Objective To explore the CD4 + CD25 + Foxp3 + regulatory T cell percentage and plasma levels of soluble CD25 molecules in peripheral blood of septic patients and their clinical value through prospective study. Method A total of 37 septic patients and 15 non-infectious SIRS patients, who conformed to the criteria of SIRS and sepsis which proposed by American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference ( ACCP/SCCM ) in 1997, were collected in ICU of Ruijin Hospital ( Shanghai Jiaotong University) from February 2009 to February 2010. Twenty-four health people were from Medical Center of Ruijin Hospital, who were excluded infection and (or) autoimmune diseases. There were 26 male and 11 female in sepsis group, average age ( 61.67 ± 11.87 ) years old; 8 male and 7 female in SIRS group, average age (67.06 ± 12.57)years old; 14 male and 10 female in health control, average age (56.54 ± 6.37 )years old. All selected patrents were excluded the autoimmune diseases and (or) patients within recent (30 days) had used or now used immunosuppressive agents. We therefore measured the Treg cell percentage in peripheral blood by Flow Cytometry and the plasma levels of IL-2sRa, IL-4, IFN-γ by ELISA. The data were analyzed by analysis of variance or nonparametric Kruskal-Wallis H test. Results ① The percentage of CD4 + CD25 + Foxp3 + regulatory T cells among septic patients, SIRS patients, and control group was: ( 66.82 ± 21.79 ) %, ( 51.79 ± 21.79 ) %, ( 56.45 ± 10. 68 ) %, respectively. septic patients showed the highest percentages of CD4 + CD25 + Foxp3 + regulatory T cell among CD4 + CD25 + T cells(P ＜ 0.05 ). ② The plasma levels of soluble CD25 in septic patients (425. 619 ± 270.12 ) were significantly higher than SIRS patients (381. 664 ± 189.83) and the control group ( 164. 1 32 ± 56.37 ) ( P ＜ 0.05 ). ③ The correlation analysis between the concentration of soluble CD25 molecules in plasma and the ratio of CD4 + CD25 + Foxp3 + regulatory T cells to CD4 + CD25 + T cells showed Spearman correlation coefficient =0.390, P = 0.003 ( P ＜ 0.05 ). Conclusion: the expression of natural regulatory T cells characteristically increased in septic patients. And the levels of soluble CD25 in peripheral blood were related to the percentages of natural regulatory T cells, which simplified the assessment of the immune status in Septic patients.

Objective To explore how to isolate and culture human primordial germ cells(PGCs).Methods Human embryonic fibroblasts(HEFs) isolated from embryos at 2 to 3 months of age received a radiation of 60Co ?-ray and then served as feeder layer cells.Cells collected from trypsinized human embryonic gonadal ridges and dorsal mesenteries(5 to 9 weeks post-fertilization) were cultured on HEF feeder layers in the medium with recombinant human leukemia inhibitory factor(LIF),recombinant human basic fibroblast growth factor(bFGF) and forskolin.Histochemical and immunocytochemical assays were used to identify the stage specific embryonic antigen-1and 4(SSEA-1/4) of the cultured cells.Reverse transcription polymerase chain reaction(RT-PCR) was used to identify the expressions of specific genes,including Oct-4,Ifitm-3,stella,Mvh and DAZL.Embryonic bodies were cultured as while.Results The collected cells were growing on the feeder layer and formed a typical nestlike multicellular colonies.The cells showed diploid chromosome in karyotype analysis and expressed SSEA-1/4 and specific genes,Oct-4,Ifitm-3,stella,Mvh and DAZL,indicating that the cells were PGCs.When cultured in hanging drop,the PGCs developed into embryonic bodies,showing multipotential ability.Conclusion Human PGCs can be isolated from genital ridges and dorsal mesenteries of embryos and cultured in vitro on HEF feeder layer.The formation of the embryonic stem cell colony can be observed,and the cells cultured by this method is confirmed to be embryonic germ cells.

Objective To investigate the value of critical scoring system in the treatment of multiple organ dysfunction after exposure to poisoning by analyzing 3 cases of industrial organic fluorine gas poisoning.Methods Clinical data of symptoms,signs,treatments,outcome and the changes in the scores as APACHE Ⅱ,SOFA,MODS were collected,the differences among the patients were compared and the relevance was analyzed.Results The first APACHE Ⅱ was 19-26.Scores of case 1 in the three scoring systems constantly increased and different components were found,with the suggestion of sequential organ dysfunction.Other two patients' scores decreased and survived without complications.Conclusiorn Multiple critical scoring systems can be used to evaluate the outcome of multiple organ dysfunction due to organic fluorine gas poisoning.System evaluation and individualized treatment are both important.More studies can help to set up a special critical scoring system.