Objective:To investigate whether PUMA gene transfection can increase sensitivity of pancreatic cancer cells (PC) to 5-FU-induced apoptosis. Methods: PUMA-pCEP4 containing full length PUMA cDNA or pCEP4 was transfected into human pancreatic cancer cell line AsPC-1 by lipofectamine transfection, G418 selection was used to select positive cells. AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were separately treated with serial concentrations of 5-FU(0.01-100 ?mol/L). MTT assay was used to determine the cell survival rate in each group and IC50 of 5-FU was calculated. TUNEL,FCM and DNA ladder observation were employed to study cell apoptosis. Western blotting was performed to detect the expression of PUMA protein. Results: The 5-FU IC50 values of AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were (12?1.9)?mol/L,(1.6?0.4)?mol/L and (10.4?1.6) ?mol/L, respectively, with the sensitivity of AsPC-1/PUMA cells increased by 7.5 folds. 5-FU induced cell apoptosis of AsPC-1 cells in a dose-dependent manner, with the apoptosis of AsPC-1/PUMA cells more prominent than those of AsPC-1 and AsPC-1/pCEP4 cells. Low concentration of 5-FU (0.1 ?mol/L) induced few apoptosis of AsPC-1/pCEP4 cells([1.14?0.28]%) and AsPC-1 cells ([0.9?0.23]%), and induced apoptosis in AsPC-1/PUMA cells([6.47?1.42]%). High concentration of 5-FU (1.0 ?mol/L) induced apoptosis in all groups, with that in AsPC-1/PUMA cells([34.54?9.36]%) significantly higher than those in AsPC-1/pCEP4 cells([15.8?5.15]%) and AsPC-1 cells ([12.8?3.74]%, both P

Background and purpose:Members of the NF-kappaB family of transcription factors could activate bcl-2 anti-apoptotic genes at the transcriptional level and then regulate cell apoptosis.The p53 tumor suppressor gene has a clear role!in both the regulation of cell cycle and expression of apoptosis-related bcl-2 family.There were few reports about the expression of NF-?Bp65,bcl-2,bcl-xL and their relationships with p53 and cell apoptosis in pancreatic cancer.The present study was designed to analyze the expression of the antiapoptotic molecules nuclear factor kappaB(NF-?Bp65),bcl-2 and bcl-xL in pancreatic ductal carcinoma(PC) and their correlations to the apoptosis index and p53 expression.Methods:Immunohistochemical analyses of P53 protein was performed on 25 cases of pancreatic ductal carcinoma and 9 cases of normal pancreas;Western blot was used to evaluate NF-?Bp65 protein and RT-PCR for bcl-2 mRNA and bcl-xL mRNA of 25 cases of pancreatic ductal carcinoma and 9 NP cases.The apoptosis was determined by TUNEL.Results:The positive rate of P53 protein was 56%(14/25)in PC tissues,significantly higher than that in NP tissues(P

Objective To study the effect of PUMA gene mediated by recombinant adenovirus vector combined with radiation on the pancreatic carcinoma. Methods The PANC-1 cells were infected with Ad-PUMA (MOI = 10, 50 and 100, respectively) for 48 h. The expression of PUMA mRNA and protein was detected by RT-PCR and Western blot, respectively. PANC-1 cells were divided into 4 groups: control group, transfection group, irradiation group and combined treatment group. The cell growth inhibition rate and apoptotic rate of PANC-1 cells were assessed by MTT assay and flow cytometry. Human pancreatic carcinomas were transplanted subcutaneously in nude mice, which were randomized into 4 groups: control group, transfection group, irradiation group and combined treatment group. Tumor growth rate and apoptotie index at different time points were recorded in 35 days. Results The expression of PUMA mRNA and protein was increased with the increase of MOI of Ad-PUMA, which was does-dependant (MOI = 10, mRNA = 0.46 ± 0.02, protein = 0. 75 ±0.09;MOI=50, mRNA= 1.12±0.09, protein = 1.01 ±0.18;MOI= 100, mRNA= 1.50±0.08, protein=1.80 ± 0.15 ;P < 0.05). The proliferation of PANC-1 cells was suppressed significantly when transfected by Ad-PUMA in a dose-dependent manner(r = -0.986 55), which was more significant combined with radiation (r = -0.971 26, P < 0.05). Meanwhile, the apoptotie rate was increased in the same manner [for pre- and post-irradiation,which was (45.4 ± 5.26) % and (73.2 ± 6.62) %, respectively, P < 0.05]. From 7 to 35 d after PUMA gene transfection and radiotherapy, the tumor growth was significantly slower than those of irradiation group, transfection group and control group [35 d after therapy, the volume of tumor was (19.82 ± 6.45)mm3 ,(39.5 ± 9.23)mm3 , (33.6 ±3 10.3)mm3 and (52.0 ± 11.43)mm3 , respectively, P < 0.05]. And the apoptotic index was increased in the same manner (AI = 0.43 ± 0.05, 0.29 ± 0.10, 0.24 ± 0.05 and 0.00 ± 0.00, respectively, P < 0.05). Conclusions Recombinant adenoviral-mediated PUMA gene combined with irradiation could increase the cell-killing effect on pancreatic carcinoma. It is better than that of either one kind of therapy.

Objective To explore the effect of salvianolate combined with Qumei trimetazidine on cardiac function in patients with chronic heart failure.Methods Seventy-four patients with chronic heart failure were randomly divided into treatment group and control group (37 cases per group).Patients in control group were treated with the regular treatment scheme including digitalis,diuretics,vasodilators,angiotensin converting enzyme inhibitor(ACEI),angiotensin receptor blockers (ARB) or β blocker therapy for 24 weeks treatment.Patients in treatment group were given the regular treatment scheme plus salvianolic acid and Qumei trimetazidine treatment,of which,the dose of salvianolic was 0.2 g into 5％ glucose injection 250 ml or 0.9％ sodium chloride injection 250 ml by intravenous injection,1 times/day,and Qumei trimetazidine for 20 mg,3 times/day,for 24 weeks.Cardiac function was observed in patients of two groups before and after treatment.The level of brain natriuretic peptide (BNP) was measured.Results Heart function were improved,the total effective rate in treatment group was 91.9％ (34/37),higher than that of control group (70.3％ (26/37),x2 =5.638,P ＜ 0.05).In treatment group,left ventricular ejection fraction (LVEF),stroke volume (SV),cardiac output (CO) of patients after treatment were (52 ± 7) ％,(65.10 ± 12.87) ml,(5.65 ± 1.18) L/min respectively,significant different from that before treatment ((39 ±5)％,(46.53 ± 12.14) ml,(4.79 ± 1.02) L/min,and the differences were statistic significant (t =9.192,6.384,3.352,P ＜ 0.05).Meanwhile,in treatment group,systolic pressure,diastolic pressure,heart rate,left ventricular end diastolic diameter (Dd),left ventricular diastolic posterior wall thickness(PWT),interventricular septal thickness (IVST),left ventricular mass (LVMW),plasma brain natriuretic peptide of patients after treatment were (105 ± 8) mmHg,(75 ± 9) mmHg,(76±8) time/min,(48.7 ±3.7) mm,(9.1 ±1.4) mm,(8.7 ±1.2) mm,(170±59) g,(104.1 ±19.5) ng/L respectively,significant different from that of before treatment((134 ± 12) mmHg,(84 ±8) mmHg,(118 ±11) time/min,(55.2 ±7.8) mm,(11.7 ±2.3) mm,(10.5 ±2.4) mm,(228 ± 111) g,(568.7±179.5) ng/L t=-12.231,-4.546,-18.782,-4.579,-5.874,-4.080,-2.806,15.652,P ＜ 0.01).The same trend was seen in control group in terms of LVEF,SV,systolic blood pressure,heart rate,PWT,plasma BNP before and after treatment(LVEF:(38 ±6)％ vs.(43 ± 8)％ ;:(46.76 ± 11.80) ml vs.(58.69 ± 11.58) ml; systolic blood pressure:(132 ± 10) mmHg vs.(116 ± 11) mmHg; heart rate:(116 ± 10) time/min vs.(77 ±9) time/min;PWT:(11.5 ±2.6) mm vs.(10.4 ±2.0) mm;plasma BNP:(570.2 ± 177.3) ng/L vs.(211.6 ± 21.2) ng/L;t =3.041,4.389;-6.546,-17.632,-2.039,12.21 ;P ＜ 0.05 or P ＜ 0.01).Moreover,after treatment,systolic pressure,diastolic pressure,LVEF,SV,CO,Dd,PWT,IVST,LVMW,plasma brain natriureticpeptide in treatment group were significantly better than that of control grouo (t =-4.919,-2.867,5.510,2.252,2.581,-2.319,-3.238,-3.628,-2.231,-22.701,P ＜0.01 or P ＜ 0.05).Conclusion The effect of salvianolate combined Qumei trimetazidine on treating chronic heart failure is significant,and there is a reverse effect on the left ventricle.

Objective To investigate the effect of salvianolate on chronic heart failure in patients with cardiac function and plasma brain natriuretic peptide effect.Methods Sixty-eight cases with chronic heart failure patients were randomly divided into treatment group and control group (34 cases for each group).Patients in control group were given the conventional treatment,in treatment groups were given conventional treatment plan plus salvianolic acid at dose of 0.2 g added 5％ glucose injection 250 ml (or 0.9％ sodium chloride injection 250 ml),1 times a day for 12 weeks.The cardiac function was recorded and brain natriuretic peptide level was measured before and after treatment.Results After 12 weeks of treatment,the total efficiency in treatment group was 91.2％ (31/34)) higher than that in control group(70.6％ (24/34)),and the difference was statistically significant (x2 =9.399,P ＜ 0.01).Before treatment,the left ventricular ejection fraction (LVEF),stroke volume(SV),cardiac output(CO) in treatment group were (38 ±6)％,(44.64 ± 11.03) ml,(4.81 ± 1.03) L/min respectively,differed from that after treatment ((51 ± 8) ％,(63.21 ± 11.94) ml,(5.67 ± 1.17) L/min),and there were significant differences between before and after treatment (t =-7.580,-8.975,-3.233 respectively; P ＜ 0.01).The levels of systolic blood pressure,diastolic blood pressure,heart rate,left ventricular end-diastolic internal diameter (Dd),the left ventricular diastolic wall thickness (PWT),diastolic interventricular septal thickness (IVST),left ventricular mass (LVMW),brain natriuretic peptide in treatment group before treatment were (131 ± 11) mmHg,(85 ± 7) mmHg,(116 ± 9) times/min,(55.1 ± 7.9) mm,(11.8 ± 2.4) mm,(11.4 ± 2.3) mm,(231 ± 112) g,(572.9 ± 183.6) ng/L respectively,significant differed from those of after treatment((104 ± 7) nmHg,(76 ± 8) mmHg,(75 ± 7) times/min,(48.8 ± 3.9) mm,(9.2±1.3) mm,(8.9± 1.1) mm) (172 ±57) g,(101.8 ± 18.5) ng/L respectively),and the differences were significant (t =12.075,4.937,20.961,4.169,5.556,5.721,2.738,14.886 ; P ＜ 0.01).The levels of LVEF,SV in control group before treatment were (37 ±7)％ and (44.87 ± 10.82) ml,differed from those of after treatment((42 ± 9)％ and (56.70 ± 10.60) ml;t =-2.556,-4.554;P ＜ 0.01).The systolic blood pressure,heart rate,Dd,IVST,plasma brain natriuretic peptide in control group before treatment were (130 ±12) mmHg,(114 ± 10) times/min,(54.8 ± 8.7) rmm,(11.3 ± 2.6) mm,(574.1 ± 181.4) ng/L respectively,significantly differed from those of after treatment ((115 ± 9) mmHg,(76 ± 8) times/min,(50.6 ±8.3) mm)(9.9±1.3) mm,(215.7 ±23.2) ng/L;t=5.830,17.304,2.037,2.806,11.427;P＜0.01 or P ＜ 0.05).The levels of systolic blood pressure,diastolic blood pressure,LVEF,SV,CO,PWT,IVST,plasma brain natriuretic peptide in treatment group were better than that in control group (t =-4.601,-3.093,4.358,3.253,2.802,-3.066,-3.425,-27.985,P＜0.01).Conclusion Salvianolate is proved to be better drug on treating chronic heart failure curative with left ventricular reverse effect and less adverse reaction.

Objective To assess the clinical efficacy of minimally invasive internal fixation combined with percutaneous kyphoplasty (PKP) in treatment of thoracolumbar burst fractures in the elderly.Methods Twenty-one cases of neurologically intact thoracolumbar burst fractures treated by PKP between January 2007 and December 2008 were included in this study.There were 8 males and 13 females at age of 65-78 years (means,70.6 years).Mean period from injury to operation was (3.7 ± 1.1) days (range,3-7 days).The injured segments included Ti1 in two cases,T12 in six,L1 in eight and L2 in five.Kphosis Cobb' s angle,correction degree of kyphosis angle,correction loss of kyphosis,perioperative indicators,visual analogue scale (VAS) and Oswestry disability index (ODI) were evaluated after operation.Results All the cases were followed up for a period of 26-56 months (mean 34.2 months).Operation averaged 98.7 minutes (range,80-120 minutes) and showed mean blood loss of 32.8 ml (range,30-85 ml).Ambulation started at mean 18.2 hours after operation (range,8-19 hours).VAS averaged (0.9 ± 0.6) points at postoperative one week.Postoperative X-ray films revealed mean 10.4° correction of kyphotic Cobb angle,followed by mean 1.8 °of loss in the follow-up longer than 24 months.According to hyperextension and hyperflexion radiographies,there was no abnormal activity of fixed segments,implant loosening and breakage or adjacent vertebral fractures.Conclusion Minimally invasive internal fixation combined with PKP can relieve pain and restore function in a short time and is thought to be a preferable treatment option for thoracolumbar burst fractures in the elderly.

Objective To study a case series of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns to analyze the causes of its occult symptoms and explore the injury mechanism,imaging diagnosis and operative reduction of the fractures.Methods A retrospective study was carried out on data of 39 patients with thoracolumbar flexion-distraction fractures admitted into our hospital from May 2003 to December 2006.Of all,there were 17 patients with compression of anterior and central columns.The diagnostic value of imaging examinations such as X-ray,CT and MRI was evaluated.All patients were treated with posterior operation,in which simple distraction of the anterior and central columns was followed by compression and reduction of the posterior column from the rear axial direction.The clinical efficacy and safety of operation were assessed.Results The patients with compression of anterior and central columns accounted for 43.6% of all thoracolumbar flexion-distraction fractures.Before operation,six patients were misdiagnosed as simple compression fracture.Various kinds of imaging examinations detected the bone injury and/or ligaments complex injury of the posterior column at different degrees.The positive results on X-ray,CT,spiral CT multi-planar reconstruction (MPR)and MRI were in 8,7,11 and 17 patients respectively.After simple posterior distraction in 17 patients,there occurred over distraction in eight patients who were cured with posterior recompression plus reduction.Conclusions The posterior column injury of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns is relatively occult and easy to be misdiagnosed as simple compression fracture,when MPR CT and MRI are helpful for diagnosis.The operation with twostep reduction of axial distraction followed by compression can attain satisfactory and safe results.

Objective To investigate the effect of ischemic preconditioning (IPC) on expression of intracellular adhesion molecule-1 (ICAM-1) in brain tissues following traumatic brain injury (TBI) in rats.Methods Sixty male SD rats weighing 220-250 g were randomly divided into three groups (n =20 for each):sham operation group,TBI group,and IPC group.Cerebral IPC models were induced by transient occlusion of the bilateral common carotid arteries; TBI models were induced by Feeney's freefalling method; rats in sham operation group were only performed exposure of dura of the right parietal lobe.Ten rats were sacrificed respectively at 6 and 72 hours after TBI and injured brain tissues were harvested to estimate wet/dry weight (W/D) ratio for the brain,determine ICAM-1 expression by immunohistochemistry and perform microscopic examination.Results Brain W/D ratio was significantly increased in TBI group compared with sham operation group (6 h:4.2 ± 0.4 vs 2.7 ± 0.4 ; 72 h:5.0 ±0.1 vs 3.1 ± 0.2,P ＜ 0.05).ICAM-1 expression was up-regulated in TBI group compared with sham operationgroup (6h:25.4±3.5vs8.6±1.3; 72 h:36.5±5.4 vs8.4±1.6,P＜0.05).W/D ratio was significantly decreased in IPC group compared with TBI group (6 h:3.5 ±0.6 vs 4.2 ±0.4; 72 h:3.7 ± 0.4 vs 5.0 ± 0.1,P ＜ 0.05).ICAM-1 expression was down-regulated in IPC group compared with TBI group (6 h:16.5 ± 2.7 vs 25.4 ± 3.5 ; 72 h:24.3 ± 4.6 vs 36.5 ± 5.4,P ＜ 0.05).Milder injury to brain tissues was observed in IPC group than in TBI group.Conclusion IPC can attenuate TBI in rats by down-regulating the expression of ICAM-1.

Objective To compare the Roussouly classification of old thoracolumbar fractures combined with post-traumatic kyphosis with that of normal adults, evaluate the effect of different kyphosis angles on sagittal curvature of the thoracolumbar spine so as to provide a theoretical basis for the clinical status and clinical treatment plan of old thoracolumbar fractures combined with post-traumatic kyphosis.Methods A retrospective case control study was made on 49 cases of old thoracolumbar fractures with post-traumatic kyphosis treated from January 2014 to December 2015 (fracture group).Damaged segments of the spine were T11 in four cases, T12 in 14, L1 in 25 and L2 in six.Another 52 normal adult volunteers were chosen as controls.Whole spine X-ray film of the two groups was taken, and the difference in Roussouly classification of the sagittal spine curvature was compared between the two groups.Patients in fracture group were separated into subgroups depending on the Cobb angle (0°-10°, 10°-20°, 20°-30°, 30°-40°, respectively), and the difference in Roussouly classification was compared among subgroups.Results Roussouly classification in control group was five cases of type 1, 12 tpye 2, 25 type 3 and 10 type 4.Results in fracture group was 14 cases of type 1, 11 type 2, eight type 3 and 16 type 4.There were significant differences in Roussouly types between the two groups (P<0.05).According to Roussouly classification, there were two cases of types 1 and 2 and seven cases of types 3 and 4 in subgroup of Cobb angle between 0°-10°;four cases of types 1 and 2 and 10 cases of types 3 and 4 in subgroup of Cobb angle between 10°-20°;12 cases of types 1 and 2 and four cases of types 3 and 4 in subgroup of Cobb angle between 20°-30°;seven cases of types 1 and 2 and three cases of types 3 and 4 in subgroup of Cobb angle between 30°-40°.There were significant differences in Roussouly types among subgroups (P<0.05).Further, Roussouly types 1 and 2 were significantly increased when the Cobb angle was >20° (P<0.05).Conclusions Post-traumatic kyphosis in old thoracolumbar fractures affects spine sagittal curvature.Patients with sagittal spinal imbalance (Roussouly types 1 and 2) are more with increased Cobb angle.Moreover, Cobb angle >20°may be the reference basis for clinical intervention.

0.1). Plasma PAF-AH activity in the patients with psoriasis was significantly lower than that in the healthy controls(P