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 To investigate the effects of ischemic postconditioning on apoptosis, structural and functional changes of mitochondria induced by myocardial isehemia/reperfusion (I/R) injury of rabbits and potential mechanism. Methods Eighty healthy rabbits were divided randomly into five groups: sham operation group ( Group Sham) , ischemic reperfusion group (Group IR) , ischemic preconditioning group (Group IP) , ischemic postconditioning group (Group PC) and 5-HD plus ischemic postconditioning group (Group PC +5-HD). All rabbits in the five groups were killed 4 h after reperfusion. The hearts were quickly collected for microscopy by TUNEL. We observed ultrastructural changes of myocardium under electron microscope and examined mitochondrial membrane potential and Ca~(2+) concentration, MDA content and SOD activity of myocardial mitochondria. Results Compared with group IR, the damage of mitoehondrial ultrastrueture was milder, the apoptosis rate decreased and Ca concentration and MDA content were much lower in group IP and group PC ( P < 0. 05 ). Mitochondrial membrane potential and SOD activity of myocardial mitochondria in group IP and group PC was significantly higher than that in group IR(P<0.05). The protective effect of PC against I/R injury was partially counteracted by 5-HD .Conclusion Ischemic posteonditioning can protect the heart from I/R injury, this is supported by improvement mitochondrial ultrastructure and by decreasing apoptosis, increasing mitochondrial membrane potential and SOD activity, alleviating Ca~(2+) overload and decreasing MDA content in myocardial mitochondria. The cardio protective effects may be explained by mitochondrial ATP sensitive potassium channel.

Objective To make comparisons of the three models of acute and chronic rheumatic carditis to find out an optimal animal model.Methods AntigenⅠwas a emulsifier mixed by complete freund’ s adjuvant( CFA) and Group A streptococcus(GAS).AntigenⅡwas mixed by incomplete freund’s adjuvant(IFA) and GAS.Female Lewis rats were randomly divided into four groups: A, B, C treatmeat groups were immuned with antigenⅠat the foot pad firstly. Subsequently, rats in group A、B、C were injected antigenⅠ, antigenⅡand activated GAS respectively to make the models of RHD.Rats in control group D were immunized with the same protocol outlined as treatment groups but without GAS. Respectively 7, 12, 24 weeks the rats were sacrificed 24 ( each group was 6).The blood biochemical item and Hematoxylin-eosin( HE) staining of hearts were detected.Results In group C the mortality was 25%.In group A, the incidence of carditis was the highest.Histopathological manifestations of group A, C was not only revealed acute damage such as inflammatory cell infiltrate as well as group B, but also the Aschofflike cells in the myocardial cells interstitial.But in group A and C there had a great degree of the inflammatory cells infiltration than group B.At 24th week rats in group A detected the rate and degree of valve fibrosis in chronic damage were higher than group B and C.None of rats in group D presented carditis or valvulitis.Conclusion In group A, giving the GAS with continuous stimulation after using the mixed emulsification of CFA and GAS to immune Lewis rats for five times was a appropriate method which could provide an optimal animal model for experimental study of acute and chronic rheumatic heart disease.

The cyclin-dependent protein kinase 9 (CDK9) is a member of the family of cyclin-dependent protein kinases.Different from other CDKs,CDK9 mainly works on the transcription regulation and has no affects on the cell cycle progress.There are many kinds of CDK9 inhibitors in the clinical research.The detailed structure and action mechanism of CDK9,its difference of protein structure from other CDKs,several selective or nonselective CDK9 inhibitors,as well as their structure-activity relationship (SAR) are discussed in this paper.

Objective To assess the left ventricular systolic function after renal sympathetic denervation(RSD) in beagles with heart failure by layer-specific two-dimensional speckle tracking imaging.Methods Twenty-three beagles were randomly divided into control group (8 dogs),URSD group (8 dogs)and RSD group (7 dogs).Control group was implanted with pacemakers without pacing,URSD group was implanted with pacemakers and underwent 3 weeks of rapid right ventricular pacing,and RSD group underwent catheter-based RSD bilaterally and was simultaneously implanted with pacemakers.All standard measurements were obtained from the left ventricular short-axis views at the levels of mitral annulus,papillary muscle and apex,and apical four-chamber,two-chamber,and long-axis views.The peak systolic longitudinal strain and circumferential strain of each layer of the left ventricle were measured by layerspecific two-dimensional speckle tracking imaging.Results After 3 weeks,all of the dogs in both the URSD and RSD groups showed lower left ventricular diastolic and systolic function compared with those in the control group,however,the dogs in the RSD group had a higher LV ejection fraction (LVEF) than the dogs in the URSD group(P ＜ 0.05).Compared with the control group,the global longitudinal strain,circumferential strain and radial strain all decreased significantly in URSD and RSD groups.The longitudinal strain of endo-and mid-cardium and the circumferential strain in endo-cardium were significantly higher in RSD group than those in URSD group(P ＜0.05).Conclusions Myocardial systolicfunction is impaired in dogs with heart failure,layer-specific two-dimensional speckle tracking imaging can provide effective tools to assess the left ventricular systolic function.Renal sympathetic denervation can improve the myocardial systolic function in dogs with heart failure.

Objectives To assess the effect of nicorandil and trimetazidine on myocardial microcirculation reperfusion in patients with NSTEMI after elective PCI. Methods 80 patients with NSTEMI were randomly assigned to four groups: normal medicine group (CON), nicorandil group (NIC), trimetazidine group (TMZ) and Combination group (NIC+TMZ). The coronary angiography and PCI were performed after 10 days. MCE was taken since 72 hours after operation. Results There were no significant differences in baseline characteristics between the four groups (P > 0.05). The A, β and A ·β of group NIC were significantly higher than group CON (P < 0.05). There were no significant differences between the group TMZ and the group CON (P>0.05). The A,βand A·βof group NIC+TMZ were significantly higher than group NIC or TMZ (P<0.05). Conclusions Nicorandil can improve the situation of myocardial reperfusion after elective PCI in patients with NSTEMI, and it has some synergetic effect when combined with trimetazidine.

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.