin and PECAM-1, and the adhesion activity of the expanded CD(34)(+) cells were evaluated and compared with those of precultured fresh CD(34)(+) cells.

RESULTS(1) The CD(34)(+) cells expressing homing-related CAMs were increased (from 15-fold increase for CD(34)(+) CD(54)(+) subset to 72-fold increase for CD(34)(+) CD(49e)(+) subset at day 14). (2) The expressions of CD(49d), CD(44), CD(11a) and CD(49e) on the expanded CD(34)(+) cells were increased or sustained the same levels as those on fresh UCB CD(34)(+) cells, while the expression of CD(62L), CD(54) and CD(31) on expanded CD(34)(+) cells declined with the cultivating. (3) Spontaneous adhesion and SDF-1-induced adhesion tended to be increased in the course of the first 10 day's culture.

CONCLUSIONSThe culture system used in this study could substantially support the expansion of HSPCs expressing the above CAMs, and the expanded HSPCs would sustain their intrinsic adhesion potentials.

Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Cell Adhesion ; Cell Adhesion Molecules ; biosynthesis ; Cell Division ; Fetal Blood ; cytology ; immunology ; metabolism ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; metabolism ; Humans ; Receptors, Lymphocyte Homing ; biosynthesis

The purpose of this study was to explore the differences in megakaryocyte progenitor ex vivo expansion between CD34+ cells derived from human umbilical cord blood (CB) and bone marrow (BM). Mononuclear cells (MNCs) were obtained from CB or BM by Ficoll-Hypaque density gradient separation. CD34+ cells were purified by magnetic cell sorting (MACS). The selected CD34+ cells were seeded in serum-free conditions stimulated with thrombopoietin (TPO), TPO+interleukin 11 (IL-11), or TPO+IL11+heparin for 14 days. Amplification product (CD34+, CD41a+, and CD34+ CD41a+ cells) immunophenotypes, megakaryocyte apoptosis rates and the DNA content were measured by fluorescence-activated cell sorting (FACS). The colony-forming units of granulocytes and monocytes (CFU-GM), burst-forming units of erythrocytes (BFU-E), and colony-forming units of megakaryocytes (CFU-Mk) were also evaluated by the colony-forming units (CFU) assay. The results indicated that CD34+ cells derived from CB showed higher expansion ability of total cell counts, CD41a+ and CD34+ CD41a+ cells than those derived from BM for all days 14 of culture (p<0.05, respectively). There were no significant differences in CFU-GM, BFU-E, and total CFU-Mk counts between CB and BM-derived CD34+ cells on day 0 (p>0.05, respectively), but CB-derived CFU-Mk seemed mainly large colonies, and the number of large colonies was higher than that from BM (p<0.05) on day 0. There were no significant differences in expansion ability of CFU-GM between CB and BM-derived cells on days 7, 10, and 14 of culture (p > 0.05, respectively), but the expansion ability of BFU-E and CFU-Mk derived from CB cells was higher than that from BM (p<0.05, respectively). There were no significant differences in apoptosis rates of megakaryocyte from two source cells for days 14 of culture. Megakaryocytes derived from CB mostly showed the 2N DNA content (>90%) for days 14 of culture, while those cells derived from BM showed the increased DNA content, and 4N, 8N or more ploidy cells gradually increased with prolonging of culture time. It is concluded that CB-derived CD34+ cells have a greater proliferation potential than that derived from BM, which is therefore proven to be a better cell source for megakaryocyte progenitor expansion in vitro.
Antigens, CD34 ; Bone Marrow Cells ; cytology ; immunology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Division ; Cell Separation ; Cells, Cultured ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; immunology ; Humans ; Megakaryocyte Progenitor Cells ; cytology ; immunology

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism