Adolescent ; Bone Marrow Transplantation ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Thalassemia ; surgery ; Transplantation Chimera

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective To investigate the mechanism of protective effects of resveratrol on tissueengineered cartilage.Methods The chondrogenesis of alginate-encapsulated bone marrow mesenchymal stem cells (BMSCs) were evaluated by toluidine blue staining and immunostain.The morphology of BMSCs-derived chondrocytes cultured on chitosan-gelatin scaffolds (CGS) was evaluated by scanning electron microscope and laser confocal microscope.When these cells on CGS were pre-stimulated with interleukin-1β (IL-1β) or cotreated with IL-1β and resveratrol in the absence and presence of specific β1-integrin blocking antibody,collagen type Ⅱ,aggrecan,matrix metalloproteinase-13 (MMP-13) expression,and the translocation of nuclear factor kappaB (NF-κB) were analyzed by Western blotting.ANOVA was used for statistical analysis.Results Alginate bead culture plus conditional medium together could induce the cartilage-specific collagen type Ⅱ,aggrecan expression and extracellular matrix accumulation in differentiated chondrocytes.CGS supported differentiated cell attachment,proliferation,and migration.When those cells cultured on CGS were stimulated with IL-1β alone,collagen type Ⅱ and aggrecan expression was inhibited.However,MMP-13 expression increased.By Western blotting semi-quantitative analysis,the expression level of cartilage-specific collagen type Ⅱ of the control group was 0.484±0.006; the expression level of resveratrol intervention group was 0.474±0.014.The difference between these two groups was not statistically significant (P＞0.05).The expression level of the IL-1β intervention group reduced to 0.155±0.009,which was statistically significant different from the above two groups(P＜0.05).Resveratrol could antagonist the negative effect of IL-1β,and increase collagen type Ⅱ to 0.468±0.014,the difference between these two was statistically significant (P＜0.05),and no significant difference when compared to the control group (P＞0.05).Specific β1-integrin blocking antibody could abrogate these effects of resveratrol,decrease collagen Ⅱ expression to 0.169±0.011,the difference was significant (P＜0.05),but there was no difference when compared to the IL-1β group (P＞0.05).Aggrecan semi-quantitative expression has the same trend in the expression of type Ⅱ collagen while the expression of MMP-13,NF-κB had the reversal trend.These indicated that the resveratrol reversed the catabolic effects by reducing the nuclear translocation of NF-κB.Specific β1-integrin blocking antibody abrogated these effects of resveratrol.Conclusion Resveratrol,by regulating β1-integrin,acts as a NF-κB nuclear trans-location inhibitor to protect tissue-engineered cartilage.

Objective To investigate the anti tumor effects of cytotoxic T lymphocytes induced by tumor antigen specific dendritic cells. Methods Soluble tumor antigen was prepared by four freeze thaw cycles of gastric tumor cell line SCG7901. Anti gastric tumor vaccine was acquired by co incubation of tumor antigen and mouse bone marrow derived dendritic cells and cultured with granulocyte/macrophage colony stimulating factor and interleukin 4. Antigen specific cytotoxic T lymphocytes were induced by this tumor vaccine from the spleen and were applied to tumor bearing nude mice. Results Tumor growth was significantly inhibited and the apoptosis of tumor cells was promoted extensively. Conclusions Antigen specific dendritic cell tumor vaccine may play an important role in future immunotherapy of gastric cancer.

OBJECTIVE To observe the protective effect and underlying mechanism of total ginsenosides (TG) on bleomycin-induced pulmonary fibrosis. METHODS Intratracheal instillation of bleomycin 5 mg · kg-1 was conducted to establish a pulmonary fibrosis mouse model. Kunming mice(1/2 males and 1/2 females)were randomly divided into sham-operation(Sham),model,total ginsen?osides 40,80 and 160 mg·kg-1 and prednisone acetate(5 mg·kg-1) groups. After 28 d administration,the histopathological changes in the lung were analyzed by hematoxylin eosin(HE)and Masson staining. The exprssion of alpha smooth muscle actin(α-SMA)in the lung was detected by real-time PCR. The content of hydroxyproline(HYP)and glutathione(GSH),level of total antioxidant capacity(T-AOC)and hydroxy radical(·OH),activity of myeloperoxidase(MPO)and nitric oxide synthase(NOS)in the lung were detected by corresponding kits. RESULTS Compared with the sham group,the pulmonary indexes in model group were significantly increased(P<0.01),alveolitis and pulmonary fibrosis were obvious. The mRNA expression ofα-SMA,content of HYP and · OH,activity of MPO and NOS were increased(P<0.05),but the content of GSH and T-AOC in model group was decreased(P<0.05). Compared with model group,the pulmonary indexes in TG 80 and 160 mg · kg-1 and prednisone acetate 5 mg · kg-1 groups were reduced(P<0.05),and the degree of alveolitis and pulmonary fibrosis was mitigated. The mRNA expression ofα-SMA,content of HYP and · OH, the activity of MPO and NO were decreased (P<0.05),while the content of GSH and T-AOC was increased(P<0.05). CONCLUSION TG can improve the degree of mice pulmonary fibrosis induced by bleomycin. The mechanism may be related to the increased antioxidant capacity of organisms.

Objective To investigate the relationship between CYP2J2*7 mutation(G-76T) and coronary heart disease (CHD) in Chinese Hanpopulation and to study the effects of CYP2J2 geneover-expressionon the proliferation and migrationof aortic smooth muscle cells of ApoE-/- mice. Methods CYP2J2*7 genotype was detectedin 500 patients with CHD and 478 controlsubjects by the Polymerase Chain Reaction-Restriction Frag-ment Length Polymorphism (PCR-RFLP). Culturedaortic smooth muscle cells of ApoE-/- mice were divided into control group, sham transfectiongroup and CYP2J2 over-expression group. Cell proliferation and migration were investigated after CYP2J2 over-expressionby MTS and Transwell assay. Results The frequency of CYP2J2*7 in CHD group was significantly higher than that incontrol group (10.00% vs. 6.49%, P = 0.046). Same is the case in female cases(P = 0.026). Compared with these of aortic smooth muscle cells incontrol group and sham trans-fectiongroup, the cell proliferation in 24, 48, 72 h, and the cell migration in 48 h after CYP2J2 over-expression in CYP2J2 group were significantly suppressed. Conclusions CYP2J2*7 mutation might increase the risk of CHD in Chinese Han population. CYP2J2 over-expression can suppress the proliferation and migration of aortic smooth muscle cells and CYP2J2 might have the effect of anti-atherosclerosis.

Objective To genotype Candida strains by random amplified polymorphic DNA(RAPD)technique,and to analyse the analogy of different Candida strains based on similarity coefficient.Methods Candida strains were isolated from vaginal secretion taken from patients with vulvovaginal can-didiasis by Sabouraud' s dextrose agar,C.albicans was differentiated from non-C.albicans strains by germ tube test,chlamydospore test and CHROMagar-Candida,and further identified by API20c Aux test Kits.The genomic DNA of45Candida isolates,including Candida albicans,Candida glabrata,Candida krusei,Candida parapsilosis and Candida tropicalis were amplified by RAPD,and DNA polymorphisms of inter-species and intraspecies were evaluated by using the DNA pattern-clustering of5primers.Results Thirty strains of C.albicans could be classified into7groups,and further divided into19subgroups,while15strains of non-C.albicans could be identified to species level.C.albicans isolates were related to each other with the similarity coefficients of more than90%,while different Candida species were connected to each other with the similarity coefficients of80%~90%.Conclusion C.albicans,which is the predominant pathogen of vulvovaginal candidiasis,could be classified into different genotypes by use of RAPD method.

Objective To investigate the expression and clinical significance of tumor infiltrating dendritic cells(TIDCs) within gastric tumor tissues.Methods Immunohistochemistry(IHC),in-situ hybridization(ISH) and flow cytometry were applied to detect the expression of S100,CD83 mRNA and CD83 on DCs in 45 gastric adenocarcinoma tissues.The co-relationship of the S100 and CD83 expression with clinical(pathological) features was analyzed.Results IHC showed that S100 expression was unevenly distributed(within) 42 cancer tissue and CD83 was only expressed in tumor-adjacent tissue and normal tissue.ISH showed that CD83 mRNA was limitedly expressed within 7 samples.S100 expression had no significant(correlation) with clinical pathological features,while CD83 reversely correlated with TNM stages.Detected by flow cytometry,CD83 was expressed in low level in all 45 gastric cancer tissues and negative correlations were found with lymph node metastasis and TNM stages of gastric cancer(r=-0.879,P

Child ; Contracture ; congenital ; therapy ; Humans ; Male ; Neck Muscles ; pathology ; Torticollis ; congenital

Objective To investigate the feasibility of laparoscopic-assisted transanal pull-through resection and anastomosis in the treatment of ultra-low rectal cancer.Methods From November 2005 to December 2006,21 patients with ultra-low rectal cancer had undergone laparoscopic-assisted transanal pull-through resection and anastomosis in Southwest Hospital.The perioperative condition,postoperative complications and the result of follow-up were retrospectively analyzed.Results The operation was successfully performed on all the patients.The mean operation time and postoperative hospital stay were(216±25)minutes(170-260 minutes)and(9.4±1.0)days(7-11 days),respectively.The time needed for the recovery of gastrointestina]function was(65±14)hours(38-88 hours).The mean perioperative blood loss was(140±49)ml(80-250 ml).All the patients were followed up for(22±4)months(15-28 months),and no anastomotic bleeding or fistula was observed.Six patients developed mild to moderate anastomotic striclure,1 local recurrence and 1 liver metastasis.Conclusions Laparoscopic-assisted transanal pull-through resection and anastomosis for ultra-low rectal cancer is safe and feasible,and the short-term effect is satisfactory.