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Yuexing Gu,1,2,* Yan Li,3,* Shengjie Ge,1,2 Wenbo Lu,4 Yu Mao,1,2 Miao Chen,1,2 Yayun Qian1,2 1Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, People’s Republic of China; 2Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University, Yangzhou, People’s Republic of China; 3Department of Obstetrics and Gynecology, The Second People’s Hospital of Taizhou City, Taizhou, People’s Republic of China; 4Shanxi Normal University, College of Chemistry and Material Science, Linfen, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yayun Qian, Email yyqian@yzu.edu.cnPurpose: This paper aims to construct a surface-enhanced Raman spectroscopy (SERS) biosensor based on functionalized Au-Si nanocone arrays (Au-SiNCA) using a dual signal amplification strategy (SDA-CHA) to evaluate telomerase activity during epithelial-mesenchymal transition (EMT) in laryngeal carcinoma (LC).Methods: A SERS biosensor based on functionalized Au-SiNCA was designed with an integrated dual-signal amplification strategy to achieve ultrasensitive detection of telomerase activity during EMT in LC patients.Results: Labeled probes (Au-AgNRs@4-MBA@H1) and capture substrates (Au-SiNCA@H2) were prepared by modifying hairpin DNA and Raman signal molecules. Using this scheme, telomerase activity in peripheral mononuclear cells (PMNC) could be successfully detected with a limit of detection (LOD) as low as 10− 6 IU/mL. In addition, biological experiments using BLM treatment of TU686 effectively mimicked the EMT process. The results of this scheme were highly consistent with the ELISA scheme, confirming its accuracy.Conclusion: This scheme provides a reproducible, selective, and ultrasensitive assay for telomerase activity, which is expected to be a potential tool for the early screening of LC in future clinical applicati

Yayun Qian,1 Yuexing Gu,2 Jialin Deng,3 Zhaoying Cai,3 Yang Wang,3 Ruoyu Zhou,3 Dongxu Zhu,3 Hongmei Lu,4 Zheng Wang1 1Department of Pathology, The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou, People’s Republic of China; 2Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, People’s Republic of China; 3Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University, Yangzhou, People’s Republic of China; 4Department of Pathology, Yangzhou Maternal and Child Health Hospital, Yangzhou, People’s Republic of ChinaCorrespondence: Zheng Wang; Yayun Qian, Email yzdxfsyywz@163.com; yyqian@yzu.edu.cnIntroduction: As the most common malignant tumor in the world, the prognosis of patients with advanced lung cancer remains poor even after treatment. There are many prognostic marker assays available, but there is still more room for the development of high-throughput and sensitive detection of circulating tumor DNA (ctDNA). Surface-enhanced Raman spectroscopy (SERS), a spectroscopic detection method that has received wide attention in recent years, can achieve exponential amplification of Raman signals by using different metallic nanomaterials. Integrating SERS with signal amplification strategy into the microfluidic chip and applying it to ctDNA detection is expected to be an effective tool for the prognosis of lung cancer treatment effect in the future.Methods: To construct a high-throughput SERS microfluidic chip integrated with enzyme-assisted signal amplification (EASA) and catalytic hairpin self-assembly (CHA) signal amplification strategies, using hpDNA-functionalized Au nanocone arrays (AuNCAs) as capture substrates and cisplatin-treated lung cancer mice to simulate the detection environment for sensitive detection of ctDNA in serum of lung cancer patients after treatment.Results: The SERS microfluidic chip constructed by