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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 *** are the largest available source of biological materials and are a potentially useful resource that can be used for retrospective epidemiological *** recovered from the slides can be used for several downstream molecular processes including polymerase chain reaction,single nucleotide polymorphism analysis,and whole genome *** DNA from these slides can be utilized to compare gene signatures of normal and diseased ***,extraction of high-quality DNA from archived stained hematoxylin and eosin(H&E)slides remains *** To standardize a new protocol for extracting DNA from archived H&E-stained tissue slides for further molecular *** A total of 100 archived H&E-stained cancer slides were subjected to a total of five methods of DNA *** were varied in the deparaffinization step,tissue rehydration,duration of lysis,and presence or absence of proteinase *** extracted DNA was quantified using a NanoDrop spectrophometer and the quality was analyzed by agarose gel *** 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 *** 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 *** yield was significantly higher when compared to other *** addition,90%of the extracted DNA showed amplifiable GAPDH *** 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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Standardized Assessment of Ki-67 in Breast Cancer Patients Using Virtual slides and an Automated Analyzer in Comparison to Central/Local Pathological Assessments

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Journal of Cancer Therapy 2014年第2期5卷 141-146页

作者： Yoshio Mizuno Hiromi Fuchikami Tsuneo Natori Naoko Takeda Yuko Inoue Junichi Yamada Hiroaki Abe Hiroshi Seto Kazuhiko Sato Clinical Testing Business Testing Division SRL Inc. Tokyo Japan Department of Breast Oncology Tokyo-West Tokushukai Hospital Tokyo Japan Department of Clinical Pathology Tokyo-West Tokushukai Hospital Tokyo Japan Inoue Ladies Clinic Tokyo Japan Seto Hospital Tokorozawa Japan

Purpose: To standardize the methods to measure Ki-67, there is an interest in automating the assessment of Ki-67. Therefore, we reviewed the possibility of introducing an automated analyzer to standardize the Ki-67 evaluation method. Methods: We retrospectively reviewed a clinical database of patients who underwent surgery for early breast cancer at Tokyo-West Tokushukai Hospital. Among them, those who underwent preoperative core needle biopsy (CNB) were enrolled. The concordance rates of estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER2), and Ki-67 by local pathologists were reviewed (valuations made by local pathologists), and nonmatching cases (from August 2008 to October 2011) were reassessed both by central review and using an automated analyzer with virtual slides. The results were compared with the evaluations made by local pathologists, and we reexamined the concordance rate by using central review and the automated analyzer. Results: The concordance rate of Ki-67 evaluations made by local pathologists in the preoperative CNB and surgical specimens was 78.7% in 287 cases pathologically assessed from October 2008 to March 2013. This rate was significantly lower (p < 0.01) than that of ER (95.6%), PgR (88.5%), and HER2 (91.6%). Reassessment of the 37 cases of nonmatching Ki-67 values from 2008 to October 2011 using central review and an automated analyzer resulted in clear improvement in matching of 22 (92.1%) and 24 (93.1%) of 37 cases, respectively. Conclusion: The concordance rate of Ki-67 in preoperative CNB and surgical specimens was lower than that of other biological markers;however, they were nearly equal by reassessment using central review and an automated analyzer.

关键词： Ki-67 Core Needle Biopsy Concordance Rate Virtual slides Automated Analyzer Standardized Assessment

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USING CYTOGENETIC ANALYSIS OF MOUSE EGGS FERTILIZED IN VITRO FOR MONITORING THE MUTAGENICITY OF CHEMICALS ACTING ON GERM CELLS——*** for preparation of slides of chromosome of mouse eggs at the first cleavage metaphase and the preliminary use

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癌变．畸变．突变 1991年第S1期3卷 116-116页

In this experiment,the sperms were obtained from(C57×CBA) F1 male mice and the eggs from NIHstrain female *** the fertilization techni-que in Vitro,the slides of chromosome of mouse eggsat the first cleavage metaphas... 详细信息

关键词： chromosome slides preliminary cleavage fertilization Chromosome sperm incubation mitomycin separate

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Integration of InSAR and GPS for hydraulic engineering

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Science China(Technological Sciences) 2007年第Z1期50卷 111-124页

作者： HE XiuFeng LUO HaiBin HUANG QiHuan HE Min Institute of Satellite Navigation & Spatial Information System Hohai UniversityNanjing 210098China Institute of Satellite Navigation & Spatial Information SystemHohai UniversityNanjing 210098China Institute of Satellite Navigation & Spatial Information SystemHohai UniversityNanjing 210098China Institute of Satellite Navigation & Spatial Information SystemHohai UniversityNanjing 210098China

Interferometric synthetic aperture radar (InSAR) is a potential earth observation approach, and it has been demonstrated to have a variety of applications in measuring ground movement, urban subsidence and landslides. Currently InSAR provides the ability to map accurate DEM and measure ground deformation to sub-centimeter accuracy. However, many factors affect InSAR to measure ground movement since dam constructions are built in a large scale area with a complicated climate and unstable geology. This paper discusses potential applications of integrated InSAR and GPS to monitor a large-scale ground movement due to hydropower developments. The integration of InSAR and GPS can provide a cost-effective means for monitoring deformation of hydropower developments. Moreover, two novel methods, both the improved spatial interpolating method and estimation of 3D surface motion velocities method, are proposed and the experimental results and analysis are given in this paper.

关键词： InSAR GPS hydraulic and hydropower engineering slides monitoring

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The landslide problem

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Journal of Palaeogeography 2015年第2期4卷 109-166页

作者： G.Shanmugam Department of Earth and Environmental Sciences The University of Texas at Arlington

The synonymous use of the general term ＂landslide＂, with a built-in reference to a sliding motion, for all varieties of mass-transport deposits （MTD）, which include slides, slumps, debrites, topples, creeps, debris avalanches etc. in subaerial, sublacustrine, sub- marine, and extraterrestrial environments has created a multitude of conceptual and nomen- clatural problems. In addition, concepts of triggers and Iong-runout mechanisms of mass movements are loosely applied without rigor. These problems have enormous implications for studies in process sedimentology, sequence stratigraphy, palaeogeography, petroleum geol- ogy, and engineering geology. Therefore, the objective of this critical review is to identify key problems and to provide conceptual clarity and possible solutions. Specific issues are the fol- lowing：（1） According to ＂limit equilibrium analyses＂ in soil mechanics, sediment failure with a sliding motion is initiated over a shear surface when the factor of safety for slope stability （F） is less than 1. However, the term landslide is not meaningful for debris flows with a flowing mo- tion. （2） Sliding motion can be measured in oriented core and outcrop, but such measurement is not practical on seismic profiles or radar images. （3） Although 79 MTD types exist in the geological and engineering literature, only slides, slumps, and debrites are viable depositional facies for interpreting ancient stratigraphic records. （4） The use of the term landslide for high- velocity debris avalanches is inappropriate because velocities of mass-transport processes cannot be determined in the rock record. （5） Of the 21 potential triggering mechanisms of sediment failures, frequent short-term events that last for only a few minutes to several hours or days （e.g., earthquakes, meteorite impacts, tsunamis, tropical cyclones, etc.） are more relevant in controlling deposition of deep-water sands than sporadic long-term events that last for thousands to millions of years （e.g.,

关键词： debris flows landslides mass-transport deposits (MTD) slides slumps soilstrength triggering mechanisms reservoir characterization

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LIGHT DEMANDS OF JUVENILE LAMINARIA JAPONICA

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Chinese Journal of Oceanology and Limnology 1989年第1期7卷 1-9,97页

作者：费修绠蒋本禹丁美丽吴雅景黄如意李汉昌 Sansha Lamnaria Sporeling Cultivation Station Fujian

The light demands of seaweeds is an interesting and rather complex phenomenon because they depend not only on the species but also on their different development stages. Even different parts of the same plant sometimes have different light demands. Light control is an important procedure at large scale Lamnaria nursery stations in China. Technicians and scientists have different viewpoints on the best method to regulate light. A culture study on Lamnaria japonica starting from zoospores to several centimeter sporophytes to find the optimal and critical irradiance ranges for juvenile Laminaria at different development stages added more knowledge on this aspect. Experiment results show gametophytes can not tolerate irradiance of more than 150uEm-2s-1 while sporophytes can tolerate more than 519 uEm-2s-1. This big difference starts from the very early stage of 1-to 2-celled sporophytes. The biological basis and mechanism of this phenomenon need further research.

关键词： Laminaria juvenile strings sometimes starts cultivation slides young regulate naked

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Generation and Propagation of Tsunami by a Moving Realistic Curvilinear Slide Shape with Variable Velocities in Linearized Shallow-Water Wave Theory

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Engineering（科研） 2010年第7期2卷 529-549页

作者： Hossam Shawky Hassan Khaled Tawfik Ramadan Sarwat Nageeb Hanna 不详

The process of tsunami evolution during its generation under the effect of the variable velocities of realistic submarine landslides based on a two-dimensional curvilinear slide model is investigated. Tsunami generation from submarine gravity mass flows is described in three stages. The first stage represented by a rapid curvilinear down and uplift faulting with rise time. The second stage represented by a unilaterally propagation in the positive x direction to a significant length to produce curvilinear two-dimensional models represented by a depression slump, and a displaced accumulation slide model. The last stage represented by the time variation in the velocity of the accumulation slide (block slide). By using transforms method, Laplace in time and Fourier in space, tsunami waveforms within the frame of the linearized shallow water theory for constant water depth are analyzed analytically for the movable source model. Effect of the water depths on the amplification factor of the tsunami generation by the submarine slump and slide for different propagation lengths and widths has been studied and the results are plotted. Comparison of tsunami peak amplitudes is discussed for different propagation lengths, widths and water depths. In addition, we demonstrated the tsunami propagation waveforms after the slide stops moving at different propagation times.

关键词： Near-Far Field Tsunami Amplitudes Shallow Water Theory Water Wave Submarine Slumps and slides Laplace and Fourier Transforms

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EFFECT OF BIMOLANE (AT-1727) ON THE CELL CYCLE OF L1210 LEUKEMIA CELLS IN MICE

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Chinese Science Bulletin 1986年第18期31卷 1285-1290页

作者：张覃沐林晨王绵英 Department of Pharmacology Henan Medical Institute Zhengzhou Department of Pharmacology Henan Medical Institute Zhengzhouimolane (AT-1727) a new antitumor drug was synthesized by Prof. Ren Yun-feng of the Shanghai Institute of Materia Medica Academia Sinica. It is a derivative of diketopiperazine with two symmetrical morpholinmethyl groups attached to ICRF-

Bimolane (AT-1727), a new antitumor drug, was synthesized by Prof. Ren Yun-feng of the Shanghai Institute of Materia Medica, Academia Sinica. It is a derivative of diketopiperazine with two symmetrical morpholinmethyl... 详细信息

关键词： symmetrical derivative attached CELL antitumor slides noted eared totally plateau

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FAULT ROCKS AND SOUTHWARD THRUSTING OF DABIE COMPLEX IN EAST DABIE MOUNTAINS, ANHUI, CHINA

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Chinese Science Bulletin 1984年第11期29卷 1527-1530页

作者：徐树桐董树文周海渊陈冠宝 Geological Institute of Anhui Province Hefei

The essential part of Dabie Complex in the area discussed is composed of different kinds of fault rocks produced by two syntectonic slides during southward movements of Dabie Massif; some post tectonic ones, namely, t... 详细信息

关键词： tectonic slides EAST strata thrust brittle migrate mantle subdivided marginal

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Infectious heart disease

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China Medical Abstracts(Internal Medicine) 2007年第4期24卷 203-204页

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