Methods in molecular biology
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The simple applicability and facile target programming of the CRISPR/Cas9-system abolish the major boundaries of previous genome editing tools, making it the tool of choice for generating site-specific genome alterations. Its versatility and efficacy have been demonstrated in various organisms; however, accurately predicting guide RNA efficiencies remains an organism-independent challenge. Thus, designing optimal guide RNAs is essential to maximize the experimental outcome. Here, we summarize the current knowledge for guide RNA design and highlight discrepancies between different experimental systems.
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Surgery is the mainstay treatment for esophageal squamous cell carcinoma (ESCC). A major development in surgical therapy for esophageal cancer in the past two or three decades has been the marked reduction in surgical mortality rates as a result of improvement in staging technologies, patient selection, perioperative support systems, and surgical experience. Minimally invasive techniques are also becoming widely adopted. Protocol for surgical management of squamous cell cancer of the esophagus is described herein.
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MicroRNAs (miRNAs) are 20-22 nucleotides long single-stranded noncoding RNAs. They regulate gene expression posttranscriptionally by base pairing with the complementary sequences in the 3'-untranslated region of their targeted mRNA. Aberrant expression of miRNAs leads to alterations in the expression of oncogenes and tumor suppressors, thereby affecting cellular growth, proliferation, apoptosis, motility, and invasion capacity of gastrointestinal cells, including cells of esophageal squamous cell carcinoma (ESCC). ⋯ Consequently, expression profiles of miRNAs could be useful as diagnostic, prognostic, and prediction biomarkers in ESCC. Herein, we describe the quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and microarray methods for detection and quantitate miRNAs in ESCC. In addition, we summarize the roles of miRNAs in ESCC pathogenesis, progression, and prognosis.
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Macroscopic examination of the surgical specimen of esophageal squamous cell carcinoma by pathologist is important for quality clinical management, research, as well as education purposes. The process includes dissection of the specimen, identification of the lesion, measurements, and taking appropriate samples for histopathological examination. ⋯ In addition, examination of proximal resection margin and circumferential resection margin are important in the cancer. A standardized approach for macroscopic examination by professionals is needed for accurate diagnosis and to optimize the use of the surgical specimen with esophageal squamous cell carcinoma.
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Whole-slide imaging (WSI) contributes to medical education, collaboration, quality assurance, examination, and consultation in pathology. The images obtained from WSI are of high quality and could be stored indefinitely. ⋯ The operation and basic principles of whole-slide imaging of esophageal squamous cell carcinoma are also presented. Common use of WSI will occur with modifications of the whole-slide imaging scanners to adapt to the workflows in diagnostic and research laboratories.