胰腺导管腺癌、慢性胰腺炎及正常胰腺组织中胰腺上皮内瘤变的比较研究

目的探讨胰腺导管腺癌、慢性胰腺炎和正常胰腺组织中各级别PanIN的发生率以及与临床病理学参数间的关系。方法回顾性研究长海医院2001年1月-2003年12月间外科切除和同期尸检获得的250例胰腺标本中PanIN的发生情况,并联系临床病理指标进行相关分析。结果250例胰腺标本中,有156例存在PanIN病变,发生率62.4％。其中,胰腺导管腺癌、慢性胰腺炎和正常胰腺组织中PanIN的发生率分别为75.6％、46.0％和30.0％,胰腺导管腺癌PanIN发生率明显高于慢性胰腺炎及正常胰腺组织(P<0.01);慢性胰腺炎中高级别PanIN发生率明显高于正常胰腺组织(P<0.05)。PanIN-3仅在胰腺导管腺癌和慢性胰腺炎中见到。胰腺导管腺癌中,有烟酒嗜好和(或)糖尿病者高级别PanIN的发生率53.7％,明显高于对照组29.4％(P<0.01)。PanIN的发生率以61-70岁年龄组为最高。结论胰腺导管腺癌、慢性胰腺炎和正常胰腺组织中PanIN的发生率逐渐增加,程度逐渐加重,支持胰腺癌发生的分子模型。     

胰腺癌513例神经浸润的病理分析

目的 探讨胰腺癌神经浸润的特征及其与其他临床病理参数之间的关系.方法 光镜下观察491例胰腺导管腺癌、22例其他胰腺恶性肿瘤、41例胰腺良性病变和21例慢性胰腺炎组织中的神经浸润状况,分析其与其他病理学指标的相关性.结果 胰腺导管腺癌的神经浸润率为74%,显著高于其他类型恶性肿瘤的23%(P＜0.01).导管腺癌癌细胞通常穿越外周神经中膜到达内部的神经纤维束,有的甚至横断整根神经纤维.但神经浸润与导管腺癌的分化程度无关.52%的胰腺导管腺癌癌旁组织呈慢性炎症改变,且程度严重,远高于其他类型胰腺癌(14%)及胰腺良性病变(15%)的慢性炎症发生率(P＜0.01).胰腺导管腺癌淋巴细胞浸润神经的发生率为65%,远高于其他恶性肿瘤的36%和胰腺良性病变的22%(P＜0.01).胰腺导管腺癌的神经浸润与癌旁慢性胰腺炎症以及淋巴细胞浸润神经均相关,但与淋巴结转移无关.结论 神经浸润是胰腺导管腺癌特征性的生物学行为之一.     

E-钙黏附素和β-连环素在胰腺上皮内瘤变和胰腺癌组织中的表达

目的:探讨胰腺上皮内瘤变PanIN和胰腺癌组织中E-钙黏附素(E-Cad)和β-连环素(β-Cat)异常表达的意义.方法:回顾性研究长海医院2001-01/2003-12间外科切除和同期尸检的156例胰腺标本,并构建了组织芯片,其中含有129灶PanIN-1A,104灶PanIN-1B,22灶PanIN-2,11灶PanIN-3和121例导管腺癌和相应癌旁组织.用EnVision免疫组化技术检测上述病变组织中E-Cad和β-Cat的表达变化,并结合临床病理资料进行相关分析.结果:导管腺癌中E-Cad异常表达率明显高于PanINs和正常导管(64.5%,32.3%,0%),且与胰腺癌的分化程度、淋巴结转移和神经浸润密切相关(P＜0.05).PanINs和导管腺癌中E-Cad胞质表达较正常导管明显增加.β-Cat的异常表达与胰腺癌淋巴结转移和神经浸润有明显相关性(P＜0.05).高级别PanINs和导管腺癌中β-Cat胞质和胞核的表达率明显高于低级别PanINs和正常导管(P＜0.05).PanINs和导管腺癌中E-Cad和β-Cat表达间呈正相关性(P＜0.01,P＜0.05).结论:胰腺癌和PanINs中E-Cad和β-Cat的异常改变提示他们不仅与胰腺癌的生物学行为和预后有关,而且也参与了胰腺癌的发生.     

胰腺癌组织中GABA和GAD的表达

目的 研究γ-氨基丁酸(GABA)、谷氨酸脱羧酶(GAD)65和GAD67在胰腺癌组织中的表达及基意义.方法 收集34例胰腺癌、12例慢性胰腺炎和10例正常胰腺组织标本,应用ABC 法检测三者GABA、GAD65和GAD67的表达.结果 胰腺癌GABA、GAD65和GAD67表达染色强度分值(2.41±0.49,2.09±0.29,2.10±0.30)及阳性细胞数分值(4.34±0.77,3.0±0.87,3.86±0.76)明显高于慢性胰腺炎(1.55±0.83,1.22±0.63,1.44±0.68;2.77±0.78,2.35±0.83,1.88±0.78)和正常胰腺组织(1.17±0.69,0.83±0.37,1.25±0.62;1.92±0.83),1.60±1.11,1.41±0.74),差异显著(P＜0.05或P＜0.01).慢性胰腺炎GABA和GAD65的表达明显高于正常胰腺组织(P＜0.05).低分化腺癌GABA和GAD65明显高于高分化腺癌(P＜0.05).胰腺癌的GABA与GAD65表达呈正相关(r=0.67,P＜0.01).结论 GABA、GAD65T和GAD67的表达与胰腺癌的发生、发展及生物学行为有密切关系,可能是潜在的胰腺炎标志物之一.     

Midkine在胰腺癌组织中的表达及临床意义

目的 探讨人胰腺癌组织中Midkine(MK)的表达及其与肿瘤细胞增殖的关系和意义.方法 免疫组织化学SP法检测49例胰腺癌、13例慢性胰腺炎及15例正常胰腺组织中MK和Ki67蛋白的表达.结果 胰腺癌组织中MK和Ki67均高表达,阳性率分别为77.1 %(35/49)、81.6%(40/49),二者的表达都与肿瘤组织学分级、临床分期和淋巴结转移有关(P ＜0.05).胰腺癌组织中Ki67表达显著高于慢性胰腺炎(3/13,P ＜0.01)和正常胰腺组织(0/15,P ＜0.01).慢性胰腺炎和正常胰腺组织中未见MK阳性表达.胰腺癌组织中MK蛋白的表达与Ki67的表达呈正相关(r ＝ 0.4,P ＜0.05).结论 检测MK蛋白对于胰腺癌的诊断及与慢性胰腺炎的鉴别诊断具有参考价值.MK在胰腺癌中高表达可能与肿瘤的发生发展及细胞增殖密切相关.     

脑源性神经营养因子在胰腺导管腺癌中的表达及其临床意义

目的 探讨脑源性神经营养因子（BDNF）在胰腺导管腺癌中的表达及其临床意义.方法 采用免疫组化SP法检测46例胰腺导管腺癌、38例胰腺良性疾病及20例正常胰腺组织中的BDNF表达,应用蛋白质印迹法和RT-PCR法检测BDNF蛋白及mRNA表达量,并分析BDNF表达与胰腺导管腺癌临床病理参数的相关性.结果 胰腺导管腺癌组织中BDNF阳性表达率为52.2％ （24/46）,胰腺良性疾病组织为7.8％ （3/38）,而正常胰腺组织未见BDNF阳性表达.正常胰腺、胰腺良性疾病、胰腺癌组织中BDNF蛋白表达量分别为0.38±0.01、0.56±0.01、0.97±0.01,mRNA表达量分别为0.85±0.14、1.67±0.21、3.45±0.67,胰腺癌和胰腺良性疾病的表达量均显著高于正常胰腺,胰腺癌又显著高于胰腺良性疾病,差异均具有统计学意义（P值均＜0.01）.BDNF阳性表达与胰腺导管腺癌的神经浸润、淋巴结转移存在相关性（P值均＜0.05）,而与患者性别、年龄及肿瘤大小、部位、分化程度等无相关性.结论 BDNF参与胰腺癌的发生、发展,并可能与胰腺癌患者预后有关.     

新生淋巴管在胰腺导管腺癌周围神经丛微转移过程中的机制研究

目的 研究新生淋巴管在胰腺导管腺癌周围神经丛微转移过程中的作用与机制.方法 收集2005年9月至2006年10月长海医院行胰腺癌扩大根治术的30例胰腺导管腺癌患者的临床资料,术中采集胰腺肿瘤、癌旁、胆管下段、胰尾、肠系膜上动脉(SMA)旁组织(含胰周神经丛)以及区域淋巴结标本.常规病理检查,采用双重免疫组化方法检测毛细淋巴管,计算淋巴管密度(LVD).结果 胰内和(或)胰周神经丛浸润25例(83.3%),其中胰内合并胰周神经丛浸润20例,单纯胰内神经浸润5例,无单纯胰周神经丛浸润病例.神经浸润与患者年龄、性别、淋巴结转移、肿瘤大小、肿瘤部位无明显相关性(P>0.05),但与JPS临床分期相关(P<0.05).癌组织内的平均LVD为每视野(4.2±3.4)个,显著少于癌旁的(11.3±6.9)个及正常胰腺组织的(10.8±4.4)个(P<0.01),正常胰腺组织与癌旁组织平均LVD值差异无统计学意义.18例胰腺癌患者在非癌组织清晰可见肿瘤浸润淋巴管,而且胰周神经丛浸润与淋巴管肿瘤浸润间亦有明显相关性(P<0.05).结论 胰腺导管腺癌周围神经丛浸润的发生率较高,神经浸润与JPS临床分期、淋巴管浸润有明显相关性,提示胰腺导管腺癌存在通过新生淋巴管途径扩散转移的可能性.     

胰腺上皮内瘤变研究进展

胰腺癌恶性程度高,早期诊断困难,预后差.胰腺上皮内瘤变（PanIN）是胰腺导管腺癌（PDAC）最常见的非侵袭性前体病变.深入认识PanIN,能为在进展至侵袭性PDAC之前早期检出PanIN以及寻找有效治疗手段提供依据.本文对PanIN的临床、病理特征和遗传学改变作一综述.     

纤维化胰腺组织中TGF-β1、Smad3、Smad7的表达及意义

目的检测人纤维化胰腺组织中TGF-β1,Smad3和Smad7蛋白的表达,探讨他们在胰腺组织纤维化发生中的意义.方法采用免疫组织化学SP法检测34例纤维化胰腺组织标本中TGF-β1,Smad3和Smad7蛋白的表达,并以同期15例正常胰腺组织作对照.结果TGF-β1,Smad3在纤维化胰腺组织中的表达(79.4%,64.7%)明显高于正常胰腺组织(6.7%,20.0%)(P＜0.01),Smad7在纤维化胰腺组织中的表达(26.5%)则低于正常胰腺组织(73.3%)(P＜0.01),纤维化胰腺组织中TGF-β1的表达与Smad3的表达呈正相关(r=0.385,(P＜0.05)而与Smad7的表达呈负相关(r=-0.519,P＜0.01).结论TGF-β1,Smad3和Smad7在胰腺纤维化形成过程中起不同的介导作用,通过靶向性阻断TGF-β1、Smad3信号和(或)加强Smad7信号可能成为治疗胰腺纤维化的新手段.     

慢性胰腺炎、胰腺癌组织中血管生成与ras基因产物p21蛋白表达的关系及意义

目的探讨慢性胰腺炎(chronic pancreatitis,CP)、胰腺癌(pancreatic adenocarcinoma,PAC)组织中血管生成与ras基因产物p21蛋白表达的关系及在慢性胰腺炎、胰腺癌演化过程中所起的重要作用.方法应用免疫组化Envision方法检测7例正常胰腺、24例慢性胰腺炎、24例胰腺癌组织中CD34、p21ras的表达.对CD34阳性血管进行微血管密度(microvessel density,MVD)计数.结果CD34抗体染色大部分毛细微血管和单个内皮细胞.最高微血管染色区域(hotspot)几乎总是在肿瘤与周围组织交界的浸润缘和慢性炎症组织,特别是纤维化组织.对照组正常胰腺组织除腺泡的微血管血供外,叶间结缔组织很少能观察到新生血管.p21ras阳性物可见于增生性胰腺导管上皮细胞及导管腺癌细胞,正常胰腺无表达,癌组织(47.2±9.4)和慢性胰腺炎组织(40.8±7.93)中微血管密度明显高于对照组(9.85±2.86)(P＜0.01).24例胰腺癌组织中p21ras阳性占15例,24例胰腺炎组织中p21ras阳性占11例,两者无显著差异,(x2=1.34,P＞0.05).p21ras阳性胰腺癌组织MVD(52.1±8.3)明显高于p21ras阴性的胰腺癌(39.1±4.1)及慢性胰腺炎组织(35.99±3.36)(P＜0.01),p21ras阳性的慢性胰腺炎组织MVD亦高于p21ras阴性的慢性胰腺炎组织(P＜0.01).结论微血管生成与K-ras突变关系密切,可能是胰腺癌发生发展中的早期事件,在慢性胰腺炎、胰腺癌的演变中亦起了不可低估的作用.     

Identification of immunoreactive pancreatic stone protein in pancreatic stone, pancreatic tissue, and pancreatic juice

We first examined whether pancreatic stone protein (PSP) was present in pancreatic stone and normal pancreatic tissue. By using HPLC and Western blotting, a protein of Mr 13.5 kDa that reacted with monoclonal antibody against PSP was detected as a major component in EDTA-soluble fractions of pancreatic stone. In an in vitro experiment, this protein dose-dependently suppressed CaCO3 precipitation. PSP was immunohistochemically stained in the acinar cells of normal pancreatic tissue. Based on these findings, it seemed that PSP in pancreatic stone is probably a physiological secretory protein of the pancreas. We subsequently examined immunoreactive PSP in normal pancreatic juice by the Western blotting method. In all of the specimens, the band for immunoreactive PSP in pancreatic juice was found to correspond to 13.5 kDa, which thus agreed with that of purified PSP from a stone.     

Interdigestive canine pancreatic juice composition and pancreatic reflux and pancreatic sphincter anatomy

Canine and human exocrine pancreatic secretion into the duodenum during fasting is cyclical and related to intestinal motility. To characterize the composition of pure pancreatic juice during the cyclically recurring sequence of propagated motor events (interdigestive motor complex) and to establish whether pancreatic reflux occurs, dogs were prepared with three permanent indwelling duodenal catheters and a pancreatodochal cutaneous catheter. The duodenal catheters were used to record duodenal pressures and measure pancreatic secretion of trypsin, lipase, and bicarbonate, based on the recovery of a constantly perfused marker, [¹⁴C]PEG. Pancreatic duct pressures or pancreatic juice concentrations of [¹⁴C]PEG, trypsin, lipase, or bicarbonate (done separately in each of five dogs throughout one interdigestive cycle on 4 different days) were related to duodenal motor activity. Finally, the pancreatic duct orifice of freshly sacrificed dogs was examined by light and electron microscopy. During fasting, (1) pancreatic volume secretion increased 10-fold during phases II, III, and IV (P<0.001), and bicarbonate concentration increased during phases III and IV (P<0.05) compared with phase I, while trypsin and lipase concentrations did not change; (2) reflux of duodenally perfused [¹⁴C]PEG into the pancreatic duct occurred in two of five dogs and was minimal (<0.1%); and (3) a positive mean pressure gradient from duodenum to pancreatic duct occurred only during phase III (7.4±4.1 cm H₂O). Anatomic studies of the pancreatic duct opening showed a specialized papillary mucosa and an independent crescentic sphincter muscle. We conclude that during fasting, pancreatic juice composition is intimately linked to the different phases of interdigestive intestinal motor activity and that an efficient antireflux mechanism exists.Supported in part by contract CP 55660 and grant CA 25064 from the National Institutes of Health, U.S. Public Health Service, Bethesda, Maryland.     

Serum pancreatic stone protein in pancreatic diseases

Summary Serum pancreatic stone protein (PSP) was determined in sera of pancreatic and nonpancreatic diseases using enzyme immunoassay specific to human PSP to study the diagnostic and pathophysiological significance of PSP. Serum PSP in acute pancreatitis (mean±SD=1075.4±2849.1 ng/mL,n=33) was significantly higher than that in controls (78.6±31.8 ng/mL,n=37,p<0.01), chronic pancreatitis (156.8±82.8 ng/mL,n=32,p<0.05), and pancreatic cancer (148.468.8 ng/mL,n=26,p<0.05). No significant difference was found between noncalcified and calcified chronic pancreatitis. Serum PSP levels were significantly higher in chronic renal failure under hemodialysis (1796.0±1492.9 ng/mL) than in other diseases such as peptic ulcer, liver cirrhosis, gallstone, and diabetes mellitus. Low but significant correlation was obtained between serum PSP and serum immunoreactive trypsin (r=0.22,p<0.05). Increased serum PSP levels in acute pancreatitis and chronic renal failure suggest that serum PSP levels reflect reflux from pancreatic secretion, release from damaged pancreatic acinar cells, or retention in circulation, and can be useful for diagnosis of acute pancreatitis, but not chronic calcified pancreatitis.     

Endoscopic pancreatic stenting in pancreatic cancer.

Most pancreatic carcinomas are unresectable at the time of diagnosis; therefore, palliative treatment is very often the main concern of clinicians in this setting. The main symptoms resulting in the need for palliation in pancreatic cancer are obstructive jaundice, duodenal obstruction and pain. Therapeutic endoscopy plays a major role in the palliation of obstructive jaundice by stent placement into the biliary ducts. Initial experience has also been gained recently with endoscopic placement of expandable metallic stents to treat gastric outlet obstruction. Much less is known about the possible role of endoscopic pancreatic stenting in patients with unresectable pancreatic carcinoma. The main indication for pancreatic ductal stenting is 'obstructive' pain related to meals in patients with dilated main pancreatic duct beyond the stricture and intraluminal brachyradiotherapy. The technique of endoscopic pancreatic stenting does not substantially differ from that applied on the biliary tree. When technically possible, placement of 10 French plastic stents is preferred. According to the authors' indications, only about 15% of patients with advanced pancreatic cancer (55 of 355 in the present study) may potentially benefit from this technique. Pancreatic stenting may be obtained in more than 80% of these selected patients, with low morbidity (less than 10%) and no procedure-related mortality. According to the authors of the present and other studies reported in the literature, about 60% of patients treated because of 'obstructive' pain become symptom-free, and another 20% to 25% significantly reduce the amount of analgesic drugs required. Intraluminal brachyradiotherapy with 192iridium in the main pancreatic duct is a feasible and safe method to deliver high radiation doses to the tumour while sparing adjacent organs. Brachyradiotherapy may be performed alone or in conjunction with external beam radiotherapy. Because of the small number of patients suitable for this treatment, only a multicentre study will be able to detect whether intraluminal brachyradiotherapy in pancreatic cancer may have any positive impact on survival.     

Solitary pancreatic tuberculosis mimicking advanced pancreatic carcinoma

A 40-year-old woman was referred for pancreatic head carcinoma invading the portal vein. The dichotomy between the radiological findings and the general condition of the patient, as well as the laboratory results (no evidence of cholestasis), cast doubt on the diagnosis. There was no history of tuberculosis. The chest radiograph revealed no pathological findings. The anatomic relationships of the lesion entailed a high risk of vascular injury if tissue biopsy were to be done; therefore, diagnostic laparotomy was performed. Biopsy revealed granulomas with caseous necrosis, consistent with tuberculosis. After 6 months of antituberculosis treatment, the lesions had completely resolved. Tuberculosis should be considered in the differential diagnosis of pancreatic masses, particularly in regions where the disease is endemic. The condition usually resembles an advanced pancreatic tumor. Performing a biopsy of inoperable lesions and maintaining a reasonable skepticism in regard to the evaluation of operable lesions (attention to nonexclusive but helpful clues, such as young patient age, history of tuberculosis, absence of jaundice) will lead to the diagnosis in most patients. Diagnostic laparotomy may be required in a small subset of patients. The response to antituberculosis treatment is very favorable. The role of resection (e.g., pancreatoduodenectomy) is very limited.