老年病样本库规范化建设

目的探讨规范化建立老年病样本库、老年病生物样本库建立质量控制体系和样本信息化管理。方法制定老年病样本库标本库操作流程(SOP)及质量控制体系,规范化收集、处理及保存老年病患者的血清、血浆、核酸和组织等样本,应用信息管理软件进行样本信息录入。样本全程质控并随机抽样进行质量评估。结果自2013年建库以来,入库病种主要为少肌症、老年冠心病、老年糖尿病、肝癌、结肠癌、白塞氏病、帕金森病、阿尔茨海默病、脂肪肝等共16个病种。入库病例共2 110例,>60岁的体检人群约500例。入库样本共20 171份,并随机抽取30份DNA进行质量检测,结果表明所收集标本质量较好。建立了规范化的标本信息管理系统。结论通过老年病样本库规范化建立,能够建立有效的标准化的质量控制系统并规范化、有效地收集、保存和利用样本,为老年病研究提供宝贵资源和临床研究平台。     

肝癌生物样本库的标准化建立与信息化管理

目的 建立一个小型的拥有标准化操作流程和信息化管理功能的肝癌生物样本库。方法 根据纳排标准,在患者签署知情同意书基础上,收集2012年8月—2020年12月就诊于华中科技大学同济医学院附属同济医院肝脏外科的肝癌患者血液、组织和粪便样本,并按照全血、血清、冻存组织、石蜡包埋组织和粪便等不同样本的标准流程进行出入库管理,同时采集患者临床信息及其随访信息。通过随机抽取不同年份的肝癌及癌旁冷冻组织样本,检测总RNA的浓度和完整性,确保样本库冷冻样本保存质量。结果 101个自然月共收集4190例肝癌手术患者样本,其中包括41 718份冷冻组织样本、18 950份石蜡包埋组织样本、24 389份全血样本、20 060份血清样本和5392份粪便样本。所收集肝癌患者的年龄范围是13～88岁,男性占比85.1%,乙型肝炎和肝硬化的比例分别是83.3%和73.5%。根据样本的收集、处理、保存、应用及信息化管理,制定出一份标准的操作流程(SOP)和一套电子数据采集系统(EDC)。从样本库随机抽取的18份冷冻组织样本中,其中16份样本RNA质量较高,可满足后续实验需求。结论建立了标准化和信息化的肝癌生物样本库...     

Yes相关蛋白在结直肠癌组织中的表达及其临床意义

目的:研究结直肠癌变组织中Yes相关蛋白及mRNA的表达,探讨其临床意义.方法:收集2010-03/2011-03福建省南安市医院普外科及福建医科大学附属第一医院普外科80例结直肠癌患者术中切除的癌组织,同时收集20例癌旁组织和正常结直肠组织,应用免疫组织化学S-P法和原位杂交方法检测结直肠癌组织、癌旁组织和正常组织中Yes相关蛋白及mRNA的表达.原位杂交法和免疫组织化学S-P法均严格按照试剂盒说明操作.结果:结直肠癌组织中Yes相关蛋白及mRNA的阳性表达率分别为66.25%和68.75%,癌旁组织中Yes相关蛋白及mRNA的阳性表达率分别为20.0%和25.0%和正常结直肠黏膜组织Yes相关蛋白及mRNA的阳性表达率分别为10.0%和15.0%.结直肠癌组织中Yes相关蛋白及mRNA的阳性表达率明显高于癌旁组织和正常结直肠黏膜组织(P<0.01),癌旁组织与正常结直肠黏膜组织中Yes相关蛋白及mRNA阳性表达率比较差别无统计学意义.Yes相关蛋白及mRNA阳性表达率与年龄、性别、肿瘤部位之间比较差异无统计学意义,但与肿瘤分化程度和肿瘤分期等之间比较差异有统计学意义(P<0.05或0.01).结论:Yes...     

XAF1转录变异体在结直肠肿瘤中的差异表达

背景:X连锁凋亡抑制蛋白(XIAP)相关因子1(XAF1)能与XIAP结合而拮抗其caspase抑制活性,从而促进细胞凋亡,已被认定是一种肿瘤抑制基因。在多种肿瘤细胞中可检测到XAF1转录变异体,但不同转录变异体在结直肠肿瘤中的表达情况尚不明确。目的:检测XAF1及其转录变异体在不同结直肠组织中的表达,初步探讨其在结直肠肿瘤发生、发展中的作用。方法:收集结直肠癌及其配对癌旁组织、增生性息肉、腺瘤和正常结直肠黏膜组织样本,以免疫组化法和蛋白质印迹法检测XAF1蛋白表达,RT-PCR检测XAF1转录变异体表达。结果:与正常结直肠黏膜相比,XAF1蛋白在增生性息肉、腺瘤和癌组织中的胞核表达强度显著降低(P0.05),胞质表达强度有所增高(P0.05),总体表达强度显著降低(P0.05)。结直肠癌组织中的XAF1A蛋白表达显著低于相应癌旁组织(P0.05)。转录变异体XAF1A、XAF1B、XAF1C mRNA在结直肠肿瘤中的表达均显著低于正常结直肠黏膜(P0.05)。结论:XAF1及其转录变异体在结直肠肿瘤和正常结直肠黏膜中存在差异表达,腺瘤阶段即已出现XAF1表达下降并由胞核向胞质易位。转录后修饰可能影响XAF1基因功能。     

miR-574-3p在结直肠癌中的表达和作用机制研究

目的 检测并分析结直肠癌组织和细胞系中miR-574-3p的表达情况及其对结直肠癌细胞发生、发展的影响.方法 收集武汉市第三医院11对手术切除结直肠癌患者的癌组织和对应癌旁组织,3种结直肠癌细胞系和1种正常结直肠上皮细胞,通过实时荧光定量聚合酶链反应(qRT-PCR)检测临床样本及结直肠癌细胞系中miR-574-3p的...     

人结直肠癌微小核糖核酸的差异表达及其临床意义

目的:研究人结直肠癌、腺瘤和癌旁正常组织中微小核糖核酸(microRNAs,miRNAs)的差异表达谱,并初步探讨其临床意义.方法:选取2008-01/07苏州大学附属第一医院和泰州市人民医院结直肠癌、对应癌旁正常组织以及结直肠腺瘤标本,提取组织总RNA,采用illumina microRNA芯片技术检测3种不同类型组织中miRNAs的表达,采用实时定量PCR技术对芯片检测结果进行验证.将结直肠癌组织中异常表达的miRNAs与结直肠癌患者的临床病理资料进行分析.结果:3种不同类型结直肠组织中miRNAs表达有明显差异,结直肠癌与癌旁正常组织相比,有65个miRNAs表达异常(P<0.001),其中35个上调,30个下调,而腺瘤与正常结直肠组织相比,有55个miRNAs表达差异(P<0.001),其中上调29个,下调26个.有25个miRNAs相对于正常组织同时在结直肠癌和腺瘤中异常表达,其中高表达12个,低表达有13个.与腺瘤相比,结直肠癌中有25个miRNAs表达异常(P<0.01),其中13个上调,12个下调.进一步定量PCR验证结果显示与正常结直肠黏膜组织相比,在癌组织中miR-552,miR-142-...     

非酒精性脂肪性肝病生物样本库的建立与管理

目的探讨非酒精性脂肪性肝病(NAFLD)血液标本的采集和保存及生物样本库的建立和信息化管理方法。方法收集2009年10月-2013年10月经B超及临床血液学检测诊断为NAFLD的1226例患者的全血标本,进行生化指标测定、血浆及全血基因组DNA提取,并检测DNA的浓度和纯度,分别放至-80℃超低温冰箱储存备用,同时建立一套信息管理系统用于NAFLD生物样本库的管理。结果 1226例NAFLD患者的全血标本,其中包括双生子标本83例及家系标本100例。血浆及DNA的提取成功率均为100%,并随机抽取100份DNA进行检测,结果表明所收集标本能够满足后续试验要求。结论该研究成功建立了NAFLD生物样本库,拥有标准化的信息管理系统,对标本进行质量控制和信息化管理,为进一步研究NAFLD奠定了良好的基础。     

粪便DNA的Vimentin基因甲基化检测在结直肠癌患者早期诊断中的意义

目的通过检测结直肠癌组织及粪便中Vimentin基因甲基化探讨其在结直肠癌早期诊断中的应用。方法分别收集结直肠正常新鲜组织、癌新鲜组织及粪便标本,提取组织及粪便基因组DNA,DNA经亚硫酸盐处理后进行甲基化特异性PCR检测,检测结果经3%琼脂糖电泳鉴定。结果正常结直肠组织中未检测到Vimentin基因甲基化,结直肠癌组织中Vimentin基因甲基化的检出率为80%,结直肠癌患者粪便中Vimentin基因甲基化检出率为36%,其中早期直肠癌粪便的Vimentin基因甲基化检出率为66.7%。结论Vimentin基因在直肠癌早期诊断中具有重要意义。     

生物钟基因hClock蛋白在结直肠肿瘤中的表达研究

目的探讨人类生物钟基因hClock蛋白在结直肠肿瘤中的表达及其意义。方法应用免疫组织化学法检测不同Dukes分期结直肠肿瘤及相应癌旁组织中hClock基因蛋白产物（Clock蛋白）的表达，并比较其差异。结果Clock蛋白红结直肠肿瘤中呈现中或强阳性表达，与肿瘤分化程度及Dukes分期无显著相关性（P〉0.05）。癌旁组织呈现弱阳性表达。结论Clock蛋白与结直肠肿瘤的发生有相关性，与侵袭、转移的关系有待更深入地研究。     

Interplay between post-translational cyclooxygenase-2 modifications and the metabolic and proteomic profile in a colorectal cancer cohort

BACKGROUND Colorectal cancer(CRC) is the second most common cause of cancer death worldwide. It is broadly described that cyclooxygenase-2(COX-2) is mainly overexpressed in CRC but less is known regarding post-translational modifications of this enzyme that may regulate its activity, intracellular localization and stability. Since metabolic and proteomic profile analysis is essential for cancer prognosis and diagnosis, our hypothesis is that the analysis of correlations between these specific parameters and COX-2 state in tumors of a high number of CRC patients could be useful for the understanding of the basis of this cancer in humans.AIM To analyze COX-2 regulation in colorectal cancer and to perform a detailed analysis of their metabolic and proteomic profile.METHODS Biopsies from both healthy and pathological colorectal tissues were taken under informed consent from patients during standard colonoscopy procedure in the University Hospital of Bellvitge(Barcelona, Spain) and Germans Trias i Pujol University Hospital(Campus Can Ruti)(Barcelona, Spain). Western blot analysis was used to determine COX-2 levels. Deglycosylation assays were performed in both cells and tumor samples incubating each sample with peptide N-glycosidase F(PNGase F). Prostaglandin E2(PGE2) levels were determined using a specific ELISA. 1 H high resolution magic angle spinning(HRMAS) analysis was performed using a Bruker AVIII 500 MHz spectrometer and proteomic analysis was performed in a nano-liquid chromatography-tandem mass spectrometer(nano LC-MS/MS) using a QExactive HF orbitrap MS.RESULTS Our data show that COX-2 has a differential expression profile in tumor tissue of CRC patients vs the adjacent non-tumor area, which correspond to a glycosylated and less active state of the protein. This fact was associated to a lesser PGE2 production in tumors. These results were corroborated in vitro performing deglycosylation assays in HT29 cell line where COX-2 protein profile was modified after PNGase F incubation, showing higher PGE2 levels. Moreover,HRMAS analysis indicated that tumor tissue has altered metabolic features vs non-tumor counterparts, presenting increased levels of certain metabolites such as taurine and phosphocholine and lower levels of lactate. In proteomic experiments, we detected an enlarged number of proteins in tumors that are mainly implicated in basic biological functions like mitochondrial activity,DNA/RNA processing, vesicular trafficking, metabolism, cytoskeleton and splicing.CONCLUSION In our colorectal cancer cohort, tumor tissue presents a differential COX-2 expression pattern with lower enzymatic activity that can be related to an altered metabolic and proteomic profile.     

粪肠球菌预测结直肠癌复发的研究

目的 探索结直肠癌患者粪便样本中粪肠球菌丰度对于患者复发的预测意义.方法 选取哈尔滨医科大学附属第四医院普外科结直肠癌复发患者与非复发患者的粪便样本共103例,通过16 s测序技术寻找差异的肠道菌群,通过统计学分析探索差异菌群与患者临床病理学指标及无复发生存之间的关系.结果 粪肠球菌在复发患者粪便组织中丰度更高;粪便样...     

Chymase蛋白在结直肠癌组织中的表达及意义

目的探讨Chymase蛋白在结直肠癌组织中的表达及临床意义。 方法选取2010年9月至2012年9月于中国医学科学院肿瘤医院结直肠外科和哈尔滨医科大学附属第二医院结直肠肿瘤外科接受结直肠癌根治术的135例结直肠癌患者,通过免疫组化的方法检测Chymase蛋白在结直肠癌组织中的表达,分析Chymase蛋白与临床病理资料及预后的关系。 结果Chymase蛋白在结直肠癌组织中的表达低于癌旁组织,表达差异具有统计学意义（Χ²=4.305,P=0.038）,结直肠癌组织中,有远处转移患者Chymase蛋白表达降低（P=0.002）,Chymase蛋白的表达与N分期有关,分期越晚,Chymase蛋白的表达越低（Χ²=54.81,P<0.001）,在Ⅲ＋Ⅳ期患者Chymase蛋白表达降低（Χ²=50.84,P<0.001）,生存分析结果提示Chymase蛋白低表达患者预后不佳（Χ²=10.501,P=0.001）。 结论Chymase蛋白可能成为结直肠癌患者预后的分子靶标。     

长链非编码RNA在结直肠癌中的研究现状

近年来研究发现,长链非编码RNA在多种恶性肿瘤的发生发展中发挥重要作用,有望成为新的肿瘤治疗靶点,已成为目前肿瘤研究的热点问题之一。本文对长链非编码RNA的概况及其在结直肠癌预后判断、恶性潜能及诊断中的应用现状做一综述,为结直肠癌的精准治疗提供新的思路与依据。     

Down-regulation of DOC-2 in colorectal cancer points to its role as a tumor suppressor in this malignancy

PURPOSE: Colorectal cancer is the second most common cause of cancer death and the fourth most prevalent carcinoma in the Western world. Loss of tumor-suppressor gene function in colon cancer leads to ineffective negative growth regulation that is normally exerted by growth-suppressing factors. DOC-2/hDAB2 is a newly identified candidate tumor-suppressor gene in ovarian cancer and choriocarcinoma. In these tumors it negatively influences mitogenic signal transduction of growth factors and blocks ras activity. In the present study we sought to determine the role of DOC-2 in colorectal cancer. METHODS: DOC-2 expression was analyzed by Northern blot analysis, hybridization, and immunohistochemistry in 27 primary and metastatic colorectal cancers and in 15 normal colon tissues in correlation with clinicopathologic data. RESULTS: Northern blot analysis demonstrated a decrease of DOC-2 messenger RNA levels in primary and metastatic colorectal cancers compared with normal controls. In normal colorectal tissues, DOC-2 immunoreactivity was strongly present on the surface columnar epithelial cells. In contrast, DOC-2 immunoreactivity was weak to moderate in the epithelium of colorectal cancers, and the intensity of the signals in colorectal cancer was greatly decreased compared with the normal colorectal tissues. In addition, DOC-2 immunoreactivity in lymph node and liver metastasis was weak to absent in the cancer cells and significantly decreased compared with their primary tumors. CONCLUSIONS: The expression of DOC-2 is down-regulated in primary tumors and metastases of colorectal cancer, which suggests that DOC-2 functions as a tumor suppressor in this malignancy.     

Undetectable expression of hMLH1 protein in sporadic colorectal cancer with replication error phenotype

PURPOSE: Four DNA mismatch repair genes have been identified as being susceptible genes for hereditary nonpolyposis colorectal cancer. Deficiency of one of the mismatch repair genes causes the replication error phenotype in more than 80 percent of patients with hereditary non-polyposis colorectal cancer and in 10 to 30 percent of patients with sporadic colorectal cancer. To determine which mismatch repair gene is lacking the function in patients with replication error-positive colorectal cancer, several approaches have been used at the nucleic acid and protein levels. We studied replication error in 40 samples of randomly selected colorectal cancers and expression of hMSH2 and hMLH1 proteins analyzed by immunoblot in the tumor and normal tissues of the replication error-positive and replication error-negative samples. MATERIALS AND METHODS: Frozen tumor and normal tissues were obtained from 40 Japanese patients who had colorectal cancer. According to the Amsterdam criteria, those patients were classified as having 39 sporadic and 1 unknown colorectal cancers. Genomic DNA was extracted from tumor and normal tissues for determining replication error with eight microsatellite markers. Expression of hMSH2 and hMLH1 proteins in cell lysates of tumor and normal tissues of 16 patients was analyzed by immunoblot. RESULTS: The replication error phenotype was found in 6 (15 percent) of the 39 sporadic cases. hMLH1 protein was not detected in two of the six replication error-positive tumor tissues and not in the normal tissues, indicating that the tumor cells of the two patients had severe mutations in both alleles of thehMLH1gene. Another four replication error-positive and ten replication error-negative tumors and normal tissues expressed hMLH1 protein. hMSH2 protein was detected in all samples. CONCLUSION: hMLH1 protein was undetectable in the two tumor tissues of the six replication error-positive samples of sporadic colorectal cancer. The detection procedure used here may have potential use for determining a dysfunctional mismatch repair gene product.     

手助腹腔镜与全腹腔镜外科治疗结直肠癌的对比研究

目的比较手助腹腔镜与全腹腔镜在外科治疗结直肠癌中的近期疗效,另外对手辅助腹腔镜外科治疗结直肠癌的安全性做出评估。 方法选取2011年11月至2014年4月哈尔滨医科大学附属第二医院结直肠肿瘤外科的79例结直肠癌患者,其中接受腹腔镜下结直肠癌根治术者41例,接受手辅助腹腔镜下结直肠癌根治术者38例。根据2种术式分为全腹腔镜组（LAC组）和手助腹腔镜组（HALS组）。通过回顾性分析,比较2组患者的一般资料包括年龄、性别、肿瘤位置、肿瘤病理类型、分期、ASA分级、BMI指数等;手术资料包括手术时间、麻醉时间、术中出血量、中转开腹率、取标本切口长度、Trocar数目等;术后资料包括术后第一次排气时间、术后住院时间、住院费用、术后并发症情况等。 结果HALS组和LAC组两组手术时间、麻醉时间、术中出血量、Trocar使用数目、取标本切口长度、术后首次排气时间存在差异（P<0.05）;术后住院时间、住院费用、术后并发症差异无统计学意义（P>0.05）。 结论手辅助腹腔镜下结直肠癌根治术与全腹腔镜下结直肠癌根治术相比具有手术时间短、创伤小、术后恢复快等优点,是一种安全有效,可靠的技术。     

Building a bioresource for esophageal research: lessons from the early experience of an academic medical center

The establishment of biorepositories, linked to clinical and epidemiologic data, are central to the goals of personalized medicine and individualized cancer therapy. Repositories of DNA, RNA, and serum samples are valuable resources for cancer research, enabling the investigation of the underlying causes of cancer development, progression, and prognosis, as well as providing a resource for the investigation of biomarkers for early detection and prediction of response. With a greater reliance on sample‐derived data for molecular‐based research and clinical care, improved standards and informatics for sample procurement, storage, and analysis are necessary to maximize the value of tissue collection for research participants, investigators, and academic medical centers. We present herein the experience of an academic medical center in establishing a repository for esophageal research, with discussion of elements to be considered when establishing such a resource, from the quality assurance of samples to the organized collection and storage of associated clinical data. The development of this biorepository required significant planning to identify and consent participants by dedicated clinical and research personnel. Ensuring the quality of any biobank is of utmost importance, and one must understand the sample variability that exists during the acquisition of biospecimens. The time and type of fixative have been optimized in our unit by standard operating protocols. Methods for biomolecule extraction were tested by examining both the quality and the quantity of recovered sample. These procedures were overseen by a designated biobank manager, responsible for the acquisition of the sample from surgery, which limits variability in sample collection. Our unit also has a dedicated database manager for the maintenance of quality clinical data linked to the bioresource. The development and expansion of such repositories, at local and national levels, is required to enable leading academic medical centers and their investigators to provide optimal and molecularly guided care to their patients.