Objective To optimize the condition of converting murine naive CD4+ CD25-T cells ( effector T cells,Teffs) to CD4-CD8-double negative regulatory T cells ( DN Tregs) in vitro.Methods Naive Teffs from C57BL/6 mouse were isolated with magnetic activated cell sorting( MACS)and co-cultured with DBA/2 mature dendritic cells (mDCs) with different doses of recombinant murine interleukin-2 (IL-2).The percentage of converted DN Tregs was examined by flow cytometry after 6 days.Purified DN Tregs were co-cultured with CFSE labeled Teffs.The proliferation rate of Teffs were evaluated by flow cytometry.Results Without IL-2,the percentage of CD4-CD8-T cells was 6.21％ ± 2.03％.With IL-2,the percentage was 14.77％ ± 2.15％ ( 25 ng/ml),21.29％ ± 2.68％ (50 ng/ml),43.45％ ±4.45％ (75 ng/ml),and 28.59％ ±3.05％ ( 100 ng/ml) respectively.The IL-2 concentration of 75 ng/ml signilicantly enhanced the conversion of Teffs to DN Tregs ( separately t =10.700,8.288,6.158,3.932,all P ＜ 0.05).Highly purified DN Trega significantly suppress the proliferation of Teffs in vitro.Conclusions Teffs are converted to DN Trega in vitro with the LPS-activated allogeneic mDCs and that 75 ng/ml of IL-2 is the optimal concentration for the conversion of Teffs to DN Trogs in vitro.

Objective:To study the effects of polysaccharide from Lycium barbarum (LBP) on cell apoptosis and bc1-2 gene expression in irradiated mice. Methods:Forty eight Kunming mice were divided into three groups (LBP group, radiation group and normal group). LBP chow was prepared through adding 0.8%LBP to normal chow and was supplied to LBP group. Normal chow was supplied to normal group and radiation group. LBP group and radiation group were exposed to whole-body 60Co ?-rays at the dose of 0.084 Gy/day for 6 w, five times a week and the total dose was 2.52Gy. Then the micronucleus frequency of polychromatic erythrocytes(MF),chromosome and sperm aberration frequency, caspase-3 mRNA expression, cell apoptosis and bcl-2 gene expression were detected. Results:LBP could significantly lower MF, chromosome and sperm aberration frequency, and cell apoptosis , and it could increase the proliferation activity of bone marrow cells and bc1-2 gene expression in irradiated mice and decrease caspase-3 mRNA expression.Conclusion:The radioprotective effect of LBP is related to regulation of cell apoptosis and bc1-2 gene expression.

Objective: To study the radioprotective effect of flavonoids from Ginkgo biloba leaves(GBF). Methods: Three water extracts of GBF were prepared (low dosage 10 mg/100 ml, medium dosage 20 mg/100 ml and high dosage 100 mg/100 ml) and orally administered to mice . After 10 d, the mice were exposed to 8.5Gy -rays. After another 10 d of oral administration, the survival rates were recorded in 30 d. In another experiment, six groups of mice (three GBF groups, radiation control, normal control and cyclophosphamide group) were arranged. The first three groups were orally administered with low, medium and high dosage of GBF respectively for 11d; the other three groups with distilled water. Then the three GBF groups and radiation group were exposed to 1.0Gy -rays. Then they were orally administered again in the following 7d . Micronucleated polychromatic erythrocytes in bone-marrow and sperms (AFS) in mice were observed on the 21st day after termination of oral administration. Proliferation rates of lymphocyte (PRL) were determined in the three GBF groups and normal control. Results: Low, medium and high dosage of GBF increased the survival rates by 31.7%, 25.3% and 26.5% respectively(P

ObjectiveTo study the effect on humoral immunity with combination of CsA and IL-2 fusion protein. MethodsForty C57/B6 mice were evenly divided into four groups,after receiving skin graft from DBA mouse.Mice in the experimental group was given CsA(30 mg/kg,ip) plus IL-2/Fc (1μg,ip) while the control group was only given CsA,each group was given HBV vaccine after skin graft surgery (2 μg,im),while blank group was only given vaccine after skin transplantation.The fourth group were left intact.Fourteen days later,the level of HBSAb,IL-4,IL-10,IFN-γ,IL-2 were measured with ELISA and IL-21and FoxP3 expression level and Tfh percentage of mixed lymphocytes detected. ResultsThe HBSAb level in experimental group is significantly higher than that in the control group and the survival time of skin graft is longer than that in the control group ( F =29.886,P =0.010 ; F =29.772,P =0.011).IL-2,IFN-γ are significantly higher than that in the control group( F =18.156,P =0.0020;F =90.042,P =0.003 ),but the Th2 cytokines such as IL-4,IL-10 are lower ( F =42.102,P =0.009 ; F =23.734,P =0.015 ).The expression level of both IL-21and FoxP3 are significantly higher than control group( F =9.784,P =0.048 ;F =27.883,P =0.012). ConclusionsCombination of CsA and IL-2 fusion protein can significantly enhance the immunogenicity of HBV vaccine and prolong graft survival time.It may be related to the higher expression level of IL-21and FoxP3.

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.