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However, current clinical practice lacks a reliable method for real-time AzoR monitoring to effectively guide personalized therapies and assess treatment responses. We address this challenge by developing an AzoR-activatable near-infrared (NIR) fluorescent self-assembled probe, NFK-(Azo)-DCM, which combines self-assembly properties with hypoxia-specific activation through a rationally designed azobenzene unit. This molecular architecture enables a selective chemical transformation\u2500the conversion of azobenzene to an electron-donating amino group under AzoR overexpression in hypoxic tissues\u2500which initiates a cascade of NIR fluorescence signal amplification. Confocal imaging revealed that NFK-(Azo)-DCM exhibited statistically significant (P &amp;lt; 0.01) fluorescence enhancement in LLC, 4T1, and HEK293 cells under hypoxic conditions. In LLC tumor-bearing mice, the probe produced a 3.5-fold increase in the NIR fluorescence signal specifically in hypoxic tumor regions compared to the negative controls, establishing its utility for tumor bioimaging. Most notably, in a murine hindlimb ischemia model, the probe achieved rapid and accurate detection of AzoR activity through its NIR emission, with a strong linear correlation between the signal intensity and ischemic severity (R2 &amp;gt; 0.9). The diagnostic versatility of NFK-(Azo)-DCM was further validated in both the acute and chronic UC models. These findings establish NFK-(Azo)-DCM as a dual-functional platform capable of noninvasive, real-time monitoring of hypoxia-related diseases, providing a promising method for advancing research in AzoR-associated pathologies.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c02438","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T13:03:25Z","timestamp":1786539805000},"page":"24237-24248","source":"Crossref","is-referenced-by-count":1,"title":["Azoreductase-Responsive\nNIR Fluorescent Self-Assembled\nProbe for Hypoxia Imaging of Tumors, Hindlimb Ischemia, and Ulcerative\nColitis"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1572-3362","authenticated-orcid":true,"given":"Li-Song","family":"Zhang","sequence":"first","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian-Hang","family":"Wang","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shi-Ting","family":"Weng","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai-Yan","family":"Cui","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen-Wen","family":"Zhao","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chun","family":"Liu","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing-Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"Tianjin Institute for Drug Control (TIDC) , ,","place":["Tianjin, China, 300070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu-Yuan","family":"Li","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5988-1557","authenticated-orcid":true,"given":"Zhi-Song","family":"Zhang","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300350"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"issue":"6","key":"2026083118460419500_cit1","doi-asserted-by":"publisher","first-page":"1075","DOI":"10.1016\/j.immuni.2020.05.001","article-title":"Single-Cell RNA Sequencing of Tumor-Infiltrating NK\nCells Reveals that Inhibition of Transcription Factor HIF-1\u03b1\nUnleashes NK Cell Activity","volume":"52","author":"Ni","year":"2020","journal-title":"Immunity"},{"key":"2026083118460419500_cit2","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2022.123834","article-title":"A highly sensitive NIR fluorescence\nprobe for hypoxia\nimaging in cells and ulcerative colitis","volume":"252","author":"Wang","year":"2023","journal-title":"Talanta"},{"issue":"16","key":"2026083118460419500_cit3","doi-asserted-by":"publisher","first-page":"10901","DOI":"10.1021\/acs.analchem.9b02857","article-title":"In-Situ\nImaging of Azoreductase Activity in the Acute and Chronic Ulcerative\nColitis Mice by a Near-Infrared Fluorescent Probe","volume":"91","author":"Tian","year":"2019","journal-title":"Anal. 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