Objective To explore the effect of prevention of hemorrhagic cystitis (HC) after hematopoietic stem cell transplantation (HSCT) with hyperhydration, forced diuresis and alkalinizing plus infusion mesna. Methods 32 cases of patients receiving HSCT were included in this study. 2 cases of severe aplastic anemia (SAA) received total body irradiation (TBI)+cyclophosphamide(CTX)(TBI-CTX) regimen,and the remaining 30 patients were using the classic busulfan+CTX (BU+CTX) regimen. All patients were treated with mesna combined with hydration, forced diuresis and alkalization to prevent HC. Ganciclovir and acyclovir were used to prevent cytomegalovirus (CMV) and other viral infections and monitor CMV-IgM levels of the blood. Encourage patients to urinate every hour, testing urine pH value and the calculation of urine output, every 6 h review and testing of urine routine,central venous pressure (CVP), each of 8 h of serum electrolytes. Results Only 1 patient at 6 months after transplantation appeared delayed grade Ⅱ HC after hydration, alkalization, diuretic, hemostatic, anti-graft-versus-host disease (GVHD), and ganciclovir antiviral therapy. The HC patients cured at 35 d. The remaining patients did not suffer HC. Adverse effects such as acid-base balance disturbance did not appear clear. Conclusion Mesna joint hydration, forced diuresis and alkalization was effective and safe to prevent HC.

ObjectiveTo explore the efficacy and safety of bone marrow mesenchymal stem cells (BMSC)in treatment of aplastic anemia(AA). MethodsTwenty-two patients with aplastic anemia were enrolled with median age of 31 (12-70) years old,including 11 severe aplastic anemia (SAA),and 3 of whom were cyclosporine and anti-thymocyte globulin-resistant. BMSC were isolated from bone marrow of healthy donors and cultured.The third to fifth generation cells were administered intravenously in 1×106/kg to patients once or twice a week.After infusion,complete blood count,bone marrow aspiration,bone marrow biopsy,flow cytometry analysis of lymphocyte subsets of CD3+ CD4+ and CD3+ CD8+ and clinical symptoms were involved in outcome measurement.ResultsAll patients finished 16-time median (5-83 times) infusions of BMSC with 13-month median (2-33 months) treatment course and 23-month median (2-34 months) follow-up.The total response rate was 72.7 ％(16/22), including one patient with essential cure, 9 with remission, 3 with remarkably improvement and 3 with transfusion interval extension.Two of three front-line immunosuppressiveresistant SAA patients achieved remission. Ten of 14 patients recovered from inverted ratio of CD4+/C8+ cells after BMSC treatment. There were no treatment-related side effects observed in the course of treatment.ConclusionBMSC are effective and safe for the treatment of AA in our preliminary study and they deserve further research with larger-scale and long-term clinical trials.

BACKGROUND: Magnetic field can affect the growth and division of cancer cells both in vivo and in vitro, however, the effects on apoptosis of human liver cancer cells induced by magnetic field is still unclear.OBJECTIVE: To explore the inducing effect of extremely low fre quency (ELF) magnetic field on apoptosis of human liver cancer cell SK-HEP- 1.JESIGN: An open experiment with cells as the observational subjects.SETTING: Institute of Biotechnology, Shanghai Jiaotong University.MATERIALS: The experiment was carried out at the Institute of Biotechnology, Shanghai Jiaotong University from September 2004 to January 2005. The subject was human liver cancer cell line SK-HEP-1, purchased from cell bank of Chinese Academy of Science, Shanghai.METHODS: SK-HEP-1 cells were inoculated to T-flasks at the density of 2.0×107 cells L-1, and cultivated in the DMEM containing 0.1 volume fraction of heat-inactivated fetal bovine serum and 2 mmol/L L-glutamine. Exposure groups were exposed to 50 Hz, 20 mT magnetic field and the control groups were run concurrently under the same conditions with the exposed cultures but in a separate incubator which was free of magnetic field during 8-day culture process. The apoptosis of SK-HEP-1 cells were defined by DNA ladder assay, Hoechst 33258 staining and AO/EB staining respectively on day 8.MAIN OUTCOME MEASURES: ① DNA fragmentation pattern formation. ② The abnormal nucleus formation. ③ The percentage of apoptotic cells.RESULTS: ① Detection of internucleosomal DNA fragmentation by DNA ladder assay: After 8-day ELF magnetic field exposure, DNA fragmentation pattern was detected by DNA ladder assay, which was not observed in control groups (free of exposure). ② Fluorescence microscopy analysis of apoptosis by Hoechst 33258 staining: Hoechst 33258 staining was used to investigate the changes in the nucleus of cells, and many apoptotic bodies containing nuclear fragments were found in ELF magnetic field exposed cells, but just about none in untreated cells. At the same time, cytoplasmic shrinkage was observed in cells cultured under the exposure. And in some cells, even the cell membrane was unable to keep intact. ③ Fluorescence microscopy analysis of cell apoptosis by acridine orange/ethidium bromide (AO/EB) double staining: After ELF magnetic field exposure, the percentage of viable cells in exposure groups (9.2%) was rather low compared with the control groups (91.8%), and was accompanied with a high apoptotic rate (72.3%), while only 4.2% in control group. A large number of apoptotic cells were at the early stage of apoptosis. The rate of apoptotic cells (18.5%)after treated by magnetic field was higher than that of control group (4%). With the AO/EB double staining, control cells appeared to be round,intact and bright green while some of the exposed cells exhibited irregular cell morphology and condensed nucleus.CONCLUSION: Apoptosis of human liver cancer cell SK-HEP-1 could be induced by 50 Hz, 20 mT magnetic field in vitro.

Objective To investigate the changes in cytotoxicity of DC-CIK cells to human multiple myeloma RPMI 8226 cells before and after treatment with oridonin. Methods Normal human peripheral blood mononuclear cells were isolated and induced to obtain DC-CIK cells. Cytotoxicity of DC-CIK cells against RPMI 8226 cells which were treated by oridonin was analyzed by LDH releasing assay. The variation for expression of NKG2D ligands on RPMI 8226 cells were measured by flow cytometry. Results DC-CIK cells were successfully induced from the peripheral blood mononuclear cells. At the same effector to target ratio, oridonin obviously enhanced the cytotocixity of DC-CIK cells against RPMI 8226 cells (P<0.01). Flow cytometry showed the expression of NKG2D ligands ULBP1 of RPMI8226 cells was most significantly increased as the cells were treated by oridonin [(9.19 ± 1.85) vs. (15.47 ± 0.67), P<0.01]. Correlation analysis indicated that cytotocixity was positively correlated with changes in ULBP1. Conclusions Oridonin can improve the cytotoxicity of DC-CIK cells against RPMI 8226 cells, which may be related with the increased expressions of NKG2D ligands on the tumor cell surface.

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Objective:To investigate the influence of acrylamide ( ACR) on adult neurogenesis and expression of GSK 3βin mouse.Methods:Method Adult male Kunming mice were used and divided into two groups:control group and experimental poisoning groups,that were exposured to acrylamide by intraperitoneal injection.Brdu labeling and immunohistochemistry were used to investigate the proliferation of adult neural stem cells in the subgranular zone ( SGZ).BrdU/NeuN/GFAP triple labeling to investigate the survival and differentiation of newly generated cells.Detecting GSK3βexpression and distribution in Neuro-2a cells,the expression of GSK3βwas examined by using Western blot.Results:Compared with control mice ,lower number of BrdU-positive cells and less differentiated into neurons in ACR mice.Less neural stem cells survived ,but more glia cells were generated in the subgranular zone of acrylamide mice.Moreover,higher phosphorylated GSK 3β( Ser9 ) were detected in Neuro-2a cells and mouse dentate gyrus in ACR mice respectively .Conclusion:These results suggested that acrylamide inhibits neural stem cells proliferation and influences the survival and differentiation of newly generated cells.Acrylamide inhibits neurogenesis maybe through GSK 3βsignaling pathway.

Objective: To study the effects of extremely low frequency (ELF) magnetic field with fixed parameters on human liver cancer cells (SK-HEP-1) at different aspects. Methods: SK-HEP-1 cells were exposed to 50Hz, 20mT magnetic field during the whole culture process, and then proliferation activity, growth kinetics, metabolic profile and cell cycle were analyzed. Results: 50Hz, 20mT magnetic field inhibits the growth and metabolism of SK-HEP-1 cells, and hampers their mitotic division. Conclusion: 50Hz, 20mT magnetic field could be a potential therapy in the treatment of human malignant tumors.

Objective To summarize the therapeutic effects of combined hyperbaric oxygen on treatment of critical ill patients and the relevant problems to attention.Methods One hundred and forty-six critical ill patients, admitted to our hospital from January 2009 and December 2013, were chosen in our study;the relations of hyperbaric oxygen efficacy with different diseases and intervention times were analyzed.Methods and measures of safe and effective treatments for hyperbaric oxygen were summarized.Results The critical ill patients were treated with hyperbaric oxygen, with an effective rate of 73.97％ and a cure rate of 32.19％.The percentage of intervention time for cured patients within 30 days (73.34％) was obviously higher than that of more than 30 days (27.66％);and the hyperbaric oxygen times in patients with early treatment (less than 30 days) was obviously less than that in patients with delayed treatment (more than 30 days).There was no obvious difference between different diseases.Conclusions Hyperbaric oxygen therapy is effective for all kinds of critical ill patients.The cure rate is high and the treatment times are less when early intervention is applied.The patients with delayed intervention should insist on the long course of treatment so as to achieve satisfactory effect.And, it is the key to make the safe and effective treatments for patients with complicated diseases.

Objective To study the effect of a new immunomodulator composed of muramyl dipeptide(MDP) and anti-CD10monoclonal antibody(MDP-Ab)on the dendritic cells(DC)of children with acute leukemia. Methods DC was adopted to divide the children with acute lymphoblastic leukemia into 6 groups,including the control group,unconjugated anti-CD10alone,unconjugated MDP alone,MDP-Ab alone,lipopolysaccharide(LPS)alone and MDP-Ab + LPS.The immunophenotypes,the endocytosis interleukin-12(IL-12)were detected.The stimulation index of autologous lymphocytes was assayed by adopting 5-(and 6)-carboxyfluorescein diacetate,succinimidyl es-ter(CFSE)-staining method.The supernatants of DC and autologous lymphocytes were used to detect the level of in-terferon-γ(IFN-γ)by using enzyme-linked immunosorbent assay.Results (1)DC immunophenotype:The ex-pressions of human leukocyte antigen-DR(HLA-DR),mature molecule(CD83)and co-stimulatory molecules (CD80and CD86)were increased significantly upon DC triggered with MDP-Ab,compared with the control group,un-conjugated anti-CD10group,and unconjugated MDP group,but lower than those in LPS and combination of MDP-Ab with LPS(F=629.62,P=0.000).(2)The level of IL-12:a significant increase in IL-12 level was detected in MDP-Ab group,LPS group,and combination of MDP-Ab with LPS group,compared with the control group,uncon-jugated anti-CD10group,and unconjugated MDP group(F=857.87,P=0.000). There were significant differences among the first three groups.(3)Endocytosis assay:The uptake of DCs stimulated by unconjugated anti-CD10,un-conjugated MDP,MDP-Ab immunoconjugate,LPS or combination was lower than that of immature DC in the control group which was(81.3 ± 10.1)%.(4)Mixed lymphocyte reaction and IFN-γ level:DC,treated with MDP-Ab, LPS and combination,stimulated more CFSE positive cells and higher level of IFN-γ secretion than the control group and unconjugated anti-CD10group,unconjugated MDP group. The most significance was observed in combination of MDP-Ab with LPS(F=393.36,P=0.000;F=2 497.18,P=0.000).Conclusion It is concluded that MDP-Ab could promote the proliferation and maturation of DC derived from blood of children with acute leukemia.

glioma; U87 cell; U251 cell; MAGE-A1; MAGE-A3; DNA methylation; histone acetylation To detect the expressions of melanoma antigen genes MAGE-A1 and MAGE-A3 in glioma tissues and to explore their clinical significance. Methods: Seventy-eight surgically resected glioma specimens and 15 normal brain tissue samples from donors suffered traffic accidence were collected at the Department of Neurosurgery, the Fourth Hospital of Hebei Medical University between January 2006 and January 2010, and the mRNAexpressions of MAGE-A1 and MAGE-A3 in collected tissues were detected with RT-PCR; their associations with the overall survival of patients were also analyzed.The promoter methylation status of the two genes was observed with methylation specific PCR, and the relationship between the gene expressions and promoter methylation status was analyzed. The expressions of MAGE-A1 and MAGE-A3 genes in U251 and U87 glioma cell lines were detected by RT-PCR before and after the treatment with DNA methyltransferase inhibitor 5-aza-CdR and/or histone deacetylase inhibitor trichostatin A (TSA). Results:The positive expression rates of MAGE-A1 and MAGE-A3 genes in glioma tissues were 65.34% and 38.46%, respectively; however, the two genes were not detected in 15 cases of normal brain tissues.The 5-year overall survival of patients in MAGEA1 positive expression group was shorter than that of negative expression group (P<0.05). There was significant correlation between the mRNA expressions of two genes and their promoter methylation status (all P<0.01). There was no mRNA expressions of MAGEA1 and MAGE-A3 in U87 cells untreated with 5-Aza-CdR and TSA, but a small amount of MAGE-A1 mRNA and MAGE-A3 mRNA were detected in U251 cells. TSA alone could not activate the expression of MAGE-A1 and MAGE-A3 genes. 5-Aza-CdR alone or in combination with TSA could activate the expression of both genes, and the combined effect was better than that of single administration. Conclusion: There are different degrees of MAGE-A1 and-A3 expression in glioma tissues, and the expression of MAGE-A1 is a negative prognostic factor for glioma patients. DNApromoter methylation and histone acetylation are important mechanisms of the activation of MAGE-A1 and MAGE-A3 expression.