Objective To study the relationship of expression of nuclear factor kappab gene and MMP-2mRNA and with lymph node metastasis of pancreatic cancer.Methods The expression of NF-?B was investigated by Western blot in normal pancreatic(NP) samples of 9 cases and pancreatic cancer(PC) samples of 45 cases(lymph node metastasis 30 cases,non-lymph node metastasis 15 cases).The expression of MMP-2mRNA was detected through semiquantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results Of 45 PC cases,the positive expression rate of p65 and MMP-2mRNA was 66.7%(30/45) and(57.7%)(26/45),respectively.Of 30 cases of lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA was 83.3%(25/30)and 73.3 %(22/30),respectively.Of 15 cases of non-lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA is 33.3%(5/15) and 26.7%(4/15),respectively,and the expression of p65 gene was positively correlated with MMP-2mRNA(expression)(r=0.743,P

objective To construct the recombinant adenovirus containing p53 up-regulated modulator of apoptosis(Ad-PUMA)and investigate its growth inhibition effect on pancreatic callCer cells in vitro and in vivo.Methodls Ad-Easy system was used to construct Ad-PUMA by recombination in E.coli.The virus was Dackaged in 293 cells and subsequently identified valid.The AsPC-1 cells were infected with AdPUMA.Before and after Ad-PUMA infection,the expression of PUMA protein wag investigated by western blot,the inhibition rate of AsPC-1 cells was examined by MTY assay.The in vivo tumor suppressive effect was detected in nude mice with human AsPC-1 xenograft.PUMA protein and the apoptosis of AsPC-1 xenograft were detected by western blot and TUNEL(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling)method.Results In vitro,the expression of PUMA protein was increased with titer of Ad-PUMA,the proliferation of AsPC.1 cells were suppressed,significantly,and the effect was in a viral dose-dependent nlanner.In vivo,the growth in nude mice of AsPC-1 infected with Ad-PUMA was significantly inhibited with an inhibition rate of 44.2%.The expression of PUMA was significantly up-regulated,and the apoptosis index wa8 significantiv increased in tumor after Ad-PUMA infection as determined by western blot and TUNEL.Conclusion The expression of PUMA call inhibit the proliferation of pancreatic cancer in vitro and in vivo,and may be used 88 a potential tool for cancer therapy

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 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

Objective To study surgical techniques and clinical effects of minimally invasive excision of high rectal carcinoid tumors with anus preservation. Methods We conducted transanal local excision of rectal carcinoid tumors in 6 cases by using self-made proctoscope and laparoscopic instruments from August 2002 to January 2005. The tumors were located on the depth of 9~12 cm from the anal verge, and excision margin was 0.5~1 cm from the tumors. Results All the operations were performed successfully. A follow-up for 3~28 months (mean, 15 months) found no recurrence, metastasis, rectal stenosis, or other short- or long-term complications. The postoperative hospital stay was 3~6 days. No analgesics were required. Conclusions Transanal local excision of high rectal carcinoid tumors under proctoscope is safe, reliable, minimally invasive, and cost-effective.

Objective:To investigate the effect of P53 up-regulate modulator of apoptosis(PUMA) on the apoptosis of pancreatic carcinoma BxPC-3 cells and the possible mechanism.Methods: BxPC-3 cells were infected with recombinant adenovirus containing PUMA gene(Ad-PUMA) at 100 MOI for 0-96 h.Apoptosis of BxPC-3 cells was examined by FCM.Expressions of PUMA,Bcl-2,Bax,Cytochrome C and Caspase-3 proteins in BxPC-3 cells were detected by Western blotting.Bax expression in the cytoplasm and mitochondrion and Bax oligomer expression expression in BxPC-3 cells were determined by Western blotting.Results: Apoptosis rates of BxPC-3 cells were significantly increased with the time of Ad-PUMA infection,and peaked after 48 h.Ad-PUMA infection increased the expressions of PUMA,Cytochrome C and Caspase-3 proteins in BxPC-3 cells,and decreased the expression of Bcl-2 protein.Apoptosis rate of BxPC-3 cells after Ad-PUMA infection was correlated with PUMA expression.Ad-PUMA did not affect the expression of total Bax protein in BxPC-3 cells,but Bax expression in cytoplasm was dramatically decreased after infection,and Bax expression in mitochondrion was markedly increased.Furthermore,Ad-PUMA infection induced Bax oligomerization in BxPC-3 cells.Conclusion: PUMA can promote apoptosis of pancreatic carcinoma cells through mitochondrion pathway.

Objective To investigate the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling pathway in the endometrium of women with polycystic ovary syndrome (PCOS) and its effect and significance in the cause of hyperplasia and carcinoma;and investigate the factors which affect the activation of the MAPK/ERK signaling pathway. Methods Collected 52 patients diagnosed as PCOS who were taken dilation and curettage of uterus as study, while 32 non-PCOS patients matched as control group. Serum hormonal parameters, fasting blood glucose and insulin were measured in all patients. The PCOS patients were sub-group as insulin resistance group and non-insulin resistance group; all the patients were carried out pathology inspection of endometria, and the PCOS patients were sub-group as endometrial hyperplasia and carcinoma group and normal endometrium group based on the outcome of pathology inspection. Western blot were performed to detect the expressions of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2), the activation of ERK1/2. Results (1)The expression of pERK1/2 [(61 ±13)%] in the endometrium in PCOS group was higher than that in the control [(44 ±10)%, P ＜0.01]. (2)The expression of p-ERK1/2 was significantly increased in group of endometrial hyperplasia and carcinoma [ ( 70 ± 11 )% ] compared to that in group of normal endometrium [ (55 ± 10)% ,P ＜ 0.01 ], while there were significant difference between group of insulin resistance [ (63 ± 13 )% ] and group of non-insulin resistance [ (55 ±7)%, P ＜0.01 ]. (3) Fasting insulin level, insulin area under the curve and body mass index were related to the expression of p-ERK1/2 in endometrium with PCOS, the correlation coefficient were 0.447, 0.456 and 0.381, respectively ( all P ＜ 0.01 ). Conclusions The MAPK/ERK signaling pathway in endometrium with PCOS was overactivation, which was related to the endometrial hyperplasia and carcinoma; while the activation of MAPK/ERK signaling pathway were effected by insulin resistance and hyperinsulinemia.

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 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 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.