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.

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

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 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 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 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

Objective To investigate the clinical value of a new anvil inserting method for esophagogastrostomy or esophagojejunostomy during laparoscopic radical proximal gastrectomy or radical total gastrectomy for gastric cancer.Methods The clinical data of 21 patients with gastric cancer who received laparoscopic radical proximal gastrectomy or radical total gastrectomy at the Southwest Hospital from March 2010 to February 2011 were retrospectively analyzed.Five trocars were inserted through the abdominal wall of the patients.After perigastric lymphadenectomy and mobilization of esophagus,an incision was made on the esophagus above the tumor,and then the anvil with drawn wire attached was inserted into the esophagus.An endo-cutter was applied to cut the esophagus adjacent to the incision left the drawn wire untouched,and then the stem of the anvil was pulled out by the drawn wire for laparoscopic anastomosis. Results The operations were successfully accomplished under the laparoscope with no conversion to open surgery.Fifteen patients received laparoscopic radical total gastrectomy and 6 received laparoscopic radical proximal gastrectomy. The mean operation time,volume of blood loss,time to off-bed activity,passage of flatus and postoperative duration of hospital stay were (257 ± 38) minutes,( 119 ± 32) ml,(2.5 ± 0.5 ) days,( 3.7 ± 0.8 ) days and (7.5 ± 2.6) days,respectively.No perioperative mortality,anastomotic bleeding or anastomotic fistula was detected.One patient was complicated with pulmonary infection + pleural effusion and was cured by conservative treatment; 1 was complicated with anastomotic stenosis which was alleviated by gastroscopic balloon dilation; 1 was complicated by incisional infection and was cured by medical treatment after drainage.No cancer cells were detected at the anastomotic ring or resection margin of the specimen.There were 4 patients with well-differentiated adenoma,8 with moderate-differentiated adenoma and 9 with poor-differentiated mucinous adenoma.There were 5 patients in stage Ⅰ,10 in stage Ⅱ and 6 in stage Ⅲ (UICC staging).Twenty-one patients were followed up for a mean period of (11 ±4) months (range,6-17 months ),no tumor recurrence or metastasis was detected. Conclusions The new technique for anvil insertion is safe,effective and easy for manipulation and learn.It offers a new approach for laparoscopic digestive tract reconstruction.

Objective To summarize the experiences in laparoscopic inguinal hernia repairing for adult patients. Methods Clinical data of 512 hernia cases admitted in our center from March 2007 to Sep 2010 were retrospectively analyzed.There were 437 cases of single-sided hernia,including 281 indirect inguinal hernia,86 direct inguinal hernia,15 femoral hernia,16 combined inguinal hernia and 39 recurrent hernia.There were also 75 cases of double-sided inguinal hernia,including 3 recurrent hernia.There were 41 acute incarcerated hernia cases.The average postoperative follow up time was(29 ± 12) months. Results 507 cases underwent successful laparoscopic repair,and 5 cases were converted to open procedure.There were 238 TAPP and 269 TEP in laparoscopic operations.The average operative time for TAPP was (69 ±19) min,and (58 ±15) min for TEP.The average length of postoperative stay was (5.0 ± 1.5) days.The percentage of resuming normal activity after 2 weeks and 4 weeks were 95.7％ (485/507) and 99.0％(502/507).The most common postoperative complications were seroma (9.7％,49/507),transient paresthesia (4.1％,21/507) and chronic pain (0.8％,4/507).The recurrence rate was 0.6％ (3/507).Conclusions Laparoscopic repair of inguinal hernia has the advantage of less trauma,faster recovery,and lower recurrence rate.

Objective To summarize the experience in application of da Vinci robotic surgical system in radical resection of rectal cancer,and investigate the proper position of trocars and operative techniques.Methods The clinical data of 13 patients who received radical resection of rectal cancer accomplished by the da Vinci robotic surgical system at the Southwest Hospital from February 2010 to February 2011 were retrospectively analyzed.The patients were in lithotomy position and received combined intravenous anesthesia.Five or 4 trocars were used.Miles procedures were performed on patients with lower tumor position,and the other patients received Dixon procedure.Results The operation was successfully performed on all patients.Five trocars were selected for the first 3 patients,and 4 trocars for the other 10 patients.Nine Dixon procedures and 4 Miles procedures were selected.The mean operation time was 217.3 minutes (range,160-260 minutes).The mean operative blood loss was 53.3 ml (range,40-70 ml) in Dixon procedure and 120.0 ml (range,90-130 ml) in the Miles procedure,and no blood transfusion was needed.The mean number of lymph nodes dissected was 13.9 (range,8-21 ),and the time to bowel movement was 3.2 days (range,2-5 days).Two patients were complicated with pulmonary infection,1 with urinary tract infection,and they were cured by antimicrobial therapy.No other morbidity or mortality was found.The results of postoperative pathological examination showed that there were no residual cancer cells at the resection margin,and the distance between the resection margin and the tumor was 6.3 cm (range,3-10 cm).There were 1 patient in stage Ⅰ,5 in stage Ⅱ and 7 in stage Ⅲ.The mean time of follow-up was 5.9 months (range,3-12 months),and no recurrence or metastasis was found during follow-up.ConclusionsRadical resection of rectal cancer with da Vinci robotic surgical system utilizing 4 trocars has the advantages of minimally invasive surgery with fast recovery as well as the ease of dissection afforded by the surgical robot.