门静脉高压症标本库的初步建立、质控及管理

目的 探索建立门静脉高压症(portal hypertension,PH)生物标本库及数据信息化管理方法和标准化操作流程,为PH基础与临床研究提供高质量的临床标本资源.方法 收集PH患者的血液、尿液、唾液、组织标本,将处理好的标本于-80℃冰箱保存,新鲜组织标本置于液氮保存,定期随机抽取标本进行质量控制.结果 现收集并...     

建立乳腺癌组织标本库标准化程序的初步研究

目的:探讨建立乳腺癌标本库的标准化程序,为建立系统的、规范的乳腺癌组织标本库及信息管理体系奠定基础.方法:收集乳腺癌患者术中的肿瘤组织、癌旁组织、远端正常组织标本,并收集部分患者血标本提取血清、血浆及淋巴细胞,所有组织标本均深低温保存.采用一套信息管理体系对于标本进行管理.结果:共收集22例乳腺癌患者新鲜组织及血样标本共520份,其中导管内癌2例,小叶原位癌3例,浸润性导管癌10例,浸润性小叶癌7例;全套标本者19例;标本留取时间平均为(19.38±10.16) min;随机抽取20份肿瘤组织提取DNA和RNA作质量监测,合格率达到90％.结论:建立系统的、规范的乳腺癌组织标本库及信息管理体系,将为乳腺癌的基础与临床研究提供物质与信息资源.     

强直性脊柱炎生物样本库的建立与质量控制

目的建立标准化及具有完整临床资料的强直性脊柱炎(AS)生物样本库,为AS的基础研究提供高质量的样本。方法自2009-06—2015-06共收集168例AS病例及93例对照病例(20例新鲜股骨颈骨折、42例股骨头坏死、20例髋臼发育不良及11例类风湿性关节炎),按照标准化的操作流程对入选患者术前血液、尿液和术中新鲜髋关节韧带组织标本进行收集、处理、分装、储存,对部分韧带组织行石蜡包埋保存。采用临床数据库软件及样本库管理软件建立临床资料完整的AS生物样本库,定期对样本进行质量监测。结果建立标准化的强直性脊柱炎及对照病例样本库,随机抽取AS病例样本及对照病例样本各3例送至第三方(上海其明生物公司)进行质量检测,各样本经Trizol试剂提取组织RNA,用分光光度仪测定A260/A280值,RNA样本经1%琼脂糖凝胶电泳,检测结果合格。结论建立的标准化AS生物样本库可为AS发病机制的研究提供高质量的样本。     

结直肠肿瘤生物样本库质量控制体系建立的经验与体会

目的通过创建结直肠肿瘤组织样本库质量控制体系,充分保护和合理利用结直肠肿瘤标本资源,为临床和科学研究者提供质量合格的结直肠肿瘤标本,促进结直肠肿瘤的研究。方法建立严格的结直肠肿瘤组织生物样本库质量控制体系,规范收集手术切除后的肿瘤组织,包括肿瘤、瘤旁和切缘远端正常组织以及术前、术后的外周血血清/血浆标本。建立一套完整的结直肠肿瘤组织提取、储存操作流程,制定质量控制方案,建立符合国际规范的结直肠肿瘤组织生物样本库及信息管理系统,提供临床样本资源。结果标本库现保存有完整临床资料及随访资料的结直肠肿瘤组织标本1600例。其中恶性肿瘤1225例,良性肿瘤275例。包括肿瘤组织、癌旁组织、远端正常组织标本以及术前、术后血液标本。完成了标本库信息数据管理软件的开发工作,对所收集标本的流行病学及临床资料信息录入计算机归档管理。结论初步建立了具有一定规模的肿瘤标本资源库,完善了结直肠肿瘤生物样本库质量控制体系,形成了一整套肿瘤组织标本库的建设及管理方法,为相关的结直肠肿瘤研究提供了服务平台。     

胆道系统肿瘤标本库的建立与分析

目的 建立胆道系统肿瘤血液及组织标本库,为进一步开展胆道肿瘤研究提供参考.方法 选取2010年3月至9月我院肝胆外科以胆道肿瘤为主要诊断而行手术切除的患者标本37例,留取血浆、血清及组织标本,制作石蜡包块并行常规病理组织学鉴定,并使用 Trizol 试剂提取总 RNA及基因组DNA.结果 37例标本中胆管癌18例,胆囊癌19例.两组间在性别、黄疸表现、是否合并胆道结石、能否根治性手术及近期生存时间存在差异.抽样检查显示77.8%的样本分子生物学指标稳定.结论 本实验建立的胆道肿瘤标本采集流程有效可行,临床资料收集完备,能够为今后胆道系统肿瘤的发生发展机制研究提供较好的标本资源.     

人脑胶质瘤组织库的建立及初步应用

我们通过收集脑胶质瘤组织建立肿瘤组织库,为胶质瘤的研究提供了资料来源. 一、资料与方法 1.胶质瘤组织库保存方法:新鲜肿瘤组织铝箔纸包裹液氮罐内长期保存.Trizol法进行总RNA提取后合成cDNA冻存.组织固定制成石蜡块常温保存.     

胆管癌新鲜组织标本库建立与鉴定

目的利用解放军总医院肝胆外科的医疗优势,摸索一套合理的操作方法,有计划地建立胆管癌新鲜组织标本库,为系统开展胆道肿瘤研究做好准备。方法选取2000年1月～2001年6月解放军总医院肝胆外科以胆道肿瘤为主要诊断而行手术切除的患者的手术标本21例,包括肿瘤组织、正常组织和(或)癌旁组织,将切取的组织块分割后装入编号的灭菌贮藏管,迅速放入液氮转移罐,深低温冰柜保存。全部组织标本均经常规病理组织学检查予以鉴定,并取其中部分组织使用Trizol试剂提取总RNA鉴定以判断组织成分的完整性。结果21例标本中肝内胆管癌4例,肝门胆管癌6例,肝外胆管癌7例,壶腹癌4例。由于肿瘤标本大小不同,所切取的组织块数目各异;因手术难度及人为因素的影响使得组织留取平均时间为(47.60±43.87)min。所取组织经Trizol试剂提取后,可见离心管底部沉淀的呈黄白薄片状的RNA,经紫外分光光度计分析得出其A260/A280=1.6～1.8,总RNA经1%琼脂糖凝胶电泳后可见明显的28s和18s条带,而且28s条带的亮度大约是18s的2倍;2例超过2h的标本的A260/A280<1.6,且电泳显示28s和18s条带不清、消失及拖尾。结论人类遗传资源库的建设是开展疾病相关基因研究的重要的基础性工作,本实验建立的胆管癌组织标本留取操作流程可保证标本库质量。标本的留取必须在知情同意和保证留有足够的用于病理诊断的组织前提下,由有经验的相对固定的医护人员完成;标本离体时间应掌握在1h之内,应常规将标本分割多管保存。     

无乙肝及丙肝的肝细胞癌病人肝组织中HBV DNA的重要性

无乙肝及丙肝的肝细胞癌其病原学和预后因素仍然不清楚。该研究通过原位杂交的方法，对46例无乙肝及丙肝的肝细胞癌病人的非肿瘤肝组织中HBV DNA和HCV RNA的水平进行检测。对HBV DNA高水平表达和HBV DNA低水平表达两组病人肿瘤再复发率进行比较，同时探索一些可能与预后有关的因素。结果显示，35例病人的非肿瘤肝标本中检测到HBV DNA，而所有标本中均没发现HCV RNA。[第一段]     

血液系统疾病生物样本库的建立

目的 探讨建立设施完备、信息互通的规范化血液系统疾病样本采集、存储、管理、服务体系,实现全方位血液系统活细胞及相关血液成分的规范化保藏,为临床研究及转化应用提供资源保障。方法 血液系统疾病生物样本库集收集处理、质量控制、基因分析、功能鉴定于一体,另设有独立的存储区,具有与临床诊疗数据、组学信息联通的一体化信息管理系统,以保藏血液系统疾病骨髓活细胞为特色,建立规范化样本接收、处理、存储及使用流程,定期进行质控检测,完善共享机制及运营模式。结果 血液样本资源库已保藏血液系统活细胞等各类样本共计超40万份,实现全部临床诊疗科室与疾病类型的样本全覆盖,并建立疾病来源i PSC七百余株。随机抽取新鲜和冻存样本6例,质量检测全部合格,为所院高质量科研产出提供样本支持。结论 通过全流程管控实现血液病理活细胞及相关组分的规范化收集、制备、存储和使用,建成以整合生物样本、临床诊疗数据、生物信息学等生物医学信息与资源为一体的“干湿”结合升级版生物样本库。     

肿瘤-睾丸抗原在胃癌组织中的表达

我们对MAGE 1、MAGE 3、BAGE、LAGE、NY ESO 1、SCP 1、SSX 2和SSX 4这 8种肿瘤 睾丸抗原 (CTA)在胃癌组织及其相应正常组织中的表达进行检测 ,旨在为特异性免疫治疗在胃癌中的应用提供依据。一、对象与方法1.组织标本 :42例胃癌组织标本取自上海瑞金医院普外科 2 0 0 3年 5月～2 0 0 3年 10月间手术治疗胃癌患者 ,同时取距离肿瘤边缘 5cm以上癌旁正常组织作为对照。标本经手术切除后立即放入液氮中 ,3 0min后保存于 -80℃直至RNA抽提。所有胃癌组织及相应正常组织均经术后病理证实。其中 ,男 2 8例 ,女 14例 ,低分化腺癌 2…     

Establishment of a biobank for human lung tissues of congenital diaphragmatic hernia and congenital pulmonary airway malformation

BackgroundHuman tissue samples are an invaluable and little available source of information for translational studies of congenital lung diseases such as Congenital Diaphragmatic Hernia (CDH) or Congenital Pulmonary Airway Malformation (CPAM).PurposeWe aimed to establish a human lung tissue biobank of CDH and CPAM patients together with age-matched controls, coupled with a clinical database.MethodsPathology records from autopsies or surgical specimens for CDH and CPAM cases between 1980 and 2017 were reviewed. For surviving individuals, clinical patient data was obtained from corresponding pediatric surgery reports. Formalin-fixed, paraffin-embedded tissues of patients and age-matched controls were systematically stored for further translational studies. RNA integrity was determined on selected CDH blocks.ResultsA total of 16 CDH and 18 CPAM and age-matched control lung tissue blocks were included in our biobank. Ages ranged from 22 to 41 weeks of gestation (GA) in CDH (33.9 ± 6.35 weeks) and 26 weeks (GA) and 12 years in CPAM (2.3 ± 3.7 y). RNA isolation from CDH and control blocks yielded good RNA quality (OD 260/280 ratio: 2.01–2.09, OD 260/230 ratio: 2.04–2.09).ConclusionWe established a unique human biobank for CDH and CPAM tissues. The combination with clinical patient data will allow us to design future translational studies to improve our understanding of the disease pathogenesis of these congenital malformations.     

胃癌根治术中组织印片细胞学检查的评价

目的 探讨印片细胞学检查在胃癌根治术中的快速诊断价值。方法 在52例胃癌根治术的新鲜标本上，分别于近端切缘、远端切缘、瘤组织、近瘤旁组织及部分淋巴结232处印片取材，凉干，行细胞学检查，观察有无瘤细胞。将其结果与常规病理检查结果相对照。结果 瘤组织处假阴性1例无假阳性；切缘处无假阴性，假阳性12例；瘤旁组织假阴性3例，假阳性10例；全组标本敏感性92．8％，特异性85．8％，准确性87．9％。结论 胃癌根治术中印片细胞学检查对判断切缘有无残留病灶有一定价值，尤其是对不具备快速病理检查的基层医院有帮助，但不能完全取代快速病理检查，二者结合更为有利。     

Use of bipolar energy for transurethral resection of bladder tumors: pathologic considerations

BACKGROUND AND PURPOSE: Bipolar electrocautery has recently been introduced as a modality for transurethral resection of bladder tumors (TURBT). The primary benefits of bipolar TURBT stem from the use of saline irrigant rather than glycine or water. TURBT should be conducted in a fashion such that the resected tissue can be used for proper grading and staging, so excessive cauterization of the tissue should be avoided. In this study, we compared the pathologic characteristics of bladder tumor specimens resected with bipolar versus standard monopolar energy to determine specimen quality. PATIENTS AND METHODS: Bipolar TURBT (Gyrus Medical Inc., Maple Grove, MN) was performed in 11 patients. Pathologic specimens were compared with the specimens from 11 patients who had previously undergone standard monopolar TURBT. Resected tissue was examined by a pathologist who recorded tumor size, grade, location, presence of muscularis propria, presence of muscle invasion, and final diagnosis. The pathologist also determined the degree of cautery artifact in each specimen. The pathologist was blinded to the form of electrocautery used and the clinical diagnosis. RESULTS: Transurethral resection with bipolar electrocautery was carried out without difficulty or complication in all cases. Similarly, there were no complications in resection by standard monopolar electrocautery. The bladder tumor chips obtained with bipolar TURBT were smaller because of the smaller size of the bipolar loop. However, this did not interfere with the pathologic assessment. There were no significant pathologic differences between specimens according to the type of cautery used. A large degree of cautery artifact was noted in the tissue of larger tumors resected using both monopolar and bipolar electrocautery. However, the incidence and degree of cautery artifact were similar in the two groups. No trends between tumor location and degree of cautery effect were noted. The pathologist had no difficulty reaching a full and proper diagnosis in all cases involving either form of electrocautery. CONCLUSIONS: Bipolar electrocautery is well suited for TURBT. Bladder tissue obtained from bipolar TURBT is of the same histologic quality as that obtained from standard monopolar TURBT and provides the urologist with a reliable and complete diagnosis.     

Biobanking of Fresh-frozen Human Colon Tissues: Impact of Tissue Ex-vivo Ischemia Times and Storage Periods on RNA Quality

Background

Biobanking plays an important role in translational cancer research. The impact of tissue ex-vivo ischemia time and storage period on RNA integrity is not well documented.

Methods

Fresh-frozen colon tissues were collected in Taizhou Hospital of Zhejiang Province in China since 2004. Fifty-one colon cancer tissues with tumor cell content higher than 70 % and matched normal tissues during four storage periods (less than 15 months, 16–20 months, 21–25 months, and 26–40 months) were chosen to detect RNA quality. Fresh colon cancer tissues from 5 patients were cut into pieces and kept at room temperature or on ice for 0.5, 1, 2, and 4 h before snap freezing. RNA integrity was determined by microcapillary electrophoresis by the RNA integrity number (RIN) algorithm.

Results

Sixty-seven percent of normal colon tissues and 94 % of colon cancer specimens yielded RNA with a RIN of ≥7. Matched colon cancer and normal tissues showed significant difference in RNA quality. RNA remained stable in colon cancer tissues kept at room temperature and on ice for up to 4 h, and long-term storage of banked colon specimens did not negatively influence RNA quality (RNA with RIN of ≥7 banked less than 15 months, 83 %; 16–20 months, 78 %; 21–25 months, 77 %; 26–40 months, 90 %).

Conclusions

Frozen colon tissues yield high-quality RNA in approximately 80 % of specimens. Ex-vivo ischemia times and storage periods did not adversely affect RNA quality. This study showed that standard operation protocols and the maintenance of high-quality tissue repositories were the keys to translational medicine research.     

DNA甲基转移酶在肝细胞癌中的表达及其临床意义

目的:探讨DNA甲基转移酶1(DNMT1)、DNA甲基转移酶3a(DNMT3a)和甲基转移酶3b(DNMT3b)在肝细胞癌(HCC)中的表达及其意义.方法:应用免疫组化检测47例HCC癌组织、42例HCC癌旁肝组织及12例正常肝组织中DNMT1、DNMT3a和DNMT3b的表达情况,分析三者与临床病理特征之间的关系.结果:DNMT1、DNMT3a和DNMT3b在HCC中阳性表达分别为80.9%、68.1%和78.7%,在癌旁组织内的表达分别为50.0%、52.4%和57.1%,均明显高于正常肝组织内的阳性表达(16.7%、16.7%和25.0%),且DNMT1、DNMT3a和DNMT3b与HCC的病理分化类型及乙肝表面抗原阳性显著相关(P<0.05).结论:DNMT1、DNMT3a和DNMT3b的异常表达与HCC的发生发展有紧密的关系.     

Polymerase chain reaction identification of human papillomavirus DNA in CO2 laser plume from recurrent respiratory papillomatosis.

Human papillomavirus (HPV) DNA was identified in the plume produced during CO2 laser vaporization of respiratory tract papillomata. The plume produced from CO2 vaporization was collected on Gelfoam pledgets that were affixed to suction tips evacuating the vapor plume from the operative field. The Gelfoam pledgets were snap frozen in liquid nitrogen, processed, and examined for HPV-6 and HPV-11 DNA by a polymerase chain reaction technique. Tissue and vapor-plume specimens were collected from 22 patients undergoing CO2 laser excision of laryngeal lesions. Seven patients had adult-onset recurrent respiratory laryngeal papillomatosis (RRP), 12 had juvenile-onset RRP, two had laryngeal carcinoma, and one had nonspecific laryngitis. HPV-6 or HPV-11 was identified in 17 of 27 vapor-plume specimens from RRP and in none of three from non-RRP lesions. All but one RRP tissue specimen contained HPV-DNA, and none of the non-RRP tissues contained HPV-DNA. When HPV was present in vapor, the same HPV type was found in the corresponding tissue specimen. Identification of HPV-DNA in the laser plume raises concern regarding potential risks from exposure to the plume--particularly to the endoscopic surgeon and the operating team. The practical concerns and effectiveness of the plume scavenging systems are discussed.     

Detection of telomerase activity in breast masses by fine-needle aspiration

Background: Telomerase is an RNA-dependent DNA polymerase that compensates for the telomere shortening that occurs in its absence. Reactivation of telomerase is thought to be an important step in cellular immortalization, and recent studies have indicated that telomerase activity is often detected in primary human malignancies. The clinical implications of telomerase activity in human tumors are currently under investigation. Methods: Eighty-nine samples (46 FNAs and 43 gross tissue biopsies) from 44 patients with breast masses were analyzed prospectively for the presence of telomerase activity by a modification of the telomere repeat amplification protocol (TRAP). All samples were obtained directly from the excised mass at the time of specimen removal in the operating room. Results: Telomerase activity was detected in 17 of 19 (90%) FNA samples and 15 of 18 (83%) invasive breast cancer tissue biopsies. Telomerase was also detected in 9 of 16 (56%) FNAs and 8 of 15 (53%) tissue biopsies from 16 fibroadenomas. Other benign proliferative lesions (n=5) did not have detectable telomerase activity in either FNA or tissue specimens. FNA-TRAP results correlated with the gross tissue specimen TRAP results in 95% of all cases. Conclusion: The FNA-TRAP assay for telomerase detection is a highly sensitive and accurate method for the detection of telomerase activity in breast masses. Future application of these techniques should facilitate evaluation of telomerase as a tumor marker in the clinical management of breast and other solid malignancies. These authors contributed equally to this work.