Methods in molecular biology
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While surgery plays a major role in the treatment and potential cure of esophageal cancers, esophagectomy remains a high-risk operation with significant perioperative morbidity and mortality compared to other oncosurgical procedures. Perioperative management for esophagectomy is complex, and close attention to detail in various areas of anesthetic and perioperative management is crucial to improve postoperative outcomes. Patients undergoing esophagectomy should be offered an evidence-based risk assessment for their postoperative outcomes to allow active participation and informed, shared-decision making. ⋯ Furthermore, anesthetic practice and perioperative anesthetic drug usage can potentially affect cancer progression and recurrence. This chapter reviews current evidence for various factors that contribute to the improvement of perioperative outcomes, including prehabilitation, preoperative optimization of anemia, thoracic epidural analgesia, intraoperative protective ventilatory strategies, goal-directed fluid therapy, as well as special attention to other perioperative issues that potentially reduce anastomotic and cardiopulmonary complications. In summary, it is difficult to show a measurable benefit from any one single intervention, and a multidisciplinary approach that encompasses multiple aspects of perioperative care is necessary to improve outcomes after esophagectomy.
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Tissue microarray (TMA) is widely used for identifying the expression of markers in many tissues from patients with esophageal squamous cell carcinoma. The technology is mostly used in immunohistochemical studies to test the expression of markers and oncoproteins in signalling pathway as well as targeting proteins involved in therapies for esophageal squamous cell carcinoma. Appropriate use of TMA sections needs consideration of labor, planning, and expertise involved. For the best performance, it is important to design the layout of the TMA as well as use whole-slide scanning for interpretation of the TMA sections.
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Cervical esophageal carcinoma (CEC) is rare, accounting for 2-10% of esophageal cancers and is mostly squamous cell carcinoma. Because of the anatomical proximity of CEC to larynx, surgical treatment would involve pharyngo-laryngo-esophagectomy (PLE) with inherent high mortality and morbidity. Laryngeal preservation is an important consideration, and definitive chemoradiotherapy is the recommended treatment. ⋯ Since the exact location, extent of primary and nodal metastasis varies between patients, radiotherapy treatment needs to be individualized. The optimal radiation dose for CEC is uncertain, but retrospective data suggests that higher radiation dose of at least 60 Gy is associated with better local control and survival. Advanced radiotherapy technique, like intensity modulated radiotherapy, is usually required to achieve high dose to tumor while protecting normal tissues from excessive radiation.
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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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Technological advances in the form of next-generation sequencing allow sequencing of large numbers of different DNA sequences in a single/parallel reaction compared to conventional sequencing. It is a powerful tool which has enabled comprehensive characterization of esophageal squamous cell carcinoma. Whole-genome sequencing is the most comprehensive but expensive, whereas whole-exome sequencing is cost-effective, but it only works for the known genes. Thus, second-generation sequencing methods can provide a complete picture of the esophageal squamous cell carcinoma genome by detecting and discovering different type of alterations in the cancer which may lead to the development of effective diagnostic and therapeutic approaches for esophageal squamous cell carcinoma.