胰腺癌细胞株化疗敏感性与胸苷酸合成酶蛋白表达的关系

目的 探讨胸苷酸合成酶(TS)在胰腺癌细胞中的表达及其蛋白表达水平与胰腺癌细胞化疗敏感性的关系.方法 利用CCK-8试剂盒检测7株胰腺癌细胞(AsPC-1、BxPC-3、Capan-1、SU86.86、T3M4、PANC-1与COLO357)对5-氟脲嘧啶(5-Fu)的化疗敏感性,应用Western blot检测其TS蛋白表达.结果 细胞株AsPC-1、BxPC-3、Capan-1、SU86.86、T3M4中TS蛋白表达水平低(低TS表达组),而PANC-1与COLO357中TS蛋白表达水平较高(高TS表达组),低TS表达组的细胞在低中高3个不同药物浓度下对5-Fu抑制率分别为0.683±0.131、0.769±0.114与0.836±0.094.同浓度下高TS表达组的细胞对5-Fu抑制率分别为0.370±0.157、0.548±0.018与0.638±0.020.TS蛋白表达低的细胞对5-Fu化疗敏感性明显高于TS表达高的细胞,差异有统计学意义(P＜0.05).结论 在胰腺癌细胞株中,TS蛋白表达水平与其对5-Fu的化疗敏感性密切相关.     

胰腺癌——胰腺癌的辅助化疗

胰腺癌的发病率近年来在世界范围内有逐年增高的趋势，虽然手术是治疗胰腺癌的根本手段，但目前胰腺癌手术切除率仅为10％～15％，因此围手术期化疗、放疗等综合治疗尤为重要。目前使用的化疗药物中5-氟脲嘧啶(5-Fu)和丝裂霉素(MMC)的抗胰腺癌作用较为肯定，广为临床应用。目前各化疗方案仍主要以5一Fu为基础，最常用的方案有：5-Fu 阿霉素(ADM) MMC；5-Fu 顺铂(DDP)；5-Fu 健择(GEM)等。     

抗表皮生长因子受体单克隆抗体对人胰腺癌裸鼠模型的化疗增敏作用

目的 观察鼠抗人EGFR单克隆抗体联合5-氟尿嘧啶(5-Fu)、健择对裸鼠胰腺癌化疗效果的影响.方法 将人胰腺癌肿瘤细胞悬液注射于裸鼠背部皮下,建立肿瘤模型.干预组腹腔分别注射鼠抗人EGFR单克隆抗体(MMAb-2)、5-Fu、健择及其配伍联合用药;对照组腹腔注射生理盐水.干预4周后处死裸鼠,切取肿瘤,测量瘤体积,取肿瘤组织送病理学检查;免疫组化法检测肿瘤组织中bax、bcl-2的表达;免疫荧光法检测肿瘤组织中细胞凋亡指数,评价干预效果.结果 5-Fu、健择联合应用MMAb-2对裸鼠胰腺癌细胞增殖的抑制作用明显增强(P＜0.05),且能提高裸鼠胰腺癌细胞凋亡率(P＜0.05).结论 抗EGFR单克隆抗体能有效提高化疗药物5-Fu、健择对裸鼠胰腺癌细胞抑制作用的敏感性,可明显提高胰腺癌的化疗效果.     

DPC4基因转染对胰腺癌细胞化疗敏感性的影响

目的 运用基因转染技术,观察DPC4基因转染对胰腺癌细胞化疗敏感性的影响.方法 构建表达DPC4基因的逆转录病毒载体,转染胰腺癌细胞BxPC-3,获取稳定表达DPC4的子代胰腺癌的细胞株BxPC-3/DPC4.观察5-Fu、吉西他滨(作用72 h)对胰腺癌细胞的抑制作用.同时应用半定量RT-PCR检测胰腺癌细胞内Mdr-1、Chk1基因的表达情况.结果 DPC4基因转染并稳定表达后,BxPC-3细胞对5-Fu、吉西他滨的IC50浓度(72 h)分别降低了1倍左右.同一浓度下,5-Fu、吉西他滨联合DPC4基因转染对BxPC-3细胞的体外抑制作用也明显增强,癌细胞内的Mdr-1、Chk1基因的mRNA表达明显下调.显示单独使用pLXSN/DPC4、5-Fu、吉西他滨,均能在体外抑制胰腺癌细胞的生长,但pLXSN/DPC4联合化疗药物,对癌细胞生长的抑制作用更为明显.结论 DPC4基因转染能够提高胰腺癌细胞对化疗药物的敏感性,其机制可能是通过下调Mdr-1、Chkl的表达来实现的.     

DPC4基因转染对胰腺癌细胞化疗敏感性的影响

目的 运用基因转染技术,观察DPC4基因转染对胰腺癌细胞化疗敏感性的影响.方法 构建表达DPC4基因的逆转录病毒载体,转染胰腺癌细胞BxPC-3,获取稳定表达DPC4的子代胰腺癌的细胞株BxPC-3/DPC4.观察5-Fu、吉西他滨(作用72 h)对胰腺癌细胞的抑制作用.同时应用半定量RT-PCR检测胰腺癌细胞内Mdr-1、Chk1基因的表达情况.结果 DPC4基因转染并稳定表达后,BxPC-3细胞对5-Fu、吉西他滨的IC50浓度(72 h)分别降低了1倍左右.同一浓度下,5-Fu、吉西他滨联合DPC4基因转染对BxPC-3细胞的体外抑制作用也明显增强,癌细胞内的Mdr-1、Chk1基因的mRNA表达明显下调.显示单独使用pLXSN/DPC4、5-Fu、吉西他滨,均能在体外抑制胰腺癌细胞的生长,但pLXSN/DPC4联合化疗药物,对癌细胞生长的抑制作用更为明显.结论 DPC4基因转染能够提高胰腺癌细胞对化疗药物的敏感性,其机制可能是通过下调Mdr-1、Chkl的表达来实现的.     

DPC4基因转染对胰腺癌细胞化疗敏感性的影响

目的 运用基因转染技术,观察DPC4基因转染对胰腺癌细胞化疗敏感性的影响.方法 构建表达DPC4基因的逆转录病毒载体,转染胰腺癌细胞BxPC-3,获取稳定表达DPC4的子代胰腺癌的细胞株BxPC-3/DPC4.观察5-Fu、吉西他滨(作用72 h)对胰腺癌细胞的抑制作用.同时应用半定量RT-PCR检测胰腺癌细胞内Mdr-1、Chk1基因的表达情况.结果 DPC4基因转染并稳定表达后,BxPC-3细胞对5-Fu、吉西他滨的IC50浓度(72 h)分别降低了1倍左右.同一浓度下,5-Fu、吉西他滨联合DPC4基因转染对BxPC-3细胞的体外抑制作用也明显增强,癌细胞内的Mdr-1、Chk1基因的mRNA表达明显下调.显示单独使用pLXSN/DPC4、5-Fu、吉西他滨,均能在体外抑制胰腺癌细胞的生长,但pLXSN/DPC4联合化疗药物,对癌细胞生长的抑制作用更为明显.结论 DPC4基因转染能够提高胰腺癌细胞对化疗药物的敏感性,其机制可能是通过下调Mdr-1、Chkl的表达来实现的.     

5-氟尿嘧啶缓释剂瘤内注射治疗胰腺癌的实验研究和临床研究

目的 观察5-氟尿嘧啶(5-Fu)缓释剂对荷胰腺癌裸鼠肿瘤细胞及胰腺癌患者血清肿瘤标记物和细胞免疫的影响。方法 (1)5-Fu缓释剂的体外释放实验和体外抑瘤实验：测定浸出液药物的浓度，计算释放量；检测其浸出液对人胰腺癌细胞株PC3的抑制作用：(2)将荷胰腺癌细胞株Pc3裸鼠60只，随机分成静脉对照组(A组)、5-Fu静注组(B组)、基质植入组(C组)、大剂量5-Fu缓释剂植入组(D组)和小剂量5-FU缓释利植入组(E组)：治疗前及治疗后l4d测肿瘤大小。治疗2周后观察肿瘤组织学变化：免疫组化法测定bcl-2和Bax的蛋白表达水平；TUNEL法检测凋亡指数(Al)。(3)手术探查不能切除之胰腺癌69例随机分成3组：将5-FU缓释剂瘤内植入治疗组(治疗组)、术后行5-FU静脉化疗组(化疗组)和对照组。分别于术前1d和术后第14天采血，测定各组血清中NK细胞，T细胞亚群和CEA，CA50，CA19-9，CA125，CA242血清肿瘤标记物水平。结果 (1)5mg 5-FU缓释剂第1天释放量最大，为0．85mg，第3天为0．45mg，其后在0．25mg水平维持稳定的缓慢释放；释放时间长达l4d以上。(2)5-Fu缓释剂第1天的浸出液对人胰腺癌细胞株PC-3的抑制率达60．27％，第3天为34．25％，以后稳定在25．00％左右。5-Fu缓释剂瘤内注射治疗组裸鼠移植瘤生长速度减慢，bcl-2基因表达明显低于其他各组，而Bax基因表达明显高于其他各组，肿瘤细胞的Al明显高于其他各组。D组和E组肿瘤组织中炎症反应和血管内膜增厚程度明显高于其他各组。术后治疗组CD4 ／CD8 和NK细胞水平高于化疗组，而血清中E述5种肿瘤标记物低于对照组和化疗组。结论 5-Fu缓释剂能在2周内在体外较稳定地持续释放，对人胰腺癌细胞株PC3有持续抑制作用。该剂瘤内注射可明显抑制荷胰腺癌瘤裸鼠瘤体的生长，其作用机制与药物在肿瘤组织中引起的炎症反应和血管内膜增厚等因素有关；并可能与诱导肿瘤细胞的凋亡有关。该剂植入患者胰腺癌实体内，能明显降低5种血清肿瘤标记物水平，同时对患者的细胞免疫功能影响较小，5-Fu缓释剂可望成为治疗不能切除之胰腺癌的较好的制剂。     

全身性热化疗治疗晚期胰腺癌的临床观察

目的　探讨全身性热化疗治疗晚期胰腺癌的效果,以及化疗药物在此过程中代谢的变化。方法　将42例晚期胰腺癌病人分成2组,分别接受全身性热化疗和单纯化疗后,比较二者的疗效、不良反应及5 FU的药代动力学指标。结果　实验组具有较好的疗效,不良反应的发生率随之提高,其体内化疗药物的相对浓度升高。结论　全身性热化疗是一种有效的治疗晚期胰腺癌的方法,其机理与化疗药物在体内有效浓度的增加有密切的关系。     

区域性动脉灌注化疗对胰腺癌淋巴结转移的影响

区域性动脉灌注化疗(RAIC)作为胰腺癌综合治疗的有效手段,不仅可提高胰腺癌局部的药物浓度,降低全身不良反应,提高耐受性,从而保证足够的给药剂量,还有可能提高胰腺区域内淋巴结内的药物浓度,减少淋巴结转移的发生,但胰腺癌RAIC作用机制和对胰腺癌淋巴结转移的影响有待进一步研究.     

5-氟尿嘧啶腹腔内缓释化疗的家兔门静脉血药浓度测定

目的:测定5-氟尿嘧啶(5-FU)腹腔内缓释化疗后家兔门静脉血中的药物浓度。方法:以家兔为模型分别行5-FU静脉化疗、腹腔化疗和缓释化疗,用高效液相色谱法测定门静脉血中的5-FU浓度。根据各组门静脉血浓度-时间曲线,计算曲线下面积(AUC)。结果:缓释化疗组家兔门静脉血药浓度最高值出现在给药后2h左右,达0.82μg/ml,但持续时间较长,且随时间延长门静脉血药浓度变化较小,在给药16d后仍可测出。腹腔化疗组给药后15min门静脉血5-FU浓度达20.67μg/ml,但持续时间短,24h后浓度极低。AUC静脉组=21.57(μg/ml·h),AUC腹腔组=42.85(μg/ml·h),AUC缓释组=70.08(μg/ml·h)。结论:5-FU腹腔内缓释化疗延长了门静脉血中有效浓度的持续时间,增强了对肝脏的化疗效果。     

联合门静脉胰腺癌切除术的安全性及疗效评价

目的对联合门静脉切除胰腺癌的可行性进行分析,为胰腺癌的治疗提供新途径。方法以行联合门静脉胰腺癌切除术的病变累及门静脉胰腺癌35例患者为观察组,20例同期行姑息旁路术的同类患者为对照1组,同期病灶未累及门静脉行标准根治手术的20例胰腺癌患者为对照2组,观察3组的手术效果及并发症情况。结果采用联合门静脉切除胰腺癌患者的手术后并发症发生率为25％,与对照2组的23．5％相比差异无统计学意义（P〉0．05）;对患者采用寿命表进行计算,观察组患者术后1、3、5年的生存率分别为78．5％,28．8％,9．58％,与对照2组的80．5％,30．9％以及10．23％的差异无统计学意义（均P〉0．05）,但显著高于对照1组的48．5％,11．5％,1．5％（均P〈0．05）。结论对于病变已累及门静脉的胰腺癌采用联合门静脉胰腺癌切除术,可有效提高胰腺癌手术切除率,有效延长患者寿命,改善患者预后。     

Oxytetracycline concentration of bile after systemic and regional administration.

Oxytetracycline concentration of bile was determined after systemic and regional (intraportal) infusions. A single dose of 3.60 mg/kg of oxytetracycline was infused into the brachial vein of 8 dogs and into the portal vein of 9 dogs in 120 minutes. Bile samples taken at 30, 60, 120 and 180 minutes showed that the liver excretes into the bile higher concentrations of oxytetracycline in the case of intraportal infusion. The results suggest that treatment of inflammatory diseases of bacterial origin of the biliary tree might be more effective when the antibiotic is applied in intraportal infusion.     

Survival benefits of portal vein resection for pancreatic cancer

BACKGROUND: The efficacy of portal vein resection for pancreatic cancer is controversial. METHODS: Eighty-one consecutive patients with pancreatic cancer undergoing surgical resection were retrospectively analyzed. The clinicopathological findings and relationship between portal vein resection and survival were investigated. RESULTS: Thirty-three patients with pancreatic cancer underwent pancreatic resection with portal vein resection. Histological examination revealed that 17 patients had definite invasion to the portal vein (group 1) and 16 patients had no invasion (group 2). Forty-eight patients with pancreatic cancer underwent pancreatic resection without portal vein resection (group 3). There were no significant differences in survival rates (P = 0.437) between patients with portal vein resection and patients without portal vein resection. However, patients in group 1 had a significantly (P = 0.021) worse prognosis as compared with those in group 2. Despite aggressive surgical resection, the surgical margin was positive in 35% of patients in group 1 as compared with 13% of patients in group 2 and 21% of patients in group 3. CONCLUSIONS: Patients undergoing portal vein resection for pancreatic cancer had a prognosis similar to patients without portal vein resection. Negative microscopic invasion to the portal vein was significantly associated with improved survival.     

Intraportal endovascular ultrasonography in the diagnosis of portal vein invasion by pancreatobiliary carcinoma.

OBJECTIVE: The purpose of this study was to determine the value of intraportal endovascular ultrasonography (IPEUS) in the diagnosis of portal vein invasion by pancreatobiliary carcinoma. The authors reported their experiences with this new technique and compared it with conventional imaging technologies, such as portography and computed tomography (CT). SUMMARY BACKGROUND DATA: Pancreatobiliary carcinoma often invades the portal vein. Observation of the echogenic band of the portal vein wall by means of a high-frequency, high-resolution intravascular ultrasound catheter allows for the accurate diagnosis of the portal vein invasion. METHODS: A prospective study of 30 consecutive patients with pancreatobiliary carcinoma (16 pancreatic carcinomas, 8 bile duct carcinomas and 6 gallbladder carcinomas) was performed. In 23 cases IPEUS was performed intraoperatively from the superior mesenteric venous route with an 8 French, 20 MHz intravascular ultrasound catheter. In 7 cases IPEUS was performed before surgery from the percutaneous transhepatic route with a 6 French, 20 MHz intravascular ultrasound catheter. The finding of IPEUS was confirmed by pathologic examination of resected specimens and surgical exploration. The results of IPEUS were compared to those of portography and CT. RESULTS: Intraportal endovascular ultrasonography visualized the portal vein wall as an echogenic band with a thickness of 0.5 mm to 1.0 mm. The diagnostic criterion of portal vein invasion was destruction of this echogenic band. Portal vein invasion was found in 15 of 30 cases. Vascular invasion was confirmed by pathologic examination of resected specimens in 10 patients and operative findings in 5. The sensitivity, specificity, and overall accuracy of IPEUS for diagnosis of portal vein invasion was 100%, 93.3%, and 96.7%, respectively. The values were 80%, 67.7%, and 73.3% for portography and 53.3%, 80%, and 66.7%, respectively, for CT. CONCLUSIONS: Intraportal endovascular ultrasonography provided precise information about the relationship between the pancreatobiliary tumor and the portal vein wall. It was capable of accurately detecting or excluding early invasion of the portal vein wall by pancreatobiliary carcinoma.     

Intraoperative irradiation in resected carcinoma of the pancreas and portal vein.

We developed a new method that provides a wide retroperitoneal irradiation field for patients with cancer of the pancreas in whom a long invaded portal vein is resected during intraoperative radiotherapy. This method involves the use of a venous bypass with a heparinized catheter that does not require systemic anticoagulation or a pump. Anastomosis is completed after intraoperative radiotherapy. Mean (+/- SD) duration of the venous bypass was 3.0 +/- 1.5 hours (range, 2.0 to 6.6 hours), during which no abnormalities of systemic blood pressure, heart rate, and electrocardiographic activity and no signs of congestion of the intestines were observed. Eleven patients were treated with this method, and no complications ascribed to intraoperative radiotherapy or venous bypass were found. We recommend this method as safe and simple for expanding the irradiation field for intraoperative radiotherapy in patients with pancreatic cancer in whom the portal vein is resected.     

ZT胶栓塞兔门静脉的可行性研究

目的探讨不同浓度ZT胶栓塞兔门静脉的可行性及其机制。方法24只新西兰白兔分为三组,A组、B组分别为ZT胶:碘油1∶1、1∶2混合物组,对照组C组为碘油组,数字减影血管造影(DSA)下对门静脉左支进行栓塞,术后检测门静脉压力、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)的变化,术后30dDSA下门静脉造影。结果门静脉栓塞(PVE)后门静脉压力、ALT、AST呈一过性升高。术后5min门静脉造影,A组表现为门静脉大分支栓塞,B组、C组为周围性栓塞。术后30d门静脉造影,A组、B组呈永久性栓塞。结论不同浓度ZT胶可选择性栓塞到门静脉各级分支,并造成永久性栓塞。     

Portal vein resection with a new antithrombogenic catheter

Curative resection of pancreatic and hepatobiliary tumors is rarely possible because of local invasion, especially into the portal vein. We developed a new antithrombogenic catheter using a heparinized hydrophilic polymer to allow portal vein bypass during resection of tumors invading the portal vein. Pancreatectomy or hepatectomy accompanied by portal vein resection was performed for pancreatic or hepatobiliary cancer, with an intraoperative shunt from the superior mesenteric vein to the femoral vein or from the superior mesenteric vein to the intrahepatic portal vein through the umbilical vein or the hepatic hilar portal vein. Use of the shunt prevented stasis in the superior mesenteric vein and hepatic ischemia even during prolonged occlusion of the portal vein, and portal vein resection was performed in 81 patients with hepatobiliary and pancreatic disease with greater safety and ease.     

Combined regional thrombolysis and surgical thrombectomy for treatment of iliofemoral vein thrombosis

OBJECTIVE: In at least half of patients with iliofemoral deep vein thrombosis post-thrombotic syndrome develops when only anticoagulant therapy is given. We combined thrombolysis, applied under ischemic conditions,with surgical thrombectomy to restore patency and valve function. The technique and the short-term and long-term results in 2 patient series are reported. METHODS: A catheter was inserted into a foot vein of the thrombosed leg, and the limb was excluded from the circulation with a pneumatic cuff placed on the thigh with the patient under general anesthesia. Urokinase (0.5 million-3 million IU) and heparin were infused and allowed to act for 30 minutes while the pelvic axis was cleared with a Fogarty catheter through an inguinal venotomy. The external iliac vein was then clamped and the cuff removed. Thrombi that detached from the wall were flushed out with reactive hyperemia and squeezed out with manual leg compression. The blood was retrieved, washed, and transfused back into the patient. Various additional procedures were performed to secure outflow. Two patient series are reported: 1 with 12 consecutive patients and 1 with 21 patients who were successfully treated 6 to 10 years previously. Follow-up data were obtained for all patients after 1 year and for 18 of 21 patients after 6 to 10 years. Patency and valve function were assessed with duplex scanning or venography. Studies of blood coagulation and the kinetics of urokinase were performed in 5 additional patients. RESULTS: Vein patency and valve function were restored in all consecutive patients. At 1 year none of the 33 patients had had recurrence, and none showed clinical signs of post-thrombotic syndrome. At 6 to 10 years 3 of 18 patients had experienced another venous thromboembolism, but none in the treated leg. Sixteen legs were asymptomatic without compression therapy, and 2 had venous claudication. Coagulation studies showed a trace concentration of urokinase and a mild decrease in fibrinogen in the systemic circulation. The concentration of urokinase in blood collected from the treated leg was only 1% of that infused. CONCLUSION: Regional thrombolysis combined with surgical thrombectomy is relatively easy to perform and seems safe. Vein patency and valve function were restored, and post-thrombotic syndrome was prevented. Additional procedures to overcome pelvic vein obstructions were required in 11 of 33 patients (33%). The procedure should be tested against standard anticoagulation therapy in patients with acute iliofemoral thrombosis.     

Pancreatectomy combined with superior mesenteric-portal vein resection for adenocarcinoma in pancreas

The aims of this study were to investigate morbidity, mortality, and survival of patients with ductal adenocarcinoma of the pancreas who underwent pancreatectomy without (group 1) or with (group 2) en bloc portal vein resection and to study the degree of carcinoma invasion of the portal vein in group 2. The medical records of 46 and 28 patients in groups 1 and 2, respectively, were reviewed. In addition, the degree of invasion of the wall of the portal vein was categorized histologically into three types: type I, transmural invasion involving the intima; type II, invasion of the wall of the vein without intimal involvement; and type III, compression of the wall of the vein by surrounding carcinoma without true invasion. The morbidity and mortality in group 1 (26% and 4%) were not different from those in group 2 (32% and 4%). Similarly, there was no difference in survival between the two groups. Survival tended to vary directly with the depth of invasion of the wall of the portal vein: type I 6.8 ± 1.9 months; type II 15.3 ± 6.4 months; type III 20.6 ± 13.0 months. These findings suggest that en bloc resection of the pancreas and the portal vein does not increase mortality and morbidity after pancreatectomy; survival after en bloc resection was similar to that of patients not requiring portal vein resection. Combined resection of the pancreas with the portal vein could be an option in the treatment of pancreatic cancer with direct invasion of the portal vein.     

Changes of local and systemic hemorheological properties in intestinal ischemia‐reperfusion injury in the rat model

The aim of this study was to investigate intestinal ischemia‐reperfusion and its local and systemic hemorheological relations in the rat. Ten anaesthetized female CD outbred rats were equally divided into 2 experimental groups. (1) Ischemia‐reperfusion (I/R): the superior mesenterial artery was clipped for 30 minutes. After removing the clip, 60 minutes of the reperfusion was observed before extermination. Blood samples were taken from the caudal caval vein and from the portal vein before ischemia, 1 minute before and after clip removal, and at the 15th, 30th, and 60th minutes of the reperfusion. (2) Sham operation: median laparotomy and blood sampling were done according to the timing as in I/R group. Hematological parameters, red blood cell aggregation, and deformability were determined. Leukocyte count and mean volume of erythrocytes increased slightly but continuously in portal venous samples during the reperfusion period. Red blood cell aggregation values were higher in portal blood by the end of ischemia, and then became elevated further comparing to the caval venous blood. Both in caval and portal venous samples of I/R group red blood cell deformability significantly worsened during the experimental period compared to its base and Sham group. In portal blood red blood cell deformability was impaired more than in caval vein samples. Histology showed denuded villi, dilated capillaries, and the inflammatory cells were increased after a 30 minutes ischemia. In conclusion, intestinal ischemia‐reperfusion causes changes in erythrocyte deformability and aggregation, showing local versus systemic differences in venous blood during the first hour of reperfusion. © 2009 Wiley‐Liss, Inc. Microsurgery, 2010.