不合并消化道梗阻的晚期胰头癌441例临床诊治分析

目的 分析不合并消化道梗阻的晚期胰头癌的临床资料,探讨治疗策略.方法 回顾性分析2001年1月至2010年12月收治的441例不伴消化道梗阻的晚期胰头癌患者的临床资料.结果 所有患者均行手术治疗,其中行胆囊-空肠Roux-en-Y吻合术(A组)101例,胆总管-空肠Rouxen-Y吻合术(B组)133例,胆囊-空肠Roux-en-Y吻合+胃-空肠吻合术(C组)83例,胆总管-空肠Roux-en-Y吻合+胃-空肠吻合术(D组)124例.术后A、C两组分别有7.9％及6.0％的患者再发胆道梗阻,A、B两组分别有8.9％及8.3％的患者出现消化道梗阻,四组患者生存时间差异无统计学意义(F=1.933,P=0.123).结论 对不合并消化道梗阻的晚期胰头癌患者,胆总管空肠吻合术能有效预防术后再次胆道梗阻;预防性胃空肠吻合术能显著降低患者术后消化道梗阻发生率,而胆囊-空肠吻合术仅可在患者一般情况较差或胆管吻合条件不具备时谨慎选用.     

胰头癌腹腔镜切除的现状及展望

胰腺癌是所有常见恶性肿瘤中预后最差者,5年生存率仅约5%,根治性切除仍然是可切除胰腺癌首选的治疗方式,胰十二指肠切除术是治疗胰头癌的标准术式。近年来,腹腔镜技术在外科领域的应用越来越广泛,但腹腔镜下胰十二指肠切除术因其安全性一直存在争议,而对胰头癌实施腹腔镜下胰十二指肠切除术要比对其他疾病如壶腹部与胆总管下段占位、胰头低度恶性肿瘤等要困难的多,加之对腹腔镜技术增加恶性肿瘤播散风险的疑虑,对胰头癌实施腹腔镜切除所受质疑更多。我们结合自身开展腹腔镜治疗胰头癌的经验,对目前腹腔镜治疗胰头癌的现状进行分析。     

白藜芦醇对人胰腺癌PANC-1细胞增殖与侵袭的影响

目的 探讨白藜芦醇对人胰腺癌PANC-1细胞增殖与侵袭能力的影响.方法 实验分为空白对照组,0.1%DMSO组,白藜芦醇组(50、100、200 μmol/L).MTT法检测白藜芦醇对细胞增殖的影响;流式细胞仪检测细胞凋亡及周期变化;Transwell侵袭小室检测白藜芦醇对细胞侵袭的影响;荧光实时定量PCR和Western blot检测白藜芦醇对细胞Bax、Bcl-2、MMP-2、MMP-9表达的影响.数据以-x±s表示,多组比较采用方差分析.结果 (1)空白对照组抑制率为0;0.1%DMSO组细胞抑制率为3.25%±0.42%;白藜芦醇50 μmol/L组细胞抑制率为13.23%±1.68%;白藜芦醇100μmol/L组细胞抑制率为42.25%±3.20%;白藜芦醇200μmol/L组细胞抑制率为56.94%±5.31%.各组比较,差异有统计学意义(F=460.10,P<0.05).(2)空白对照组细胞凋亡率为0.05%±0.03%;0.1%DMSO组细胞为凋亡率为3.39%±1.77%;白藜芦醇50 μmol/L组细胞凋亡率为6.92%±1.85%;白藜芦醇100 μmol/L组细胞凋亡率为19.05%±2.01%;白藜芦醇200 μmol/L组细胞凋亡率为27.17%±6.43%.各组比较,差异有统计学意义(F=38.84,P<0.05).(3)0.1%DMSO组对细胞周期无显著影响.白藜芦醇引起PANC-1细胞G0/G1期和S期阻滞,G2/M期细胞减少.(4)空白对照组平均穿膜细胞数为61±13;0.1%DMSO组为54±13;白藜芦醇50 μmol/L组为48±15;白藜芦醇100 μmol/L组为23±6;白藜芦醇200 μmol/L组为18±7.各组比较,差异有统计学意义(F=69.08,P<0.05).(5)白藜芦醇可使PANC-1细胞Bax表达升高,Bcl-2表达下调.MMP-2、MMP-9的表达明显受到抑制,mRNA和蛋白水平变化一致.结论 白藜芦醇可明显抑制胰腺癌PANC-1细胞增殖,诱导细胞凋亡并抑制其侵袭能力.     

靶向人SIRT1基因的shRNA真核表达质粒的构建及鉴定

目的构建针对人SIRT1基因的shRNA真核表达质粒,并筛选出对胰腺癌细胞系PANC-1基因沉默效果最明显的shRNA质粒表达载体。方法针对SIRT1基因的mRNA序列设计,分别构建3个shRNA质粒表达载体和1个阴性对照质粒表达载体,经大肠杆菌扩增,酶切,PCR,测序鉴定,转染胰腺癌PANC-1细胞,实时荧光定量PCR和Western blot检测SIRT1 mRNA和蛋白的被抑制情况。结果经测序证实,成功构建SIRT1-shRNA真核表达质粒,插入的DNA片段的序列与设计序列完全一致。重组质粒转染PANC-1细胞后,SIRT1 mRNA和蛋白水平明显下调;其中以1号重组质粒效应最强。结论成功构建了携带以SIRT1为靶向的shRNA的重组质粒。其对胰腺癌PANC-1细胞SIRT1的表达具有显著抑制效应。该实验为进一步研究SIRT1的功能和肿瘤的基因治疗提供了实验基础。     

缺氧微环境对胰腺癌细胞发生上皮向间叶转化的诱导作用

Objective To investigate whether hypoxic environment can promote the metastasis of pancreatic cancer cells by inducing epithelial to mesenchymal transition (EMT). Methods The Panc-1 cells were cultured in hypoxia environment. After cultured for indicated periods, the in vitro invasive ability of Pane-1 cells was compared with normoxia group using Transwell invasion assay. The epithelial marker E-cadherin and the mesenchymal marker vimentin were assayed by Western blot. The expression of Snail, a strong activator of EMT, was detected by real-time PCR. The HIF-1 α encoding cDNA was transiently trans-fected into the Pane-1 cells, and the E-cadherin and vimentin were analyzed. Results The number of cells invading to the lower side of the membrane under hypoxia was ( 121±5 ), whereas only ( 84±3 ) in no-morxia group ( P < 0.05 ). The relative expression of E-cadherin in normoxia, hyoxia groups at 12,24,48 hwas (0.59±0.04 vs 54.00±0.05,0.45±0.10,0.36±0.03 ), and that of vimentin was (0.36±0.05, 0.41±0.04,0.48±0.06,0.58±0.05 ) ( P < 0.05 ). The relative expression of Snail mRNA was in-creased in hypoxia environment, and the difference was significant after 72 h ( P < 0.05 ). Before and after HIF-1α cDNA was transfected into the Panc-1 cells, the relative expression of E-cadherin was 0.63± 0.05, and 0.47±0.07, and that of vimentin was 0.47±0.07, and 0.32±0.04 respectively ( P < 0.05 ). Conclusion Hypoxia microenvironment can activate HIF-1α and Snail to trigger EMT of pancreatic cancer cells.     

缺氧微环境对胰腺癌细胞发生上皮向间叶转化的诱导作用

Objective To investigate whether hypoxic environment can promote the metastasis of pancreatic cancer cells by inducing epithelial to mesenchymal transition (EMT). Methods The Panc-1 cells were cultured in hypoxia environment. After cultured for indicated periods, the in vitro invasive ability of Pane-1 cells was compared with normoxia group using Transwell invasion assay. The epithelial marker E-cadherin and the mesenchymal marker vimentin were assayed by Western blot. The expression of Snail, a strong activator of EMT, was detected by real-time PCR. The HIF-1 α encoding cDNA was transiently trans-fected into the Pane-1 cells, and the E-cadherin and vimentin were analyzed. Results The number of cells invading to the lower side of the membrane under hypoxia was ( 121±5 ), whereas only ( 84±3 ) in no-morxia group ( P < 0.05 ). The relative expression of E-cadherin in normoxia, hyoxia groups at 12,24,48 hwas (0.59±0.04 vs 54.00±0.05,0.45±0.10,0.36±0.03 ), and that of vimentin was (0.36±0.05, 0.41±0.04,0.48±0.06,0.58±0.05 ) ( P < 0.05 ). The relative expression of Snail mRNA was in-creased in hypoxia environment, and the difference was significant after 72 h ( P < 0.05 ). Before and after HIF-1α cDNA was transfected into the Panc-1 cells, the relative expression of E-cadherin was 0.63± 0.05, and 0.47±0.07, and that of vimentin was 0.47±0.07, and 0.32±0.04 respectively ( P < 0.05 ). Conclusion Hypoxia microenvironment can activate HIF-1α and Snail to trigger EMT of pancreatic cancer cells.     

Etiology,Pathology, Management and Prognosis of Chronic Pancreatitis in Chinese Population: A Retrospective Study

The purpose of this study was to investigate the etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population. The clinical data of 142 patients with chronic pancreatitis were retrospectively studied. All patients were of Chinese nationality and hospitalized from January 2008 to December 2011. Their ages ranged from 14 to 76 years, with a mean of 43 years. Of 142 patients, there were 72 cases of obstructive chronic pancreatitis(50.70%), 19 cases of alcoholic chronic pancreatitis(13.38%), 14 cases of autoimmune pancreatitis(9.86%) and 37 cases of undetermined etiology(26.06%). Pathologically, the average inflammatory mass diameter was 3.8±3.3 cm, biliary obstruction occurred in 36 cases, gall stones in 70 cases, calcification in 88 cases, ductal dilatation in 61 cases, side branch dilatation in 32 cases, ductal irregularity in 10 cases, lymphocytic inflammation in 23 cases, obliterative phlebitis in 14 cases, extra pancreatic lesion in 19 cases and fibrosis in 142 cases. Location of pancreatic lesion in the region of head(n=97), neck(n=16), body(n=12), tail(n=15) and whole pancreas(n=2) influenced the choice of surgical procedures. Ninety-four patients(66.20%) received surgical treatment and 33.80% received other treatments. After operation, 80.85% of 94 patients experienced decreased pain, and 8.51% of 94 showed recovery of endocrine function but with a complication rate of 12.77%. All the operations were performed successfully. According to the pain scale of European Organization for Research and Treatment of Cancer(QLQ-C30) a decrease from 76±22 to 14±18 was observed. Etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population vary from others.     

腺病毒介导p53基因对胰腺癌细胞抑制作用的体外实验研究

目的 探讨腺病毒介导p53基因(adenovirus-mediated p53 gene,Ad-p53)对人胰腺癌细胞株PANC-1的生长抑制作用.方法 以Western Blot方法 检测加入Ad-p53后PANC-1细胞在48h 、72h p53表达情况以及未处理组细胞48h 、72h p53的表达.采用四甲基偶氮唑盐(MTT)比色实验检测不同MOI值的Ad-p53感染后,PANC-1细胞的生长情况并绘制生长曲线,用PI、annexin V-FITC双重染色并用流式细胞技术检测受感染细胞的凋亡比例.结果 Western Blot结果 显示,处理组细胞内48h 、72h p53蛋白表达明显高于相同时间点对照组细胞;MTT结果 显示不同MOI值细胞生长抑制情况不同,MOI值越高,对细胞生长的抑制程度也越高;流式检测细胞在MOI值为150时,0h、48h、72h凋亡比例分别为(7.5±0.8)%、(22.5±2.3)%和(34.4±2.7)%,对照组细胞为(5.8±0.4)%、(8.3±1.7)%和(9.7±2.1)%.结论 Ad-p53能有效感染胰腺癌细胞,导致胰腺癌细胞凋亡.     

Surgical treatment of chronic pancreatitis in young patients

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography （ERCP） intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were ana- lyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of stea- torrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP in- tervention on delaying the progression of the disease and relieving the symptoms.     

缺氧微环境对胰腺癌细胞发生上皮向间叶转化的诱导作用

Objective To investigate whether hypoxic environment can promote the metastasis of pancreatic cancer cells by inducing epithelial to mesenchymal transition (EMT). Methods The Panc-1 cells were cultured in hypoxia environment. After cultured for indicated periods, the in vitro invasive ability of Pane-1 cells was compared with normoxia group using Transwell invasion assay. The epithelial marker E-cadherin and the mesenchymal marker vimentin were assayed by Western blot. The expression of Snail, a strong activator of EMT, was detected by real-time PCR. The HIF-1 α encoding cDNA was transiently trans-fected into the Pane-1 cells, and the E-cadherin and vimentin were analyzed. Results The number of cells invading to the lower side of the membrane under hypoxia was ( 121±5 ), whereas only ( 84±3 ) in no-morxia group ( P < 0.05 ). The relative expression of E-cadherin in normoxia, hyoxia groups at 12,24,48 hwas (0.59±0.04 vs 54.00±0.05,0.45±0.10,0.36±0.03 ), and that of vimentin was (0.36±0.05, 0.41±0.04,0.48±0.06,0.58±0.05 ) ( P < 0.05 ). The relative expression of Snail mRNA was in-creased in hypoxia environment, and the difference was significant after 72 h ( P < 0.05 ). Before and after HIF-1α cDNA was transfected into the Panc-1 cells, the relative expression of E-cadherin was 0.63± 0.05, and 0.47±0.07, and that of vimentin was 0.47±0.07, and 0.32±0.04 respectively ( P < 0.05 ). Conclusion Hypoxia microenvironment can activate HIF-1α and Snail to trigger EMT of pancreatic cancer cells.