Objective To investigate the mechanisms of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) combined with Triptolide in inducing the apoptosis of pancreatic cancer cells.Methods (1) The pancreatic cancer cells (MiaPaca-2 cells) were divided into 4 groups:blank control group (no drugs were added),TRAIL + Triptolide-group (only TRAIL was added),TRAIL-Triptolide + group (only Triptolide was added) and TRAIL+ Triptolide+ group (TRAIL and Triptolide were added).The vitality of cells in all the 4 groups was assessed by CCK-8.The expressions of poly ADP-ribose polymerase (PARP),cysteinyl aspartate specific proteinase-3 (Caspase-3) and Caspase-8 were detected by Western blot.The vitality of cells was detected by CCK-8 and the vitality of Caspase-8 was detected by Caspase-Glo assays after adding Z-IETD-FMK,a specific inhibitor of Caspase-8.The expressions of myeloid cell leukemia-1 (Mcl-1),Bcl-xL and Bcl-2 were detected by Western blot.(2) The MiaPaca-2 cells were divided into 8 groups:①TRAIL-Mcl-1 siRNA-group (no TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL+ Mcl-1 siRNA-group (TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL-Mcl-1 siRNA + group (TRAIL was not added and Mcl-1 siRNA cells were transfected)and TRAIL+ Mcl-1 siRNA+ group (TRAIL was added and Mcl-1 siRNA cells were transfected).②TRAIL-Bcl-xL siRNA-group (TRAIL was not added and Bcl-xL siRNA was not transfected),TRAIL+ Bcl-xL siRNA-group (TRAIL was added and Bcl-xL siRNA was not transfected),TRAIL-Bcl-xL siRNA + group (TRAIL was not added and Bcl-xL siRNA was transfected) and TRAIL+ Bcl-xL siRNA+ group (TRAIL was added and Bcl-xL siRNA was transfected).The vitality of the cells in all the groups was detected by CCK-8.The expressions of Caspase-3 and Caspase-8 protein were detected by Western blot.The measurement data with normal distribution were presented as (x) ± s.The comparison among groups was done by ANOVA,and the pairwise comparison was done by LSD-t test.Results (1) The vitalities of MiaPaca-2 cells in the blank control group,TRAIL + Triptolide-group,TRAIL-Triptolide + group and TRAIL + Triptolide + group were 100.0％ ± 1.1％,81.2％ ± 2.3％,78.6％ ± 3.6％and 40.1％ ± 2.5 ％,and the relative expressions of PARP protein were 0.510 ± 0.028,0.720 ±0.072,1.250 ±0.023 and 2.560 ± 0.220,the relative expressions of Caspase-3 were 0.080 ± 0.004,0.080 ± 0.003,0.110 ±0.005 and 2.720 ± 0.003,and the relative expressions of Caspase-8 were 0.070 ± 0.003,0.080 ± 0.005,0.120 ±0.003 and 0.990 ± 0.006,with significant differences among the 4 groups (F =203.607,1 457.785,332 421.900,35 437.218,P ＜ 0.05).The vitality of M iaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and the TRAIL-Triptolide + group (t =34.583,355.936,36.271,P ＜ 0.05).The relative expression of PARP protein of MiaPaca-2 cells in the TRAIL+ Triptolide + group was significantly different from those in the blank control group,TRAIL+ Triptolidegroup and TRAIL-Triptolide + group (t =591.784,63.739,2 268.987,P ＜ 0.05).The relative expression of Caspase-3 protein of the MiaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and theTRAIL-Triptolide + group (t =3 266.153,9 145.228,1 738.713,P ＜0.05).The relative expression of Caspase-8 protein of the MiaPaca-2 cells in the TRAIL+ Triptolide +group was significantly different from those in the blank control group,the TRAIL+ Triptolide-group and the TRAIL-Triptolide + group (t =663.953,l 432.878,327.584,P ＜ 0.05).The vitality of caspase-8 in the TRAIL+ Triptolide+ group was 711.0％ ± 5.1％ before adding Z-IETD-FMK,and then the vitality of MiaPaca-2 cells and caspase-8 changed to 70.0％ ± 4.8％ and 73.0％ ± 2.4％,with significant differences (t =17.956,55.027,P ＜ 0.05).The relative expressions of Mcl-1 protein in the blank control group,the TRAIL + Triptolidegroup,the TRAIL Triptolide + group and the TRAIL + Triptolide + group were 1.68 ± 0.22,2.08 ± 0.11,0.73 ±0.15 and 0.58 ± 0.18,the relative expressions of Bcl-xL protein were 0.65 ± 0.03,0.47 ± 0.03,0.32 ± 0.03and 0.26 ±0.05,the relative expressions of Bcl-2 protein were 0.65 ± 0.03,0.67 ± 0.03,0.62 ± 0.05 and 0.67 ± 0.03,with significant difference among the 4 groups (F =55.178,88.683,3.411,P ＜ 0.05).The relative expressions of Mcl-1 protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL+ Triptolide +group were significantly different from those of the blank control group (t =23.506,47.631,P ＜ 0.05) and the TRAIL + Triptolide-group (t =58.457,37.115,P ＜ 0.05).The relative expressions of Bcl-xL protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL + Triptolide + group were significantly different from those of the blank control group (t =38.105,42.219,P ＜ 0.05) and the TRAIL + Triptolide-group (t =32.476,15.814,P ＜ 0.05).The relative expressions of Bcl-2 protein in the TRAIL-Triptolide + group and the TRAIL+ Triptolide + group were not significantly different from those of the blank control group (t =4.724,1.732,P > 0.05) and the TRAIL + Triptolide-group (t =3.464,0.000,P ＞ 0.05).(2) The vitalities of MiaPaca-2 cells of the TRAIL-Mcl-1 siRNA-group,TRAIL + Mcl-1 siRNA-group,the TRAIL-Mcl-1 siRNA + group and the TRAIL + Mcl-1 siRNA + group were 100.0％ ± 2.2％,79.3％ ± 1.8％,71.2％ ± 3.2％ and 37.3％ ± 5.4％,the relative expressions of Caspase-8 protein were 0.100 ± 0.003,0.100 ± 0.005,0.100 ± 0.003 and 0.350 ±0.005,and the relative expressions of Caspase-3 protein were 0.020 ± 0.003,0.060 ± 0.003,0.020 ± 0.003 and 0.590 ±0.004,with significant differences among the 4 groups (F =136.681,2 717.391,44 471.429,P ＜0.05).The vitality of MiaPaca-2 cells of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =33.937,20.207,26.689,P ＜ 0.05).The relative expression of Caspase-8 protein of the TRAIL + Mcl-1 siRNA +group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =216.506,433.013,144.338,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =329.09,458.993,987.269,P ＜0.05).The vitalities of MiaPaca-2 cells of the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group,the TRAIL-Bcl-xL siRNA + group and the TRAIL+ Bcl-xL siRNA + group were 100.0％ ± 2.3％,87.2％ ± 4.1％,74.1 ± 3.7％ and 56.3％ ± 5.4％,and the relative expressions of Caspase-3 protein were 0.060 ±0.004,0.070 ± 0.003,0.060 ± 0.004 and 0.390 ± 0.003,with significant differences among the 4 groups (F =70.074,4 643.478,P ＜ 0.05).The vitality of MiaPaca-2 cells of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group and the TRAIL-Bcl-xL siRNA + group (t =24.416,41.170,18.136,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =285.788,554.256,190.526,P ＜ 0.05).Conclusion Triptolide could induce the apoptosis of MiaPaca-2 cells by inhibiting the expressions of Mcl-1 and Bcl-xL,sensitizing TRAIL and activating Caspase-8 and Caspase-3.

Objective To evaluate the long term results of coral bone used in anterior cervical interbody fusion. Methods The 180 patients (126 males and 54 females) who had undergone anterior interbody fusion in our hospital were followed up for 6 years. Their long-term imageological data, including changes in interbody height, interbody angle and range of movement, were analyzed and compared. Results In all the cases a solid fusion was achieved between coral reef and vertebra. The fusion was evaluated as successful in all the 180 patients. Conclusion The coral bone spacer provides the same solid fusion as autografting with iliac crest does.

One hundred and sixteen patients with lower limb fracture were screened by Doppler ultrasound for deep vein thrombosis(DVT) of bilateral lower extremities within the first 72 h, d7 and d21 after fracture. Results showed that DVT was detected in 31 (26. 7% ) out of 116 cases within 72 h; at d7 and d21 DVT was detected in 3 and 1 patient respectively with a cumulated DVT rate of 30. 2% in 3 weeks.Serial Doppler ultrasonography is of value in screening for DVT of the lower extremities in patients with lower limb fracture at early stage.

Objective To investigate the curative effect of mammotome minimally invasive operation in the treatment of benign breast disease and analyse its characteristics.Methods Two hundred and five cases of benign breast disease patients admitted to the hospital in time sequence were divided into the observation group (105 cases) and control group(100 cases).Observation group used mammotome minimally invasive rotary cutting operation,and the control group used conventional breast tumor resection.The surgical results of two groups of patients,tumor complete resection or not,whether the patients for surgery satisfaction were observed.Patients of two groups were compared with intraoperative and postoperative relevant indicators,and the postoperative complications.Results In the observation group,105 patients with mammotome spiral cutting knife surgery of breast masses were completely resected,the satisfaction of patients for surgery was 96.2％ (101/ 105),significantly higher than that of control group(81.0％ (81/100),x2 =4.187,P＜0.05).The operative time,blood loss,length of incision values of observation group were significantly lower than those in the control group((17.30±6.70) min vs (57.23 ± 8.96) min,(10.43 ± 5.14) ml vs (109.16 ± 13.45) ml,(10.27 ±0.06) cm vs (1.43±0.12) cm;t=18.741,26.167,11.421;P＜0.05),the postoperative recovery time and the scar length were significantly lower than the control group((4.1±2.5) d vs (8.0±3.5) d,(0.15±0.03) cm vs (1.21±0.46) cm;t =5.176,2.647;P＜0.05),breast deformation cases,tumor residual proporation were 1.9％(2/105) and 0(0/105),fewer than that in the control group(12.0％(12/100),9.0％(9/100);x2=6.721,11.470;P＜0.05).In control group breast deformation subcutaneous hematoma and ecchymosis,wound infection,skin damage,pain,pigment calm incidence were 2.9％ (3/105),5.7％ (6/105),1.9％ (2/105),2.9％(3/105),8.6％(9/105),2.9％(3/105),significantly lower than those in the observation group(9.0％(9/100),12.0(12/100),7.0％ (7/100),16.0％(16/100),23.0％(23/100),8.0％(8/100);x2 =2.164,3.071,2.467,6.194,6.177,2.642;P＜ 0.05).Conclusion Mammotome minimally invasive rotary cut scalpel compare with conventional breast tumor excision undoubtedly has a more significant curative effect and patient satisfaction and have the characteristics of operation safety,efficiency,beautiful,fewer complications,humanized.

Objective:In order to study mechanism of hematogenous metastasis of rectum cancer.Methods:8 specimens of human rectum cancer and 6 specimens of rectum in normal human were examined.The immunohistochemical SP method was employed in study of the expression of ICAM1,CEA and CD31 in the peritumoral rectum tissues and lymphy nodes.Results:The intercellular role in the adhesion molecule-1(ICAM-1)and carcinoma embryonic antigen(CEA) were expressed on the vascular endothelial cells of peritumoral rectum tissues and peritumoral lymph nodes in the rectum cancer.CD31 are expressed on the vascular endothelial cells of rectum tissues from normal human with the same intensity of cancer peritumoral rectum tissues.Conclusion:This study showed that ICAM-1 and CEA seemed to play a stable role in the adhesion effect between cancer cells and endothelial cells.It is not clear whether CD31 plays a role in the interaction between cancer cells and endothelial cells.

Objective To summarize the experience of surgical repair for Stanford type A aortic dissection after cardiac surgery.Methods From February 2009 to December 2011,11 patients who underwent previous cardiac surgery accepted the aortic surgery for Stanford type A aortic dissection.There were 8 males and 3 females.The range of age was from 29 to 64 years,the mean age was(52.27±9.90) years.In these patients,one patient had underwent ventrical septal defect,one patient atrial septal defect,nine patients aortic valve replacement.The interval between the two operations was 1-26 years.The types of aortic dissection was A1S(4 patients),A1C(1 patient),A2S(1 patient),A2C(4 patients),A3C(1 patient).All the patients underwent aortic surgery for aortic dissection.Results The time of cardiopulmonary bypass was 75-409 minutes,the mean value was(185.36± 99.67) minutes.Aortic cross clamp time was 37-203 minutes,the mean value was (84.09± 48.36) minutes.Total six patients needed deep hypothermia and selective cerebral perfusion time was 8-32 minutes.The mean value was(17.71 ± 9.48) minutes.One patient dead in the hospital and the mortality was 9％.The morbidity was 27％.Ten patients followed up 16-45 months.No aortic rupture,paraplegia and death were observed in follow-up time.Conclusion The delayed Stanford type A aortic dissection after cardiac surgery should be attached great importance and always need emergency surgery to save patients' life.The technique is demanding and risk is great for surgeons and patients.For the patients who suffered aortic valve disease combined with dilation of ascending aorta larger than 4.5 cm,the ascending aorta also should be repaired while aortic valve replacement is performed,which could avoid delayed aortic dissection in the future.

Objective Acute type A aortic dissection associated pregnancy severely threatens the lives of both the mother and her ferus.We retrospectively reviewed our clinical experience with this life-threatening condition in six cases.Methods Between January 2007 and February 2012,6 women with acute type A aotic dissection associated pregnancy were treated by our group,with an average of 3 1 years (range 24 -37 weeks)and a mean gestation weeks of 24.5 (range,12 -38 weeks ).The etiology was Marfan syndrome in 4 cases and gestational hypertension in 2.The pathology was the modified Stanford type A3S in I case,A2C in 2 and A3C in 3.- Five patients were treated surgically and 1 medically.Surgical operations were performed under hypothermic cardiopulmonary bypass or deep hypothermic circulatory arrest,including Bentall procedure in 1case,Bentall + Sun's procedure in 2,ascending aortic replacement + Sun's procedure in 2.Results The woman treated medically and her fetus died from aortic rupture 9 days after admission.The cardiopulmonary bypass and cross clamp time and circulatory arrest time averaged 167 rninites(range,75 -210 minites) and 98 minites(range,83 - 145 minites) and 23.5minites(range,19 -27 minutes),respectively.Five patients treaed surgically survived the operation.Three fetuses survived rand two fetuses died.After a mean follow-up of 2.2 years (range,1 - 3.5 years ),5 patients were doing well.CT angiogram detected nonmal aortic and valvular structures,with no signs of distal dilation.Three babies were normal in development and neurocognitive functios.Conclusion Palients with aortic dissection associated with pregnancy should be operated on ugently and medical treatment carries high risks of aortic rupture and maternal and fetal death.Methods of surgical repair,peffusion techniques and delivery should be chosen based on the underlying aortic pathology and gestational age,so as to maximize the safety of the mother and her baby.

Objective Virus-specific RNA interference (RNAi) is a powerful inhibitor of gene expression and replication of HBV. It is known to have high efficiency, specificity, and few side effects. We wanted to evaluate the effects of siRNA silencing HBV replication on the growth of hepatocellular carcinomatic(HCC) cells to find out an ideal method for treatment of HCC. Methods We transfected siRNA into HepG2.2. 15 cells (HCC cell inserting HBV gene) and detected the HBsAg and HBV DNA copies for evaluating the inhibitory effects of siRNA. Then we evaluated cell growth and self-renewal ability after transfection of siRNA by MTT. Results The HBsAg level and HBV DNA copies were reduced after the transfection of siRNA, the highest inhibition rate was 83.9%,while the inhibition rate of HBV DNA copies reached 73. 4%. The siRNA group's growth ability and self-renewal rate were lower than the control group in 5 days. Conclusion siRNA can effectively inhibit HBV replication and expression in HepG2.2.15 cells and silencing HBV replication can inhibit HepG2.2.15 cell's growth and self-renewal.

Objective To investigate the protectve effects and underlying mechanisms of deferroxamine on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal rat were used in a model of glutamate induced neurotoxicity.There were two experimental groups.Neurons were pretreated with deferroxamine before glutamate in the deferroxamine group, and neurons were treated with glutamate only in the control group.The morphological change was examined under microscope.Hoechst 33342 DNA staining method was used to study the ratio of condensed nuclei.The levels of lactate dehydrogenase (LDH), malonaldehyde (MDA) and hydroxyl radical were determined using biochemistry.The change in calcium signal was detected using microfluorescent technique.Results The neurons pretreated by deferroxamine had intact morphology with the ratio of condensed nuclei at 14% ± 6% compared to 58% ± 6% (t= 8.98, P ＜0.01 ) in the control group.LDH level was (36.42 ± 8.99) U/L in the deferroxamine group and was (68.06 ± 11.26) U/L in the control group ( t =3.25,P＜0.05).The respective levels of hydroxyl radical were (34.21 ±4.23) U/L and (47.06 ±8.79) U/L (t = 3.11, P ＜0.05 ).The respective levels of MDA were (12.26 ± 2.78 ) nmol/mg and (28.86±5.19) nmol/mg(t =4.88,P＜0.01).Conclusion Deferroxamine can protect neurons from glutamate induced damage.The mechanisms include an inhibition of Ca2+ overload and reduction in the levels of MDA and hydroxyl radicals.

AIM: To investigate the effects of heptanol preconditioning on the changes of structure, function and connexin 43 (Cx43) content in mitochondria in a rabbit model of myocardial isehemia-reperfusion (IR) injury. METHODS: In anesthetized open-chest rabbits, the left anterior descending artery (LAD) was occluded for 30 min and reperfused for 4 h. Sixty-four rabbits were randomly divided into 4 groups (n=16 in each group): sham operation group (sham group), ischemia-reperfusion group (IR group), ischemic preconditioning group (IP group) and heptanol preconditioning group (HT group). All rabbits in the 4 groups were killed 4 h after reperfusion. Myocardial infarct size was determined at the end of the experiment. Mitochondria was isolated by centrifugations. The ultrastructural changes of the mitochondria were observed under electronic microscope. The mitochondrial membrane potential, Ca~(2+) concentration, MDA content and SOD activity of myocardial mitochondria were also examined. The content of mitochondria Cx43 was detected by Western blotting. RESULTS: Compared to IR group, the myocardial infarct size was significantly reduced in IP (18.97%±2.80%) and HT (19.97%±3.80%) groups, the damage of mitoehondrial ultrastructure was milder (P<0.05), mitochondrial membrane potential was significantly higher and Ca~(2+) concentration was much lower (P<0.01) in IP group and HT group. No significant difference of MDA content and SOD activity in myocardial mitochondria between IR group and HT group was found. However, MDA content were much lower and SOD activity was significantly higher in IP group as compared to IR group (P<0.01). Compared to sham group, the mitochondria Cx43 expression was distinctly decreased compared to IR group (P<0.05) and no significant difference was found between IP group and HT group (P>0.05). CONCLUSION: Heptanol preconditioning protects myocardium from ischemia-reperfusion injury. The mechanism may be related to increasing in mitochondrial membrane potential, alleviating Ca~(2+) overload in myocardial mitochondria and attenuating the decrease in mitochondria Cx43 expression induced by isehemia-reperfusion.