Detection of k-ras gene point mutation in fine needle aspiration and pancreatic juice by sequence special primer method and its clinical significance

INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected the presence of k-ras gene

Study on chronic pancreatitis and pancreatic cancer using MRS and pancreatic juice samples

AIM:To investigate the markers of pancreatic diseases and provide basic data and experimental methods for the diagnosis of pancreatic diseases.METHODS:There were 15 patients in the present study,among whom 10 had pancreatic cancer and 5,chronic pancreatitis.In all patients,pancreatic cancer or chronic pancreatitis was located on the head of the p-a-ncrea-s.Pa-thology da-ta-of a-ll p-a-tients wa-s confirmed by biopsy and surgery.Among the 10 patients with pancreatic cancer,3 people had a medical history of longterm alcohol consumption.Of 5 patients with chronic pancreatitis,4 men suffered from alcoholic chronic pancreatitis.Pancreatic juice samples were obtained from patients by endoscopic retrograde cholangiopancreatography.Magnetic resonance spectroscopyn was performed on an 11.7-T scanner (Bruker DRX-500) using Call-Purcell-Meiboom-Gill pulse sequences.The parameters were as follows:spectral width,15 KHz;time domain,64 K;number of scans,512;and acquisition time,2.128 s.RESULTS:The main component of pancreatic juice included leucine,iso-leucine,valine,lactate,alanine,acetate,aspartate,lysine,glycine,threonine,tyrosine,histidine,tryptophan,and phenylalanine.On performing 1D 1H and 2D total correlation spectroscopy,we found a triplet peak at the chemical shift of 1.19 ppm,which only appeared in the spectra of pancreatic juice obtained from patients with alcoholic chronic pancreatitis.This triplet peak was considered the resonance of the methyl of ethoxy group,which may be associated with the metabolism of alcohol in the pancreas.CONCLUSION:The triplet peak,at the chemical shift of 1.19 ppm is likely to be the characteristic metabolite of alcoholic chronic pancreatitis.

Potential prognostic biomarkers in pancreatic juice of resectable pancreatic ductal adenocarcinoma

Despite potentially curative surgery pancreatic cancer has a dismal prognosis.Serum cancer antigen 19-9(CA 19-9) correlates with tumor burden,resectability and survival in patients with pancreatic ductal adenocarcinoma.Identification of novel biomarkers may facilitate early diagnosis of pancreatic cancer and improve survival.Pancreatic juice is a rich source of cancer-specific proteins rendering it a promising tool for identifying biomarkers.Recent proteomic and micro RNA expression analyses have identified several biomarkers of potential diagnostic and prognostic value.Tumor markers CA 19-9 and carcinoembryonic antigen(CEA) are widely used in the characterization of premalignant and malignant lesions of the pancreas.Elevated level of CEA in bile is a marker for malignancy and a predictor of hepatic recurrence.The potential value of CA 19-9,CEA and lactate dehydrogenase as prognostic biomarkers in pancreatic juice and bile is unknown.Specimens of pancreatic juice and bile can be readily collected during surgical resection of the tumor.Profiling of pancreatic juice and bile to identify novel prognostic biomarkers may improve selection of patients for adjuvant therapy with a favorable impact on overall survival in patients diagnosed with pancreatic cancer.

Impact of continuous local lavage on pancreatic juice-related postoperative complications: Three case reports

BACKGROUND Postoperative pancreatic leakage readily results in intractable pancreatic fistula and subsequent intraperitoneal abscess.This refractory complication can be fatal;therefore,intensive treatment is important.Continuous local lavage (CLL) has recently been reevaluated as effective treatment for severe infected pancreatitis,and we report three patients with postoperative intractable pancreatic fistula successfully treated by CLL.We also discuss our institutional protocol for CLL for postoperative pancreatic fistula.CASE SUMMARY The first patient underwent subtotal stomach-preserving pancreaticoduodenectomy,and pancreatic leakage was observed postoperatively.Intractable pancreatic fistula led to intraperitoneal abscess,and CLL near the pancreaticojejunostomy site was instituted from postoperative day (POD) 8.The abscess resolved after 7 d of CLL.The second patient underwent distal pancreatectomy.Pancreatic leakage was observed,and intractable pancreatic fistula led to intraperitoneal abscess near the pancreatic stump.CLL was instituted from POD 9,and the abscess resolved after 4 d of CLL.The third patient underwent aneurysmectomy and splenectomy with wide exposure of the pancreatic parenchyma.Endoscopic retrograde pancreatic drainage was performed on POD 15 to treat pancreatic fistula;however,intraperitoneal abscess was detected on POD 59.We performed CLL endoscopically via the transgastric route because the percutaneous approach was difficult.CLL was instituted from POD 63,and the abscess resolved after 1 wk of CLL.CONCLUSION CLL has therapeutic potential for postoperative pancreatic fistula.

Early detection of pancreatic cancer

The diagnosis of pancreatic cancer associates an appalling significance.Detection of preinvasive stage of pancreatic cancer will ameliorate the survival of this deadly disease.Premalignant lesions such as Intraductal Papillary Mucinous Neoplasms or Mucinous Cystic Neoplasms of the pancreas are detectable on imaging exams and this permits their management prior their invasive development.Pancreatic intraepithelial neoplasms(PanIN)are the most frequent precursors of pancreatic adenocarcinoma(PDAC),and its particular type PanIN high-grade represents the malignant non-invasive form of PDAC.Unfortunately,PanINs are not detectable on radiologic exams.Nevertheless,they can associate indirect imaging signs which would rise the diagnostic suspicion.When this suspicion is established,the patient will be enrolled in a follow-up strategy that includes performing of blood test and serial imaging test such as computed tomography or magnetic resonance imaging,which will cost in the best-case scenario a burden of healthcare systems,and potential mortality in the worst-case scenario when the patient underwent resection surgery,worthless when there is no moderate or high grade dysplasia in the final histopathology.This issue will be avoid having at its disposal a diagnostic technique capable of detecting high-grade PanIN lesions,such is the cytology of pancreatic juice obtained by nasopancreatic intubation.Herein,we review the possibility of detection of early malignant lesions before they become invasive PADC.

Role of pancreatic juice cytology in diagnosis of high-grade pancreatic intraepithelial neoplasia

High-grade pancreatic intraepithelial neoplasia is a challenging diagnosis and itdoes not exhibit mass lesions. It is suspected based on changes in the mainpancreatic duct in magnetic resonance cholangiopancreatography. Sometimesonly an unclear duct shows in magnetic resonance cholangiopancreatographywith no focal strictures and upstream dilatation of the main pancreatic duct. Serialpancreatic juice cytology is valuable in diagnosis of those patients.

Role of the preoperative usefulness of the pathological diagnosis of pancreatic diseases

Pancreatic cancer is the fifth leading cause of cancer death and has the lowest survival rate of any solid cancer.Endoscopic ultrasound-guided fine-needle aspiration biopsy(EUS-FNA) is currently capable of providing a cytopathological diagnosis of pancreatic malignancies with a higher diagnostic power,with a sensitivity and specificity of 85%-89% and 98%-99%,compared to pancreatic juice cytology(PJC),whose sensitivity and specificity are only 33.3%-93% and 83.3%-100%.However,EUS-FNA is not effective in the cases of carcinoma in situ and minimally invasive carcinoma because both are undetectable by endoscopic ultrasonography,although PJC is able to detect them.As for the frequency of complications such as post endoscopic retrograde cholangiopancreatography pancreatitis,EUS-FNA is safer than PJC.To diagnose pancreatic cancer appropriately,it is necessary for us to master both procedures so that we can select the best methods of sampling tissues while considering the patient's safety and condition.

Evaluation of diagnostic cytology via endoscopic naso-pancreatic drainage for pancreatic tumor

AIM: To evaluate the usefulness of cytology of the pancreatic juice obtained via the endoscopic naso-pancreatic drainage tube(ENPD-C).METHODS: ENPD was performed in cases where a diagnosis could not be made other than by using en-doscopic retrograde cholangiopancreatography and in cases of pancreatic neoplasms or cystic tumors, includ-ing intraductal papillary mucinous neoplasm(IPMN) suspected to have malignant potential. 35 patients(21 males and 14 females) underwent ENPD between January 2007 and June 2013. The pancreatic duct was imaged and the procedure continued in one of ENPD-C or ENPD-C plus brush cytology(ENPD-BC). We checked the cytology result and the final diagnosis.RESULTS: The mean patient age was 69 years(range, 48-86 years). ENPD-C was performed in 24 cases andENPD-C plus brush cytology(ENPD-BC) in 11 cases. The ENPD tube was inserted for an average of 3.5 d. The final diagnosis was confirmed on the basis of the resected specimen in 18 cases and of follow-up findings at least 6 mo after ENPD in the 18 inoperable cases. Malignancy was diagnosed in 21 cases and 14 patients were diagnosed as having a benign condition. The ratios of class Ⅴ/Ⅳ:Ⅲ:Ⅱ/Ⅰ?findings were 7:7:7 in malignant cases and 0:3:11 in benign cases. The sensitivity and specificity for all patients were 33.3% and 100%, re-spectively. The cytology-positive rate was 37.5%(6/16) for pancreatic cancer. For IPMN cases, the sensitivity and specificity were 33% and 100%, respectively.CONCLUSION: Sensitivity may be further increased by adding brush cytology. Although we can diagnosis cancer in cases of a positive result, the accuracy of ENPD-C remains unsatisfactory.

Preventing pancreatic fistula after distal pancreatectomy: An invagination method

Following an increase in the use of the GIA stapler for treating a pancreatic stump, more techniques to prevent postoperative pancreatic juice leakage have been required. We describe one successful case using our new technique of invaginating the cut end of the pancreas into the stomach to prevent a pancrea-tic fistula(PF) from occurring. A 50-year-old woman with pancreatic cancer in the tail of the pancreas underwent distal pancreatectomy, causing a grade A PF. We resected the distal pancreas without additional reinforcement to invaginate the stump into the gastric posterior wall with single layer anastomosis using a 3-0 absorbable suture. The drain tubes were removed on the third postoperative day. Although a grade A PF was noted, the patient was discharged on foot on the eleventh postoperative day. Our technique may be a suitable method for patients with a pancreatic body and tail tumor.

The Influence of Feed Intake and Conditioned Reflex on Exocrine Pancreatic Function in Broiler Chicks

The data of an experimental study of the influence of feeding and the conditional stimulus on digestive function in birds is presented. It is stated that feed consumption is a powerful stimulator of pancreatic secretion in broiler chicks. The secretory response of the pancreas was the highest in the morning when secretion of pancreatic juice was 1.8 times higher, amylase activity 3.2 times, proteases activity 3.3 times, lipase activity 2.1 times higher than before feeding (basal levels). The regulation of pancreatic activity is known to be complex-reflex with conditioned-reflex component being very important since it provides 25% higher juice secretion and 42% - 74% higher enzyme secretion compared to the basal levels during few minutes after the conditional signal.

Detecting K-ras and p53 gene mutation from stool and pancreatic juice for diagnosis of early pancreatic cancer

OBJECTIVE: To explore new methods for the early diagnosis of pancreatic cancer through detection of K-ras and p53 mutations in pancreatic juice and stool. METHODS: 201 patients in PUMC Hospital from 1994 - 2000 and 60 control individuals were enrolled in this study. K-ras point mutation was detected by PCR-RFLP while p53 mutation was detected by PCR-SSCP. RESULTS: K-ras mutation was found in pancreatic juice in 87.8% (36/41) of pancreatic cancer patients and 23.5% (4/17) of benign pancreatic disease patients. In 261 stool specimens, amplification found mutations successfully in 235 patients (90%). K-ras mutation was found in stool in 88% (66/75) of pancreatic cancer patients, 51.1% (24/47) of benign pancreatic disease patients and 19.6% (9/46) of normal individuals. p53 mutation was found in pancreatic juice in 47.4% (18/38) of pancreatic cancer patients and 12.5% (2/16) of benign pancreatic disease patients. p53 mutation was found in stool in 37.1% (23/62) and 19.1% (4/21) of chronic pancreatitis patients. CONCLUSION: K-ras mutation in pancreatic juice has higher diagnosis sensitivity and specificity, and therefore may be used as a supplement in the diagnosis of pancreatic cancer. Detection of K-ras mutation combined with p53 mutation in stool can aid in the screening of pancreatic cancer.

Pharmacokinetics of 5-fluorouracill and its penetration into pancreatic juice in dogs

研究胰液中５氟脲嘧啶（５ＦＵ）的药物动力学行为及其与血药浓度的关系，并评价其穿透胰液特性．方法：杂种狗８只，手术放置胰外引流管后单剂量静注５ＦＵ２５０ｍｇ．采集血和胰液并用ＨＰＬＣ法测定药物浓度．计算药物动力学参数并作统计学分析．结果：血浆和胰液中５ＦＵ的平均Ｋ１０分别为９４ｈ－１和１０２ｈ－１（Ｐ＞００５）．血中和胰液药物消除均符合非线性模型，穿透指数为３３９±２８４．５ＦＵ由血浆向胰液穿透较迅速，胰液中的药物浓度持续高于同期血药浓度．结论：胰液中药物消除与血液相似，二者为同一动力学房室．

Stress kinase inhibition modulates acute experimental pancreatitis

AIM: To examine the role of p38 during acute experimental cerulein pancreatitis. METHODS: Rats were treated with cerulein with or without a specific JNK inhibitor (CEP1347) and/or a specific p38 inhibitor (SB203580) and pancreatic stress kinase activity was determined. Parameters to assess pancreatitis included trypsin, amylase, lipase, pancreatic weight and histology. RESULTS: JNK inhibition with CEP1347 ameliorated pancreatitis, reducing pancreatic edema. In contrast, p38 inhibition with SB203580 aggravated pancreatitis with higher trypsin levels and, with induction of acinar necrosis not normally found after cerulein hyperstimulation. Simultaneous treatment with both CEP1347 and SB203580 mutually abolished the effects of either compound on cerulein pancreatitis. CONCLUSION: Stress kinases modulate pancreatitis differentially. JNK seems to promote pancreatitis development, possibly by supporting inflammatory reactions such as edema formation while its inhibition ameliorates pancreatitis. In contrast, p38 may help reduce organ destruction while inhibition of p38 during induction of cerulein pancreatitis leads to the occurrence of acinar necrosis.

胰液分子生物标志物在诊断胰腺癌中的研究进展

胰腺癌(pancreatic cancer,PC)是全世界最致命的恶性肿瘤之一,因其起病隐匿,恶性程度高,致死率高,被称为“癌中之王”。胰腺癌恶性程度高,进展迅速,但起病隐匿,早期症状不典型,常规影像学检查难以发现早期病变,通常只有出现黄疸以及腹部、背部疼痛等症状就诊时胰腺癌才得以发现。手术切除是目前胰腺癌唯一的治愈方案,但由于早期诊断困难,绝大多数患者在确诊时已属于中晚期,失去了手术治愈的机会。研究证实胰腺癌的进展是相对比较缓慢的,其起始的肿瘤细胞至少需要15年才能获得转移能力。因此,如果能够通过肿瘤标志物使胰腺癌得以及时发现,就能够有效提高胰腺癌患者的生存期。目前临床上应用最普遍且诊断价值较高的肿瘤标志物为糖类抗原199 (Carbohydrate antigen199,CA19-9),因约3%~7%的胰腺癌患者为Lewis抗原阴性血型结构而不表达CA19-9等原因,其敏感度仅79%~81%,不能为胰腺癌的诊断提供良好的效能。胰液作为一种肿瘤附近的体液,因其是肿瘤相关生物标志物的良好来源,引起了大量的关注。许多研究证实了胰液中的蛋白质、DNA以及外泌体等物质用以诊断胰腺癌所具有的良好准确性,胰液具有成为胰腺癌诊断的肿瘤标志物来源的良好前景。该文对胰液作为标本诊断胰腺癌的临床价值进行综述。

胰液K-ras12密码子点突变检测对胰腺癌诊断的临床价值

目的 :探讨胰液中 K- ras 12密码子点突变对胰腺癌诊断的临床应用价值。 方法 :采用内镜下置鼻胰管方法收集 2 0例胰腺疾病的胰液标本 ,聚合酶链反应 -限制性片段长度多态性分析 (PCR- RFL P)检测胰液 K- ras基因第 12密码子点突变 ,与肿瘤标记物 CA 19- 9及 CEA检测结果比较。结果 :12例胰腺癌患者胰液标本中 K- ras突变率为 5 8.3% (7/ 12 ) ;8例慢性胰腺炎患者标本 K- ras突变率为 12 .5 % (1/ 8) ,胰腺癌胰液 K- ras基因突变检测的敏感性为 5 8.3% ,特异性为 87.5 % ,阳性预示值为 87.5 % ,阴性预示值为 5 8.3%。同一组病例以血清 CA19- 9>37U / m l、CEA >4 .5μg/ ml为阳性界值。胰腺癌血清CA19- 9、CEA阳性率分别为 5 8.3% (7/ 12 )、4 1.7% (5 / 12 )。结论 :K- ras基因突变与胰腺癌相关性好 ,胰液中 K- ras 12密码子突变率高 ,特异性强 ,为胰腺癌早期诊断的初步筛选提供了手段。

二胺氧化酶在急性坏死性胰腺炎肠道损伤中的作用

目的探讨急性坏死性胰腺炎（ＡＮＰ）时血浆二胺氧化酶（ＤＡＯ）水平变化及其与肠道损伤的关系．方法通过向主胰管内注入牛磺胆酸钠和胰蛋白酶复制犬ＡＮＰ模型（ｎ＝８）；检测血清淀粉酶水平；测定血浆、肠组织ＤＡＯ活性；测量回肠粘膜绒毛形态学改变．结果ＡＮＰ发病后ｄ１（ｍｏｌ／ｓ，００２５±０００６ｖｓ００１１±０００２），ｄ２（ｍｏｌ／ｓ，００１９±０００５ｖｓ００１２±０００３）血浆ＤＡＯ活性增加，ｄ４（ｍｏｌ／ｓ，０００６±０００１ｖｓ００１４±０００２）以后明显下降，肠组织ＤＡＯ活性亦显著下降；回肠粘膜绒毛萎缩、脱落，高度、宽度降低，面积减少；血清淀粉酶水平升高３～１０倍；肝、胰、脾、肺、肾及肠系膜淋巴结出现细菌移位．结论ＡＮＰ时血中ＤＡＯ活性变化可反映肠粘膜上皮损伤。

胰液中端粒酶及亚单位检测对胰腺癌的诊断价值

目的 探讨检测胰液中端粒酶逆转录酶 (hTERT)mRNA表达和端粒酶活性对胰腺癌的诊断与鉴别诊断价值。方法 对比分析 2 4例胰腺癌和 14例慢性胰腺炎患者胰液中的hTERTmRNA和端粒酶活性的检测结果。结果 hTERTmRNA在胰腺癌中表达阳性率为 87.5 % ( 2 1/ 2 4) ;慢性胰腺炎中阳性率为 2 1.4% ( 3 / 14 ) (P <0 .0 0 1)。胰腺癌与慢性胰腺炎胰液中端粒酶阳性检出率分别为83 .3 % ( 2 0 / 2 4) ,2 8.6% ( 4 / 14 ) (P <0 .0 0 1)。结论 胰液中检测hTERTmRNA表达和端粒酶活性对胰腺癌诊断与鉴别诊断有一定价值。

鹅胰液的分泌及其调节研究

选成年杂交 (川白×太湖 )鹅 8只 ,用胰腺—十二指肠长久性回流瘘管手术 ,在不限制鹅运动和采食情况下收集纯净的胰液。实验期饲以大麦 (占 45 % )基础日粮 ,白天 (8:0 0～ 2 0 :0 0 )给食 ,夜间 (2 0 :0 0～ 8:0 0 )禁食 ,自由饮水。实验期连续收集胰液。结果表明 :鹅的胰液呈不规则的连续性分泌。其分泌速率白天为 (30 7 5 9± 2 9 40 ) μL/h ,夜间为 (10 0 0 0± 2 2 45 ) μL/h ;分泌量白天占 73 6 6 % ,夜间占 2 6 34%。皮下注射硫酸阿托品 0 5mg/只 ,则分泌速率分别降至对照期的 2 4 0 0 % (白天 )和 2 8 77% (夜间 ) (P <0 0 1) ;胰液蛋白酶、脂肪酶和淀粉酶的活力也较对照期下降(P <0 0 1)。表明鹅的胰液分泌受胆碱能神经调节。口服丙谷胺 15 0mg/kg ,胰液分泌速率试验期与对照期差异不显著(P >0 0 5 ) ;而胰液蛋白酶、脂肪酶和淀粉酶的活力显著下降 (P <0 0 1)。表明丙谷胺通过阻断CCK受体而影响胰酶的分泌 。

大山楂丸对消化系统的药理作用

本文以大山楂丸和其煎剂为主,观察了它们的消化系统的药理作用:①体外实验表明,丸剂和煎剂均能增强胃蛋白酶活性;②煎剂能增强胰脂肪酶活性;③丸剂、煎剂均能促进小鼠胃肠蠕动;④煎剂口服给药并不增加正常大鼠胃液分泌量,对胃液中的游离酸、总酸及胃蛋白酶含量均无明显影响。本文还比较了该方丸剂、煎剂与单味提取配方液的药理作用。

口服果汁后MRCP评价胰管形态学的临床研究

目的 评价口服果汁后MRCP对胰管成像的价值。 资料与方法 31例受试者口服果汁前后分别行MR-CP检查,评价胰管各部分显示情况和直径,胰管最佳显示时间,以及胃肠道重叠干扰情况。 结果 所有受试者口服果汁后胰管各部分显示情况及平均直径均比口服前显著提高,口服后胰管最佳显示时间为(55.75±15.65) min,胃肠道内液体重叠干扰最明显的时间是(43.88±17.12) min。 结论 口服果汁后MRCP可显著提高胰管显示情况,胰管显示最佳时间为口服后60 min。