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 analyze the clinical features of renal involvement associated with Behcet's disease (BD) through 1 case and to make a review of the literature in order to early diagnose and cure in time,thus decrease misdiagnosis and mistreatment.Methods This is a retrospective study.The case was diagnosed with BD and the renal damage was confirmed by renal biopsy.The clinical features and histology features were analyzed.Results The presentation of renal disease was edema,proteinuria and microscopic hematuria.The clinical spectrum of renal BD showed a wide variation.Amyloidosis (AA type),GN (nephritis),and microscopic vascular disease were the main causes of renal BD.Patients with vascular involvement had a high risk of amyloidosis and amyloidosis was the most common cause of renal failure in BD.Conclusion Kidney is one of the organs that can alter the prognosis of the BD,so the screening for renal damage must be done for each patient with this disease.Routine urine analysis and measurement of serum creatinine level are needed for early diagnosis of renal BD.Immunosuppressive drugs can be useful in selected cases.

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 investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.

Public hospital reform pilots have been initiated recently and it is necessary to clarify some key issues. To this end, thispaper touches upon six fundamental issues, discussing the generality and value for the existence of public hospitals, differences between reforms of public hospital and those of state-owned enterprise, the public financing for and regulation on public hospitals, relationship with the private sector, as well as service and function positioning of public hospitals.

Objective To investigate the change patterns of operation time of laparoscopic appendectomy and its significance. Methods The clinical data of 105 consecutive patients with appendicitis who received laparoscopic appendectomy at the Southwest Hospital from January 2007 to March 2010 were retrospectively analyzed. Of the 105 patients, five were converted to open surgery, and they were excluded from this study.The changes in operation time of different surgeons were statistically analyzed to detect the change patterns of the learning curve in laparoscopic appendectomy. Results A hundred cases of laparoscopic appendectomy were successfully performed by three surgeons. The mean operation time was ( 87 ± 36 ) minutes ( range, 30-217 minutes). No surgical injury happened during the operation, and the blood loss was under 10 ml. The learning curve of operation time was presented as a sine curve with an oscillating decreasing trend. The primary two cycles end at an average of 9.6 cases, which could be used as the end point of the learning curve of laparoscopic appendectomy. Conclusions The learning curve of laparoscopic appendectomy shows a typical oscillating decreasing trend. The preliminary study ends when 9.6 cases of operation are completed.