AIM: To characterize the gene expression of sortilin on adipogenic and osteogenic differentiation in mesenchymal stem cells (MSCs) in vitro and explore its significance.METHODS: MSCs derived from human bone marrow were isolated and cultured in vitro ,then were stimulated in osteogenic medium and adipogenic medium,respectively. Osteopontin and lipoprotein lipase were detected by RT-PCR. Sortilin expression was analyzed by semiquantitative RT-PCR. RESULTS: 1.MSCs displayed the potential of differentiation into osteoblast and adipocyte. 2.Sortilin was upregulated one day after osteogenic induction and remained upregulated for a week. The expression of sortilin was significant increased on day 3( P 0.05).CONCLUSION: Sortilin may be useful to modulate the osteogenic differentiation and may not be necessary for adipocyte commitment in MSCs. The regulation of sortilin expression may provide new protocal and strategy for the treatment of osteoporosis and osteopenic disease.

AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-?B activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P

AIM: To investigate the effects of transforming growth factor ?1 (TGF-?1) on murine-derived dendritic cells (DC). METHODS: Murine bone marrow cells were cultured with GM-CSF and TGF-?1 to develop TGF ?-DC. Then they were stimulated by lipopolysaccharide (LPS). Their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using BrdU ELISA method. IL-12 p70 protein was detected by ELISA and the expressions of Toll like receptor 4 (TLR4) on DCs were measured by semi-quantitative RT-PCR and FCM. RESULTS: Compared to immature DC (imDC) cultured with GM-CSF alone, the expressions of CD_80, CD_86, I-A~b and CD_40 in TGF ?-DC were lower. The TGF ?-DC was resistant to maturation by LPS. Maturation resistance was evident from a failure to up-regulate CMs, to stimulate larger T cell proliferation and to increase secretion of IL-12 p70. Down-regulation of TLR4 expression on TGF ?-DC was also found. CONCLUSION: TGF-?1 inhibits the expression of co-stimulatory molecules on DC. It is resistant to maturation stimulus (LPS) and might be linked with TLR4 down-regulation.

Objective To study the influence of sunscreens with different efficacy on delayed type hypersensitivity (DTH) and their immunoprotective effect in mice.Methods A cohort of mice were randomly divided into 5 groups with 10 mice in each group:group 1 as the positive control without irradiation,group 2 receiving solar-simulated radiation (SSR) only,group 3 receiving SSR and protected by sunscreen l with sun protection factor 15(SPF15)and persistent pigment darkening(PPD)12,group 4 receiving SSR and protected by sunscreen 2 with SPF 50 and PPD 28,and group 5 as the negative contml receiving SSR only.SSR was carried out on the back of mice with the UVA dose being 1.4 J/cm2 and UVB dose being 100 mJ/cm2 for 10 days.After a 5-day irradiation,the groups 1 to 4 were immunized by intraperitoneal injection with 100 μl(107 cells/ml) of Candida albicans suspension.On the 10th day both sides of the posterior foot pad were measured;then the foot pads were injected with additional 50 μl of the Candida albicans suspension.Twenty-four hours after the injection,the thickness of each foot pad was measured,and immunosuppression rate was calculated.Finally,the mice were sacrificed and skin samples were obtained from the back of these mice followed by the examination of CDla, CD80 and CD86 expression by Western blot.Resets The thickness of edema in foot pads was 0.41±0.38 mm,0.21±0.23 mm and 0.30 ± 0.25 mm in group 1,3 and 4,respectively,significantly higher than in group 5 and 2(0.04±0.03 mm,0.14±0.12 mm,respectively,all P<0.05),while no significant difference was observed between the group 3 and 4(P>0.05).Significant differences were observed in the immunosuppression rate between group 2,3 and 4(73.0%±11.3%,54.1%±6.4%,29.7%±7.5%,respectively,all P<0.01).Western blot revealed a significant increment in the expression of CDla protein in group 1 compared with group 2 as well as in the expression of CD86 protein in group 1 and group 3 compamd with group 2 and group 5(all P<0.05),but no statistical difference was observed between the other groups in the expression level of CDla,CD80 or CD86(P>0.05).Conclusions The exposure to sub-erythema dose of UV can induce DTH,and sunscreens have an immunoprotective effect in this process.Epidermal Langerhans cells are not essential for UV-induced immunosuppression.

OBJECTIVETo evaluate the effect of mycophenolate mofetile (MMF) on acute graft versus host disease (aGVHD) prophylaxis after allogeneic bone marrow transplantation (allo-BMT) in a murine model.

METHODSAcute GVHD was induced in male BALB/c mice by total-body irradiation (TBI) followed by female allogeneic (C57BL/6J) bone marrow and spleen cells transplantation. Acute GVHD was assessed both physically and histologically. Severe aGVHD was developed in the recipients and the mean survival time (MST) of untreated BM recipients was 6 days. After allo-BMT, animals were divided into 6 groups. Group 1 was given MMF (30 mg.kg(-1).d(-1)), group 2 CsA (1.5 mg.kg(-1).d(-1)) and MTX (0.4 mg.kg(-1).d(-1)), group 3 MMF (30 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 4 MMF (60 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 5 MMF (10 mg.kg(-1).d(-1)) in combination with CsA and MTX, and group 6 no agents for aGVHD prophylaxis. The physical signs, MST, peripheral blood counts, and aGVHD histopathologic examination were observed in all recipients.

RESULTSAnimals in control group developed typical aGVHD and 100% of mortality, with a MST of 6 days. The MSTs of group 1 approximately 5 were significantly longer than that of control, being 3.4, 8.4, 9.0, 6.1 and 8.8 days longer, respectively (P < 0.05). The MSTs of groups 2 approximately 5 were longer than that of group 1 (P < 0.05), but there was no statistical significance between groups 3 approximately 5 and 2. There was no statistical difference in peripheral blood count among groups 1 approximately 5. Histopathological examination of skin, liver, and gastrointestinal tract showed typical signs of aGVHD. Animals received immunosuppressive agents (MMF, CsA, MTX) showed the less severe signs.

CONCLUSIONSMMF markedly prolonged MST of allo-BMT recipients, delayed the onset of aGVHD signs. The prophylaxis effect of CsA + MTX with or without MMF was better than that of MMF alone, synergism between MMF of 10 or 30 mg.kg(-1).d(-1) and CsA + MTX was better than that of MMF of 60 mg.kg(-1).d(-1) and CsA + MTX.

Animals ; Bone Marrow Transplantation ; Cyclosporine ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Female ; Graft Survival ; drug effects ; Graft vs Host Disease ; pathology ; prevention & control ; Immunosuppressive Agents ; pharmacology ; Methotrexate ; pharmacology ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Time Factors ; Transplantation, Homologous