The interaction between the immune system and nervous sytem affcts nervous function.There is a close relationship between the episode of epilepsy and immunity,the molecular basis of which is the network of cytokines.In order to quest new methods of contolling epilepsy by regulating the network of cytokines,the roles of several cytokines in the ocurrence and onset of epilepsy were reviewed in this article.

Objective:To investigate the role of blockade of CD40 ligand-CD40 costimulatory pathway by anti-CD40L mAb on T lymphocytes typing and cytokines, and provide the experimental clues of inducing donor-specific T cell anergy.Methods:To monitor primary MLR, C57BL/6 H-2b spleen T cells were isolated as responder cells, and BALB/C H-2d spleen cell as stimulator cells. Anti-CD40L mAb was added to primary MLR cultures(termed anti-CD40L mAb group), and in control group(no anti-CD40L mAb). Incubated for 7 days, the cells responsiveness rates were detected by 3 H-TdR methods at the indicated time points, and supernatants were assayed for IFN-?,IL-2,IL-4,IL-10 cytokines levels by commercial ELISA kits. Day 5 MLR-cultured cells were assessed for the expressions of CD4, CD8, CD25, CD69, CD40L and CD45RA with flow cytometry(FCM).Cells proliferation for 5 days and cytokines in secondary MLR were assayed according above-mentioned method.Results:Blockade of the CD40-CD40L pathway by anti-CD40L mAb can induce the hyporesponsiveness to alloantigen in primary and secondary MLR culture. In primary MLR culture, the expressions of CD4 +T and CD8 +T cells and CD4 +CD25 +T, CD4 +CD69 +T, CD4 +CD40L +T, CD8 +CD25 +T and CD8 +CD69 +T cells in anti-CD40L mAb group were lower than that in control group ( P 0.05).The expression of CD4 +CD45RA + T in anti-CD40L mAb-cultured group was higher than that in control group( P

0 05). The level of IL-12 in PB serum was higher than that of PB serum( P

AIM: The expression of the mouse ? 2 microglobulin (? 2 m) in NIH3T3 cells transfected with the mouse ? 2m sense and antisense RNA was detected to clarify the effect of mouse ? 2m sense and antisense RNA on the expression of MHC classⅠgene. METHODS: The mouse sense and antisense RNA, pcDNA3-? 2mSN and pcDNA3-? 2mAN, were constructed and were transfected into NIH3T3 cells by lipofectamine. RT-PCR and Western blot were used to detect the expression of ? 2m in those cells. RESULTS: The expression of the mouse ? 2m in the cells transfected with pcDNA3-? 2mSN was increased, while it was decreased in those cells transfected with pcDNA3-? 2mAN. CONCLUSION: pcDNA3 -? 2mAN can downregulate the expression of the ? 2m in NIH3T3 cells.

[Objective]To investigate the effects of mesenchymal stem cells (MSC) feeder layer, culture sere and freeze-thaw lysates on expansion and differentiation of cord blood CD34~+ cells in vitro. [Methods] MSC were isolated from human bone marrow and cultured until the third passage. Sera were obtained from the cultured MSC, and freeze-thaw lysates were obtained by repeated freeze-thaw procedures. Cord blood CD34~+ cells were isolated by magnetic cell separation system, and were co-cultured with the MSC feeder layer, culture sera, freeze-thaw lysates and hematopeietic growth factors (HGFs), respectively. The nucleated cells, CD34~+ cells, CD34~+CD38~- cells, CD41~+ cells and CD3~+ cells in the above culture system were detected by flow cytometry on day 6 and day 12. [Results] ①MSC feeder layer had a strong effect on nucleated cells, CD34~+,CD34~+CD38~- cells expansion. The MSC sera and freeze-thaw lysates had similar effect on cell expansion, but the effect was weaker than that of feeder layer (P<0.05). ② Both MSC sera and feeder layer inhibited cord blood CD34~+ cells differentiation toward CD3~+ cells or CD19~+ cells, and no significant differences were found between these two groups (P>0.05). ③ Both MSC sera and feeder layer promoted cord blood CD34~+ cells differentiation toward CD41~+ cells, and the effect was stronger in the feeder layer than that of the sera (P<0.05). ④ Freeze-thaw lysates had no effect on cell expansion and differentiation, and were similar with that of HGFs (P>0.05). [Conclusions] The MSC sera have positive effects on expansion of cord blood CD34~+ and CD34~+CD38~- cells, moreover they have the ability of promoting cord blood CD34~+ cells differentiation toward CD41~+ cells.

AIM:To investigate the potential of hematopoietic stem cells(HSCs)derived from mouse embryonic stem cells(ESCs)to reconstruct hematopoiesis in vivo.METHODS:Using a three-step method,a mice embryonic stem cell line,E14.1 was induced into hematopoietic stem cells.The cell markers with CD34+/Sca-1+ were identified by flow cytometry analysis,then HSCs(1?109 cells/L)from third-step were injected into SCID mice for observing teratoma formation.To validate function of HSCs,colonogenic cell assay was conducted and the hematopoiesis in lethally irradiated mice was reconstituted.RESULTS:The method of three-step differentiation,combined to use more hematopoietic stimulating factor promoted the E14.1 cell differentiation into HSCs with highest percent of CD34+/Sca-1+ cells(as high as 58.64%?4.20%)with more CFU-E,CFU-GM and CFU-GEMM populations.The cells showed the character of hematopoietic progenitors by Wright-Giemsa staining.Positive selected CD34+/Sca-1+ cells by magnetic sorting from third-step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 d after transplantation,with 71.4%(5/7)successful engraftment rate.Three recipients showed that the cell population of the peripheral blood leukocytes,red blood cells and hemoglobin approached to normal index at 40 d after transplantation,but followed relative slow renew in platelet count.Survival rate in transplant group was 43%,compared to 100% mortality in control mice.Karyotyping assays confirmed the female mice with XY.CONCLUSION:The three-step differentiation and the culture conditions described here support the differentiation of mouse ESCs into HSCs.HSCs derived from mouse ESCs can reconstruct hematopoiesis.

OBJECTIVE To investigate the characteristic and prognosis of nosocomial infection in children patients with malignances during deficiency stage of neutropenia. METHODS The clinical characteristic of infections and efficiency of antibiotics were analyzed retrospectively in 174 malignances children with nosocomial infection from Jan 2000 to Jun 2005. RESULTS The incidence rate of nosocomial infection in children patients with malignances during deficiency stage of neutropenia was 67.4%.Nosocomial infection was occurred mainly in the respiratory tract,oral cavity,blood,skin,and intestinal tract of patients.The main pathogens were bacteria(86.8%),and fungal infection was 13.2%.The Gram-negative bacilli were relatively sensitive to imipenem,amikacin,ceftazidine,piperacillin/tazobactam,and ticarcillin/clavulanic acid.The Gram positive cocci were sensitive to vancomycin,imipenem,meropenem,ampicillin/sulbactam,ciprofloxcin and clindamycin.The death rate due to nosocomial infection was 44.4%. CONCLUSIONS There is higher incidence of nosocomial infection in children patients with malignances during the neutropenia stage.Good nursing,intestines disinfection,the usage of granulocyte colony-stimulating factor,a reasonable usage of antibiotics and preventing the fungal infection are good to control nosocomial infection in children malignances.

AIM: In order to replace mouse embryonic fibroblast cells by human embryonic fibroblast cells to support the growth of human embryonic stem cells, the effects of human/mouse embryonic fibroblast cells on the growth of human embryonic stem cells were compared. METHODS: Both mouse and human embryonic fibroblast cells were used as feeder layer to support human embryonic stem cells. The proliferation and differentiation of human embryonic cells were observed. RESULTS: Combined use of human leukemia inhibitory factor with human/mouse feeder layer cells would support growth and proliferation of human embryonic stem cells and kept in undifferentiated condition. There was no difference between human/mouse cell. CONCLUSION: Human embryonic fibroblast cells can be used to support proliferation of human embryonic stem cells, eliminating the influence of foreign protein.

AIM: To explore inductive methods of hematopoietic stem cell (HSC) from embryonic stem cells (ESC) in vitro. METHODS: Using mice E14 line, the first step was the primary differentiation in which the ESC form embryoid bodies (EB) in methylcellulose-based cultures with SCF and VEGF. The second step involved the plating of cells originating from the EB into three different system of cultures containing SCF, VEGF,IL-3, IL-6 and EPO for HSC. And identifying HSC by flow cytometry analysis, colonogenic cells assay and Wright-Giemsa stain were also used. RESULTS: By two-step differentiating, it showed that HSC differentiated slowly in methylcellulose medium, percent age of CD34+/Sca-1+ cells slight increased about(31.5?4.7)% after day 14 induction. However, EBs were induced after 10 days to fast differentiate for HSC with more cells population by coculture on bone marrow stromal cells feeder. Flow cytometry analysis showed that percentages of CD34+/Sca-1+ cells might reached to (47.8?6.3)%. The more optimistic system of differentiation was bone marrow stromal cells feeder (BMSCF) in combination with supernatants of stromal cells from mice fetal liver(SSCFL), it significant supported differentiation of ESC into HSC with higher percent (53.6?7.2)%. Colonogenic cell assay and Wright-Giemsa stain confirmed that it possessed character of hematopoietic progenitors. CONCLUSION: Using methods of two-step differentiation, mice ESC were induced to differentiate into HSC by coculturing with BMSCF and SSCFL in combination of SCF,VEGF,IL-3,IL-6 and EPO.

AIM: To investigate the sequence expression o f CD158 molecule after tacrolimus (FK506), mycophenolate mofetil (MMF) combined with methylprednisone (MP) treatment for refractory chronic graft-versus-host di seases (cGVHD). METHODS: The efficacy and the side effect were observed in 6 chi ld patients with extensive cGVHD after allogeneic hematopoietic stem cell transp lantation treated with the combination of FK506, MMF and MP, meanwhile the chang es of the CD158 expressions on T lymphocytes and NK cells in peripheral blood be fore and after treatment were observed. RESULTS: The expression of CD4+CD158a+ and CD4+CD158b+ were very low before and after transplantation and treatment, there was no stati stical significance. The expression of CD3+CD158b+ and CD3+CD8+CD158b + were 4.97%?2.36% and 4.58%?2.90% respectively in five patients with acut e GVHD, and there was statistical significance compared with that of before-tran splantation (P