Identification of oligopeptide binding to colon cancer cells separated from patients using laser capture microdissection

The development of intravascular conjugates that efficiently deliver genes or drugs to tumors is limited by the lack of efficacious targeting ligands. Small targeting peptides, such as those iterated by phage display technology, offer enormous potential for these applications. The majority of reports published to date have focused on the identification of peptides isolated for their ability to bind to human cancer cell lines in vitro, and have failed to account for the loss of polarization and de-differentiation of such cells from their in vivo state. Here, we report a novel approach for the identification of peptides capable of binding specifically to cancer cells derived from clinically resected human colon cancer. In this strategy, laser capture microdissection (LCM) is performed on a surgically resected colon cancer specimen to separate only cancer cells from the specimen. Subsequently, biopanning was performed on the LCM-selected colon cancer cells to identify peptide sequences that bound specifically to them. A peptide containing the SPT motif was selected as the most promising consensus sequence binding specifically to the LCM-selected colon cancer cells. Phage clones displaying the SPT motif demonstrated 9-fold higher binding to colon cancer cells derived from a patient than insertless phage (p < 0.05), while, recovery of the SPT phage from the colon cancer cell lines DLD-1 and HCT-15 was 7-fold higher than that of the control insertless phage (p < 0.05). The binding of SPT phage to colon cancer cells from the patient was confirmed by immunofluorescence. Additionally, a synthesized SPT-containing peptide (SPTKSNS) showed binding activity in the absence of mitogenic effects on colon cancer cells in vitro. In summary, we have introduced LCM into a biopanning procedure and identified a small peptide that binds preferentially to colon cancer cells derived from a clinically resected sample. This procedure could be applicable for the design of customized cancer cell targeting methodologies using clinical biopsy samples from human subjects.     

开腹器械在36例腹腔镜胆总管切开取石术中的应用

目的探讨腹腔镜胆总管切开取石术(LCBDE)中应用开腹器械取石的可行性。方法回顾性分析我院36例LCBDE取石的临床资料。胆总管切开后以纤维胆道镜检查确定结石的部位、大小及数量,以常规开腹胆道取石器械进行胆道取石,并与同期36例开腹手术进行比较。结果LCBDE组36例手术均获成功,无中转开腹。常规于术后4周行T管造影,34例示结石取净;胆道残余结石2例,均于术后6周经T管窦道胆道镜取石治愈。在手术时间上,LCBDE组与开腹手术组相比无显著性差异(P>0.05);术后住院时间较开腹手术组明显缩短(P<0.05)。两组均留置T管,均无胆瘘、胆管狭窄或出血病例,均痊愈出院。LCBDE组36例随访6~48个月均无异常。结论严格选择手术适应证,应用开腹器械进行腹腔镜胆道取石安全有效、操作容易、成本低、手术时间短。     

腹腔镜下胆总管一期缝合在胆总管探查术中的应用

目的探讨腹腔镜胆总管探查术后一期缝合的可行性和临床价值。方法选取2014年1月~2017年6月46例因肝外胆管结石行腹腔镜胆总管探查后一期缝合的病例作回顾性分析。观察同期48例腹腔镜下探查+T管引流。对比观察两组术中情况(手术时间、出血量、取石成功率)、术后情况(胃肠功能恢复时间、术后并发症)和住院时间等指标。所有病例不含肝内胆管结石,术中经胆道镜或胆道造影排除胆道残石及狭窄,腔镜下一期缝合后小网膜孔常规放置多孔引流管。结果腹腔镜下一期缝合术后7例腹腔引流液含胆汁,均未特殊处理。术后平均住院日8.5d。所有病例术后门诊随访,3个月内复查B超和CT,未发现胆道残石,亦无一例发现肝外胆管狭窄。结论对经过严格选择的肝外胆管结石病例,经术中精细操作和术中胆道镜检查排除残石及下端狭窄后,腹腔镜下一期缝合可作为另一种理想的术式选择。     

腹腔镜胆总管切开取石胆总管一期缝合的临床研究

目的观察腹腔镜胆总管切开取石胆总管一期缝合的治疗效果。方法收集本院于2014年8月~2016年8月收治的50例胆总管结石患者。随机分为对照组和实验组,50例患者均采取腹腔镜下胆总管切开取石,对照组置入T管引流,实验组进行胆总管一期缝合,记录患者的手术时间、出血量、并发症及术后住院时间等情况,分析两组的临床疗效。结果 50例患者均在腹腔镜下顺利完成手术。比较两组的手术时间及术中出血量,差异无统计学意义(P>0.05)。对照组术后住院天数平均(9.28±1.90)d,实验组平均(6.04±1.06)d,两组比较差异有统计学意义(t=55.24,P<0.05)。对照组8例患者出现并发症,包括4例胆瘘和4例T管引流不畅,实验组并发症共2例,低于对照组,2例均为胆瘘,两组比较差异有统计学意义(χ~2=4.50,P<0.05)。均于术后随访半年到一年,行B超或EPCR检查,未发现胆总管结石残留。结论腹腔镜胆总管切开取石胆总管一期缝合术在胆总管结石治疗中的疗效显著,且安全可行,值得临床推广。     

Pharmacokinetics of phenazone (antipyrine) in rabbits with experimental common bile duct obstruction.

1. An altered functional state of liver due to experimental cholestasis could result in a change in the biotransformation of drugs. The aim of this study was to evaluate an influence of obstructive cholestasis on the pharmacokinetics of phenazone (antipyrine). 2. The investigation was carried out on male rabbits, randomly allocated into two groups: shamoperated and animals with biliary ducts ligation. Phenazone was administered intragastrically as a probe of drug metabolism. 3. Measurements, i.e. laboratory and pharmacodynamic tests, as well as pharmacokinetic assays, were performed before the operation as well as 10-12 days after the bile duct ligation. At the end of the study livers were examined macro- and microscopically and biochemical analysis of the liver microsomes was performed. 4. The measured pharmacokinetic parameters suggested an impaired biotransformation of phenazone in animals with obstructive cholestasis, leading to a slower drug elimination.     

Induction of bone loss by bile duct ligation in rats.

Metabolic bone loss is the most common complication of chronic liver disease. However, there is little information available about bone loss in rats with a ligated bile duct, a biliary-type of experimental cirrhosis model. Therefore, in this study, the effect of bile duct ligation (BDL) on bone mineral density (BMD) and plasma insulin-like growth factor-I (IGF-I) levels was examined in rats. Two weeks after BDL operation, the rats with a ligated bile duct showed a pronounced and significant decrease in both trabecular and cortical BMD of the femur. In these rats, decreases in food intake and plasma IGF-I levels plus elevated levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and bilirubin were also observed. When the effect of dietary restriction (30% or 50%) over 2 weeks was then investigated in normal rats, 50% food restriction resulted in a significant decrease in femoral trabecular bone density although BMD was unchanged by 30% dietary restriction. The plasma levels of IGF-I were also decreased in food-restricted rats. Thus, the decrease in femoral bone density in 50% food-restricted rats was less potent compared with BDL rats; nevertheless, the decrease in plasma levels of IGF-I was almost equally potent in both BDL and food-restricted rats. Overall, this study showed that BDL operation resulted in a pronounced bone loss in rats, but also that this bone loss might not be merely due to the decreases in food intake and plasma IGF-I levels. These results suggest that BDL in rats is a useful experimental cirrhosis model for evaluating the bone loss associated with chronic liver disease.     

手术治疗肝内胆管结石并胆管狭窄的效果分析

目的：评价手术治疗肝内胆管结石并胆管狭窄的临床效果。方法选取2011年3月∽2013年12月本院收治的肝内胆管结石并胆管狭窄患者128例,随机分为两组,各64例,对照组根据病情的不同,选择相应的手术方式;观察组在对照组的基础上进行术中胆道镜探查及取石处理。观察并比较两组的临床疗效、并发症发生率及复发情况。结果观察组的优良率为96.9%,明显高于对照组的84.4%,差异有统计学意义（P〈0.05）;观察组的并发症发生率低于对照组,两组比较差异有统计学意义（P〈0.05）;观察组的复发率为6.3%,对照组的复发率为14.1%,两组比较差异有统计学意义（P〈0.05）。结论采用合适的手术方式并在此基础上进行术中胆道镜探查及取石处理治疗肝内胆管结石并胆管狭窄的临床疗效好,并发症发生率及复发率低,值得临床推广应用。     

Detection of kidney damage by malathion impurities using a microdissection technique

O,O,S-Trimethylphosphorothioate (OOS-Me) and O,S,S-trimethylphosphorodithioate (OSS-Me) are impurities in technical grade malathion and related insecticides which have been shown to cause delayed death in rats following a single oral dose of 40-60 mg/kg. In connection with studies on the mode of action of these compounds, work on the microdissection and examination of nephrons was carried out. Nephrons from impurities-treated rats showed swelling, distortion and distension of glomeruli, as well as narrowing of the first part of the proximal tubule (swan neck). These results were similar to those observed from kidney tissue obtained from cadmium-chloride-treated rats and are indicative of OOS-Me and OSS-Me-induced kidney tubule damage.     

LC联合胆道镜探查术在胆囊结石合并胆总管结石中的应用

探讨腹腔镜胆囊切除术（LC）联合胆管镜探查术在胆囊结石合并胆总管结石中的应用效果。以我院2013年6月至2015年6月收治的90例胆囊结石合并胆总管结石患者为研究对象,随机分成观察组和对照组,各45例。对照组采用传统开腹手术,观察组采用腹腔镜胆囊切除术联合胆管镜探查术式,观察2组手术效果。观察组总有效率95.56%,明显优于对照组的82.22%（P＜0.05）;观察组并发症发生率2.22%,明显低于对照组的13.33%（P＜0.05）;观察组的手术时间、术后通气时间、术中出血量、住院时间比较均明显优于对照组（均P＜0.05）,2组住院费用差异不显著（P＞0.05）;虽然2组IgG、PCT指标均较治疗前升高,但观察组术后的升高幅度较对照组低（P＜0.05）。腹腔镜胆囊切除术联合胆管镜探查术治疗胆囊结石合并胆总管结石手术创伤小,术后恢复快,是一种有效的治疗方法。     

小切口胆囊切除术致胆道损伤的临床分析（附五例报告）

目的探讨小切口胆囊切除术致胆道损伤的原因及其预防措施和处理方法。方法选择948例行小切口胆囊切除术患者,发生胆道损伤5例,回顾性分析其临床资料。结果5例施行修补手术的患者,T管支撑时间为0．5～1a,拔管前胆道造影没有狭窄现象,拔管后无胆漏及梗阻性黄疸。5例患者均获得随访,随访时间为2～10a,无死亡,术后效果良好。结论小切口胆囊切除术虽具有切口小、恢复快、术后瘢痕小、费用低等优点,但是如果处理不当也会造成严重的胆道损伤。只要术者严格把握好手术适应征,仔细、规范操作,就可以减少胆道损伤;术后及时发现并妥善处理是减轻患者痛苦的关键。     

胆囊切除致肝外胆管损伤的外科处理

目的:探讨胆囊切除致肝外胆管损伤的处理方法.方法:对10例医源性胆管损伤的临床资料进行回顾性分析.结果:10例中8例为开腹胆囊切除术损伤,2例为腹腔镜胆囊切除术损伤.术中发现6例,术后早期确诊3例,后期确诊1例.手术方式:胆管端端吻合1例,胆总管十二指肠吻合1例,胆管壁修补1例,胆管空肠Roux-en-Y吻合7例.结论:正确掌握胆囊切除致肝外胆管损伤的处理原则及方法.胆管空肠Roux-en-Y是常用而有效的方法.     

胆囊切除术致胆管损伤的原因分析及处理

目的探讨胆囊切除术致胆管损伤的原因以及处理措施和时机。方法回顾性分析我院外科收治的24例胆管损伤患者的病因以及采取的再手术方式和时间,预后情况。结果11例胆囊切除术中发现胆管损伤的患者,立刻行胆管修复等手术,预后情况满意;13例术后发现胆管损伤的患者,采取胆管手术处理,只有1例因胆汁性肝硬化,胆管感染死亡,其他患者未出现胆管狭窄和胆管炎等并发症。结论在胆囊切除术中,要注意可能发生胆管损伤的危险因素,加以避免,如存在胆管损伤,力争早发现,早治疗,可有效的减少胆管狭窄,胆管炎等并发症,提高患者的生存质量。     

腹腔镜术中胆道镜技术联合钬激光碎石术治疗肝内胆管结石的可行性

目的探讨腹腔镜术中胆道镜技术联合钬激光碎石术治疗肝内胆管结石的可行性。方法选取2016年1月～2018年5月就诊于我院确诊为肝内胆管结石并拟行手术治疗的患者155例,随机分为研究组和对照组,对照组77例,研究组78例。两组患者均行腹腔镜术中胆道镜技术,在此基础上,对照组采用常规取石,研究组联合钬激光碎石术碎石,比较两组患者术中出血量、术后住院时间、腹腔引流管放置时间、疼痛评分、并发症发生率等指标。结果研究组患者术中出血量较对照组少,术后住院时间较对照组短,差异有统计学意义(P 0.05);研究组患者腹腔引流管放置时间短于对照组,疼痛评分低于对照组,差异有统计学意义(P 0.05);研究组患者术后并发症发生率显著低于对照组,差异有统计学意义(P 0.05)。结论肝内胆管结石患者行腹腔镜术中胆道镜技术联合钬激光碎石术治疗具有较高的临床治疗效果,可以有效减少术后并发症,促进术后恢复。     

胆道镜下钬激光碎石治疗难治性胆道残石体会

目的介绍纤维胆道镜下钬激光碎石治疗难治性胆道残石的经验体会。方法回顾分析3年来纤维胆道镜下钬激光碎石治疗难治性胆道残石临床资料。结果21例病例中胆残石取净者20例,胆残石未能取净者1例。全组无重大并发症。结论纤维胆道镜下钬激光碎石治疗难治性胆道残石不仅能降低结石残留,避免病人多次手术,而且具有安全性高、创伤小、费用低、术后恢复快等优点。     

双镜联合胆总管切开取石胆管一期缝合胆管自脱落支架置入术治疗胆总管结石的效果观察

目的探讨双镜联合胆总管切开取石胆管一期缝合胆管自脱落支架置入术治疗胆总管结石的临床效果。方法选取本院2011年1月~2014年8月收治的胆总管结石患者50例,其中20例（治疗组）行双镜联合胆总管切开取石胆管一期缝合胆管自脱落支架置入术,30例（对照组）行逆行胰胆管造影＋内镜下十二指肠乳头括约肌切开取石。比较两组的手术时间、住院时间、住院费用及并发症等。结果治疗组手术时间长于对照组,差异有统计学意义（P〈0.05）。两组住院时间、住院费用及并发症发生率差异无统计学意义（P〉0.05）。结论双镜联合胆总管切开取石胆管一期缝合胆管自脱落支架置入术具有手术成功率高、术后并发症少等优点,值得临床推广。     

超细胃镜在胆管疾病诊治中的应用效果

目的 探讨超细胃镜在胆管疾病诊治中的应用价值。方法 以2013年2月至2015年2月河南大学第一附属医院接受胃镜检查的60例胆管疾病患者为研究对象,随机双盲法将其分为十二指肠镜组（36例）与超细胃镜组（24例）,比较两组患者围检查期生命体征变化、反应分级,同时观察记录超细胃镜插镜情况及检查结果。结果 超细胃镜组24例一次性操作成功率100.0%,进镜视野A级占95.8%。3例超细胃镜直视下活检证实为腺瘤恶性病变,12例巨大嵌顿直视下激光碎石成功,3例疑似残余结石通过单纯胆总管探查排除结石残留。无胆管炎等并发症发生。超细胃镜组患者检查中收缩压为（96.40±9.18）mmHg,血氧饱和度为（97.85±1.37）%,优于十二指肠镜组的（103.42±10.74）mmHg、（95.71±3.05）%,差异均有统计学意义（P<0.05）;超细胃镜与十二指肠镜检查,患者反应分级差异无统计学意义（P>0.05）。结论 超细胃镜作为经口胆道镜诊治胆管疾病安全可行。     

Decompression of malignant biliary obstruction by duodenoscopic intubation of bile duct

Interest is increasing in non-operative methods of relieving malignant obstruction of the bile duct, and drainage tubes and prostheses may be placed in the bile duct via the percutaneous transhepatic route. Two cases are described, however, in which a duodenoscope was used and the approach was via the papilla of Vater. This method allows temporary or permanent drainage tubes to be placed through malignant strictures. This endoscopic approach is less invasive and should be safer than that by the transhepatic route; furthermore, removing and replacing a blocked endoprosthesis should be easier. Further study is needed, as the procedure is technically more difficult and its role in managing biliary strictures has yet to be defined.