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DNA extraction from archived hematoxylin and eosin-stained tissue slides for downstream molecular analysis

World Journal of Methodology 2019年第3期9卷 32-43页

作者： Pushkal Sinduvadi Ramesh Venkatesh Madegowda Suprith Kumar Shailashree Narasimha Parichay S R Nandini Nandish Manoli Devananda Devegowda Center of Excellence in Molecular Biology and Regenerative Medicine Department of BiochemistryJSS Medical CollegeJSS Academy of Higher Education and ResearchMysuru 570015India CIPHER Healthcare Pvt Ltd. Hyderabad 500034India Department of Pathology JSS Medical CollegeJSS Academy of Higher Education and ResearchMysuru 570015India

BACKGROUND Histopathologically stained archived tissue slides are stored in hospital archives for years to decades.They are the largest available source of biological materials and are a potentially useful resource that can be used for retrospective epidemiological studies.DNA recovered from the slides can be used for several downstream molecular processes including polymerase chain reaction,single nucleotide polymorphism analysis,and whole genome sequencing.The DNA from these slides can be utilized to compare gene signatures of normal and diseased tissues.However,extraction of high-quality DNA from archived stained hematoxylin and eosin(H&E)slides remains challenging.AIM To standardize a new protocol for extracting DNA from archived H&E-stained tissue slides for further molecular assays.METHODS A total of 100 archived H&E-stained cancer slides were subjected to a total of five methods of DNA extraction.Methods were varied in the deparaffinization step,tissue rehydration,duration of lysis,and presence or absence of proteinase K.The extracted DNA was quantified using a NanoDrop spectrophometer and the quality was analyzed by agarose gel electrophoresis.Then each sample was subjected to polymerase chain reaction(PCR)to amplify the internal control gene GAPDH,thereby confirming the DNA intactness,which could be further utilized for other downstream applications.RESULTS Of the five different methods tested,the third method wherein xylene was used for tissue deparaffinization followed by 72 h of digestion and without proteinase K inactivation yielded the highest amount of DNA with good purity.The yield was significantly higher when compared to other methods.In addition,90%of the extracted DNA showed amplifiable GAPDH gene.CONCLUSION Here we present a step-by-step,cost-effective,and reproducible protocol for the extraction of PCR-friendly DNA from archived H&E-stained cancer tissue slides that can be used for further downstream molecular applications.

关键词： DNA extraction hematoxylin and eosin tissue slides Molecular analysis Polymerase chain reaction Deparaffinization

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Feasibility of terahertz imaging for discrimination of human hepatocellular carcinoma

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World Journal of Gastrointestinal Oncology 2019年第2期11卷 153-160页

作者： Feng Duan Yu-Ye Wang De-Gang Xu Jia Shi Lin-Yu Chen Li Cui Yan-Hua Bai Yong Xu Jing Yuan Chao Chang Department of Interventional Radiology the General Hospital of Chinese People’s Liberation Army Institute of Laser and Optoelectronics School of Precision Instruments and Optoelectronic Engineering Key Laboratory of Optoelectronic Information Science and Technology (Ministry of Education) Tianjin University Department of Hepatobiliary and Pancreatic Surgical Oncology the General Hospital of Chinese People’s Liberation Army Department of Pathology the General Hospital of Chinese People’s Liberation Army Advanced Interdisciplinary Technology Research Center National Innovation Institute of Defense Technology

BACKGROUND Hepatocellular carcinoma(HCC) is one of the most common malignant tumors worldwide, and novel methods for early/rapid diagnosis of HCC are needed.Terahertz(THz) spectroscopy is considered to have the potential to distinguish between normal liver tissue and HCC tissue; however, there are few reports on it.We conduct this observational study to explore the feasibility of THz imaging for the diagnosis of HCC.AIM To evaluate the feasibility of THz for discriminating between HCC and normal liver tissues using fresh tissue specimens obtained from HCC patients who had undergone surgery.METHODS Normal liver tissue and HCC tissue were cryosectioned into 50 μm-thick slicesand placed on cover glass. Two adjacent tissue sections were separated subjected to histopathological examination by hematoxylin and eosin staining or THz transmission examination, and THz images were compared with pathologically mapped images. We determined the typical tumor and normal liver tissue regions by pathological examination; the corresponding areas of adjacent sections were examined by THz transmission.RESULTS The transmission rate of HCC tissue was 0.15-0.25, and the transmission rate of typical HCC tissue was about 0.2. THz transmittance in normal liver tissue is slightly higher than 0.4, but there were many influencing factors, including the degree of liver cirrhosis, fat components, ice crystals in frozen sections, and apoptosis.CONCLUSION In conclusion, this study shows that THz imaging can detect HCC tissue. Further research will yield more detailed data of the THz transmission rates of HCC tissue with different degrees of differentiation.

关键词： Terahertz imaging Hepatocellular carcinoma hematoxylin and eosin staining Terahertz transmittance Liver pathology

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Clinical significance of cytomegalovirus infection in patients with inflammatory bowel disease

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World Journal of Gastroenterology 2013年第1期19卷 17-25页

作者： Elena Garrido Elisa Carrera Rebeca Manzano Antonio Lopez-Sanroman Inflammatory Bowel Disease Clinic Department of GastroenterologyUniversity Hospital Ramón y Cajal28034 MadridSpain Department of Gastroenterology University Hospital of Guadalajara19002 GuadalajaraSpain Department of Gastroenterology Hospital Sureste28500 MadridSpain

Cytomegalovirus(CMV) infection is common in humans.The virus then enters a "latency phase" and can reactivate to different stimuli such as immunosuppression.The clinical significance of CMV infection in inflammatory bowel disease is different in Crohn's disease(CD) and ulcerative colitis(UC).CMV does not interfere in the clinical course of CD.However,CMV reactivation is frequent in severe or steroid-resistant UC.It is not known whether the virus exacerbates the disease or simply appears as a bystander of a severe disease.Different methods are used to diagnose CMV colitis.Diagnosis is classically based on histopathological identification of viral-infected cells or CMV antigens in biopsied tissues using haematoxylin-eosin or immunohistochemistry,other tests on blood or tissue samples are currently being investigated.Polymerase chain reaction performed in colonic mucosa has a high sensitivity and a positive result could be associated with a worse prognosis disease;further studies are needed to determine the most appropriate strategy with positive CMV-DNA in colonic mucosa.Specific endoscopic features have not been described in active UC and CMV infection.CMV colitis is usually treated with ganciclovir for several weeks,there are different opinions about whether or not to stop immunosuppressive therapy.Other antiviral drugs may be used.Multicenter controlled studies would needed to determine which subgroup of UC patients would benefit from early antiviral treatment.

关键词： Cytomegalovirus Inflammatory bowel disease Crohn’s disease Ulcerative colitis hematoxylin and eosin Immunohistochemical Polymerase chain reaction Ganciclovir Infectious colitis

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RELATION BETWEEN DISTRIBUTION OF ATRIOPEPTIN IMMUNOREACTIVE MYOCYTES AND STRUCTURAL CHANGES OF LEFT VENTRICULAR WALL IN PRENATAL AND POSTNATAL RATS

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Chinese Science Bulletin 1989年第7期34卷 615-616页

作者：纪中生李肇特 Department of Histology and Embryology Beijing Medical University Beijing

We have reported that parts of myocytes in ventricle of the rat embryos show atriopeptin immunoreactivity. In this letter the relationship between the structural evolution of left ventricular wall and the distribution... 详细信息

关键词： letter RELATION ventricle hematoxylin neonatal histology luminal remaining transformed sections

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Diagnostic value of endothelial markers and HHV-8 staining in gastrointestinal Kaposi sarcoma and its difference in endoscopic tumor staging

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World Journal of Gastroenterology 2013年第23期19卷 3608-3614页

作者： Naoyoshi Nagata Toru Igari Takuro Shimbo Katsunori Sekine Junichi Akiyama Yohei Hamada Hirohisa Yazaki Norio Ohmagari Katsuji Teruya Shinichi Oka Naomi Uemura Department of Gastroenterology and Hepatology National Center for Global Health and MedicineTokyo 162-8655 Japan Department of Clinical pathology National Center for Global Health and MedicineTokyo 162-8655 Japan Department of Clinical Research and Informatics National Center for Global Health and MedicineTokyo 162-8655 Japan Division of AIDS Clinical Center (ACC) National Center for Global Health and MedicineTokyo 162-8655 Japan Department of Infectious disease National Center for Global Health and MedicineTokyo 162-8655 Japan Department of Gastroenterology and Hepatology National Center for Global Health and Medicine Kohnodai HospitalChiba 272-8516 Japan

AIM: To clarify the diagnostic values of hematoxylin and eosin (HE), D2-40, CD31, CD34, and HHV-8 immunohistochemical (IHC) staining in gastrointestinal Kaposi's sarcoma (GI-KS) in relation to endoscopic tumor staging. METHODS: Biopsy samples (n = 133) from 41 human immunodeficiency virus-infected patients were reviewed. GI-KS was defined as histologically negative for other GI diseases and as a positive clinical response to KS therapy. The receiver operating characteristic area under the curve (ROC-AUC) was compared in relation to lesion size, GI location, and macroscopic appearances on endoscopy. RESULTS: GI-KS was confirmed in 84 lesions (81.6%). Other endoscopic findings were polyps (n = 9), inflammation (n = 4), malignant lymphoma (n = 4), and condyloma (n = 2), which mimicked GI-KS on endoscopy. ROC-AUC of HE, D2-40, blood vessel markers, and HHV-8 showed results of 0.83, 0.89, 0.80, and 0.82, respectively. For IHC staining, the ROC-AUC of D2-40 was significantly higher (P < 0.05) than that of HE staining only. In the analysis of endoscopic appearance, the ROC-AUC of HE and IHC showed a tendency toward an increase in tumor staging (e.g. , small to large, patches, and polypoid to SMT appearance). D2-40 was significantly (P < 0.05) advantageous in the upper GI tract and for polypoid appearance compared with HE staining. CONCLUSION: The diagnostic value of endothelial markers and HHV-8 staining was found to be high, and its accuracy tended to increase with endoscopic tumor staging. D2-40 will be useful for complementing HE staining in the diagnosis of GI-KS, especially in the upper GI tract and for polypoid appearance.

关键词： Gastrointestinal Kaposi’s sarcoma hematoxylin and eosin CD31 CD34 D2-40 Human herpesvirus-8

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甜菜(Beta Vulgaris)染色体苏木精(hematoxylin)分化染色简报

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甜菜糖业 1982年第1期 10-10页

作者：郭德林李山源白庆武贾树彪金炳浩张启荣温祥廉永云齐齐哈尔师范学院生物系轻工业部甜菜糖业研究所

研究甜菜核型,从形态特征上鉴别每对染色体,是甜菜遗传学和育种实践的重大课题。但甜菜与大多数重要的经济作物不同,染色体在形状、大小、着丝点指数等指标上,极为相近,六十年的研究成果,仅可有效地鉴别第1、第2和第9染色体。近年,一些... 详细信息

研究甜菜核型,从形态特征上鉴别每对染色体,是甜菜遗传学和育种实践的重大课题。但甜菜与大多数重要的经济作物不同,染色体在形状、大小、着丝点指数等指标上,极为相近,六十年的研究成果,仅可有效地鉴别第1、第2和第9染色体。近年,一些学者开展甜菜中期染色体Giemsa分带,目的在于揭示并定位染色体的结构差异。

关键词：染色体长臂遗传学 Beta Vulgaris hematoxylin 苏木精甜菜糖料作物

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Primary Cutaneous Gamma Delta T Cell Lymphoma: A Clinicopathological Analysis

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Journal of Biosciences and Medicines 2019年第5期7卷 22-26页

作者： Li Tang Yu Li Department of Pathophysiology Chongqing Medical University Chongqing China Department of Pathology Institute of Neuroscience Chongqing Medical University Chongqing China

Primary cutaneous gamma-delta T-cell lymphoma (PCGD-TCL) is a very uncommon and extremely aggressive tumor, and is described in the newly re-vised World health organization for research and treatment of cancer classification of cutaneous lymphomas. A 43-year-old male patient presented with a 4 months history of cutaneous lesion over upper lip, without plaque and any constitutional symptom. Histopathological examination of skin biopsy revealed infiltration of atypical lymphocytes with hyperchromatic irregular nuclei. Immunophenotyping pattern of skin biopsy was compatible with PCGD-TLC. It is a highly aggressive tumor resistant to chemotherapy, immunotherapy, and radiation therapy. The GDTCL is characterized by a worse prognosis with a median survival of 15 months. Early diagnosis is essential and aggressive therapy is necessary.

关键词： Cutaneous Gamma-Delta T-Cell Lymphoma hematoxylin and Eosin Immunohistochemistry

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Role of Immunostaining in Detecting Extra-Pattern and Subtle Lymphomatous Infiltration in Bone Marrow Biopsies of NHL Patients

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Open Journal of Blood Diseases 2018年第2期8卷 27-36页

作者： Noha Bassiouny Hassan Yasmin N. El Sakhawy Gehan M. Hamed Department of clinical Pathology Faculty of Medicine Ain Shams University Hospital Cairo Egypt

Introduction: Immunohistochemistry (IHC) enables the examination of a greater number of trephine biopsy levels and is helpful in determining additional scattered malignant cells. The aim of this study is to detect extra-pattern and subtle lymphomatous infiltration in bone marrow biopsies using CD20 and CD3 immunostaining. Patients and Methods: This study was conducted on 100 newly diagnosed Non Hodgkin Lymphoma (NHL) patients. Their bone marrow trephine biopsies were assessed on routine histology [hematoxylin and Eosin (H & E)], and were further subjected to IHC using CD20 and CD3. Results: Pattern of involvement by H & E was highlighted by IHC. It showed additional interstitial pattern in 9 cases, parasinusoidal streaks in one case and highlighted a patchy pattern in another case with interstitial involvement on H & E. IHC also detected subtle infiltrations on additional 5.5% cases compared with histology alone. It helped in differentiating reactive (12 cases) and malignant lymphoid infiltration (33 cases). Conclusion: CD20 and CD3 immunostaining performed routinely on bone marrow trephine biopsies has the ability to reveal extra-pattern of infiltration and improve detection of subtle lymphoid involvement. A combined procedure identifying several distinctive features, in particular histotopography and IHC, provides a promising way of discriminating reactive from neoplastic lymphoid infiltrates in bone marrow trephine biopsies.

关键词： Immunohistochemistry (IHC) hematoxylin and Eosin (H & E) Non-Hodgkin Lymphoma (NHL)

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Effects of 1,25( OH)_2D_3 on bleomycin-induced pulmonary fibrosis in mice

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China Medical Abstracts(Internal Medicine) 2014年第1期31卷 32-33页

作者：张宗梅 Dept Pathol Shanghai Tongji HospTongji Univ Med Sch

Objective To observe the effects of 1,25(OH)2D3on bleomycin-induced pulmonary fibrosis in mice and to explore its mechanisms.Methods Ninety male C57BL/6mice,6 to 8 weeks old,were randomly divided into 3groups according to the table of random numbers:a control group,a model group and a treatment group(n=30each).Bleomycin was injected to the mice in the latter 2groups by single intratracheal injection to duplicate the pulmonary fibrosis model,while the control group was injected

关键词： injected duplicate latter hydroxyproline hematoxylin killed collagen saline olive catenin

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Pathological changes of the livers from 39 patients with hepatic failure

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China Medical Abstracts(Internal Medicine) 2006年第4期23卷 219-220页

作者：崇雨田 Dept Infect 3rd Affili Hosp SUN Yat-sen Univ Guangzhou 510630

Objective To explore the pathological changes of the livers from hepatic failure (HF) patients and its association with clinical disease stages. Methods Thirtynine patients with liver failure caused by HBV infections were investigated, and none accompanied with hepatocellular carcinoma. The sections of tissue were taken from the liver after liver transplantation and stained with hematoxylin -eosin (H&E) or RT (reticular fiber) staining. The pathological features were analyzed and compared between the clinical and pathological diagnosis.

关键词： hematoxylin stained sections eosin accompanied staining sociation

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