{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T05:06:52Z","timestamp":1790140012625,"version":"4.0.1"},"reference-count":66,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82274593"],"award-info":[{"award-number":["82274593"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["82274593"],"award-info":[{"award-number":["82274593"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Natural Science Foundation of Hebei Province","award":["H2023206137"],"award-info":[{"award-number":["H2023206137"]}]},{"DOI":"10.13039\/501100008238","name":"S&T Program of Hebei","doi-asserted-by":"publisher","award":["223777122D"],"award-info":[{"award-number":["223777122D"]}],"id":[{"id":"10.13039\/501100008238","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008238","name":"Hebei Province Young Top Talents Programme","doi-asserted-by":"publisher","award":["BJK2024113"],"award-info":[{"award-number":["BJK2024113"]}],"id":[{"id":"10.13039\/501100008238","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Hebei Province Traditional Chinese Medicine Scientific Research Subjects Programme","award":["2024110"],"award-info":[{"award-number":["2024110"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Esophageal squamous cell carcinoma frequently develops through a prolonged precancerous phase, creating an opportunity for intervention. Zishui Qinggan Yin (ZSQGY) is a multi-component botanical preparation, but its effects in esophageal precancer and its formulation-level chemical profile remain incompletely defined. We monitored a predefined 40-analyte panel by UPLC-Q Exactive\/MS in three independently extracted aliquots from the same formulation batch administered to mice. A 4-nitroquinoline-1-oxide model was then evaluated by consensus histopathology, mouse-grouped whole-slide digital pathology, network-based target prioritization, molecular docking, exploratory drug affinity responsive target stability (DARTS), Western blotting, qPCR, and serial-section immunohistochemistry. Signals corresponding to 38 predefined analytes were reported. ZSQGY treatment was associated with a lower-grade lesion pattern in the evaluated mouse-level specimens. Exploratory DARTS was analyzed in two independent n = 6 tissue-background datasets. In the original 4-NQO model-background cohort, ex vivo ZSQGY (100 \u03bcg\/mL) increased the AKT1-immunoreactive relative-protection signal in five of six lysates (mean 2.04 \u00b1 1.15). In an additional cohort from mice that had received ZSQGY in vivo, ex vivo ZSQGY produced mean relative-protection values of 1.21, 1.75, and 1.92 at 50, 100, and 200 \u03bcg\/mL, respectively. The nominal exact p value for the 100 \u03bcg\/mL versus vehicle contrast in the additional exploratory dataset was 0.0313; This exploratory result should be interpreted cautiously. Neither dataset establishes direct binding or in vivo target occupancy. PI3K p110\u03b1 DARTS was technically inconclusive. Endpoint tissues showed lower AKT, mTOR, S6K1, and 4E-BP1 phosphorylation, together with PTEN-associated changes, reduced Ki-67, increased cleaved caspase-3 staining, and selected downstream transcript changes, including a non-significant downward trend in PARP1. These findings support a preclinical association between ZSQGY treatment, attenuation of esophageal precancer progression, and AKT-centered PI3K\/AKT\/mTOR pathway modulation. They do not establish direct binding, in vivo target occupancy, pathway necessity, active constituents, or causal mediation.<\/jats:p>","DOI":"10.3390\/life16091574","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T12:50:03Z","timestamp":1789995003000},"page":"1574","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeted Chemical Monitoring and Integrated Preclinical Evaluation of Zishui Qinggan Yin in 4-NQO-Induced Esophageal Precancer: Histopathological and PI3K\/AKT\/mTOR-Associated Changes"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-7971-8821","authenticated-orcid":false,"given":"Yueheng","family":"Ouyang","sequence":"first","affiliation":[{"name":"College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-7599-4149","authenticated-orcid":false,"given":"Jianxin","family":"Guo","sequence":"additional","affiliation":[{"name":"College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Jianfeng","family":"Yuan","sequence":"additional","affiliation":[{"name":"College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Zhongbing","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"project-administration","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0592-2712","authenticated-orcid":false,"given":"Jing","family":"Li","sequence":"additional","affiliation":[{"name":"College of Integrated Chinese and Western Medicine, Hebei Medical University, Shijiazhuang 050017, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"ref_1","first-page":"209","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.1002\/cam4.4965","article-title":"Progression of precancerous lesions of esophageal squamous cell carcinomas in a high-risk, rural Chinese population","volume":"12","author":"Gao","year":"2023","journal-title":"Cancer Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1038\/s41568-024-00754-y","article-title":"Emerging strategies to investigate the biology of early cancer","volume":"24","author":"Zhou","year":"2024","journal-title":"Nat. Rev. Cancer"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1158\/2159-8290.CD-24-0015","article-title":"Advancing Cancer Interception","volume":"14","author":"Domchek","year":"2024","journal-title":"Cancer Discov."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s41021-021-00180-8","article-title":"Cancer interception by interceptor molecules: Mechanistic, preclinical and human translational studies with chlorophylls","volume":"43","author":"Dashwood","year":"2021","journal-title":"Genes Environ."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"52","DOI":"10.4103\/jcrt.JCRT_805_17","article-title":"Clinical efficacy of endoscopic submucosal dissection in the treatment of early esophageal cancer and precancerous lesions","volume":"14","author":"Wu","year":"2018","journal-title":"J. Cancer Res. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2038","DOI":"10.1016\/j.ccell.2023.11.003","article-title":"Genomic alterations driving precancerous to cancerous lesions in esophageal cancer development","volume":"41","author":"Chang","year":"2023","journal-title":"Cancer Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"50071","DOI":"10.18632\/oncotarget.18412","article-title":"CDKN2A methylation in esophageal cancer: A meta-analysis","volume":"8","author":"Zhou","year":"2017","journal-title":"Oncotarget"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.cell.2017.04.001","article-title":"AKT\/PKB signaling: Navigating the network","volume":"169","author":"Manning","year":"2017","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1016\/j.cell.2017.02.004","article-title":"mTOR signaling in growth, metabolism, and disease","volume":"168","author":"Saxton","year":"2017","journal-title":"Cell"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Luo, Q., Du, R., Liu, W., Huang, G., Dong, Z., and Li, X. (2022). PI3K\/Akt\/mTOR signaling pathway: Role in esophageal squamous cell carcinoma, regulatory mechanisms and opportunities for targeted therapy. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.852383"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Huang, R., Dai, Q., Yang, R., Duan, Y., Zhao, Q., Haybaeck, J., and Yang, Z. (2022). A review: PI3K\/AKT\/mTOR signaling pathway and its regulated eukaryotic translation initiation factors may be a potential therapeutic target in esophageal squamous cell carcinoma. Front. Oncol., 12.","DOI":"10.3389\/fonc.2022.817916"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1186\/s12935-019-0998-4","article-title":"TRIM27 promotes the development of esophagus cancer via regulating PTEN\/AKT signaling pathway","volume":"19","author":"Ma","year":"2019","journal-title":"Cancer Cell Int."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.humpath.2012.05.011","article-title":"Clinicopathologic significance and function of mammalian target of rapamycin activation in esophageal squamous cell carcinoma","volume":"44","author":"Kim","year":"2013","journal-title":"Hum. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1186\/s13046-015-0212-z","article-title":"Phosphorylated AKT1 is associated with poor prognosis in esophageal squamous cell carcinoma","volume":"34","author":"Zhu","year":"2015","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1038\/s41591-019-0508-1","article-title":"Clinical-grade computational pathology using weakly supervised deep learning on whole-slide images","volume":"25","author":"Campanella","year":"2019","journal-title":"Nat. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1038\/s41551-020-00682-w","article-title":"Data-efficient and weakly supervised computational pathology on whole-slide images","volume":"5","author":"Lu","year":"2021","journal-title":"Nat. Biomed. Eng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6572","DOI":"10.1038\/s41467-022-34025-x","article-title":"Uncertainty-informed deep learning models enable high-confidence predictions for digital histopathology","volume":"13","author":"Dolezal","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1177\/03009858231151381","article-title":"A method for scoring 4-nitroquinoline 1-oxide-induced murine esophageal squamous neoplasia","volume":"60","author":"Aziz","year":"2023","journal-title":"Vet. Pathol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Liu, Z., Su, R., Ahsan, A., Liu, C., Liao, X., Tian, D., and Su, M. (2022). Esophageal squamous cancer from 4NQO-induced mice model: CNV alterations. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232214304"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"117006","DOI":"10.1016\/j.taap.2024.117006","article-title":"Ethanol exposure exacerbates 4-nitroquinoline-1-oxide induced esophageal carcinogenesis and induces invasive carcinoma with muscularis propria infiltration in a mouse model","volume":"489","author":"Huang","year":"2024","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Percie du Sert, N., Hurst, V., Ahluwalia, A., Alam, S., Avey, M.T., Baker, M., Browne, W.J., Clark, A., Cuthill, I.C., and Dirnagl, U. (2020). The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol., 18.","DOI":"10.1371\/journal.pbio.3000410"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"16878","DOI":"10.1038\/s41598-017-17204-5","article-title":"QuPath: Open-source software for digital pathology image analysis","volume":"7","author":"Bankhead","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Macenko, M., Niethammer, M., Marron, J.S., Borland, D., Woosley, J.T., Guan, X., Schmitt, C., and Thomas, N.E. (July, January 28). A method for normalizing histology slides for quantitative analysis. Proceedings of the 2009 IEEE International Symposium on Biomedical Imaging, Boston, MA, USA.","DOI":"10.1109\/ISBI.2009.5193250"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Selvaraju, R.R., Cogswell, M., Das, A., Vedantam, R., Parikh, D., and Batra, D. (2017, January 22\u201329). Grad-CAM: Visual explanations from deep networks via gradient-based localization. Proceedings of the 2017 IEEE International Conference on Computer Vision (ICCV), Venice, Italy.","DOI":"10.1109\/ICCV.2017.74"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"D1373","DOI":"10.1093\/nar\/gkac956","article-title":"PubChem 2023 update","volume":"51","author":"Kim","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"W357","DOI":"10.1093\/nar\/gkz382","article-title":"SwissTargetPrediction: Updated data and new features for efficient prediction of protein targets of small molecules","volume":"47","author":"Daina","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"The UniProt Consortium (2023). UniProt: The Universal Protein Knowledgebase in 2023. Nucleic Acids Res., 51, D523\u2013D531.","DOI":"10.1093\/nar\/gkac1052"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Safran, M., Rosen, N., Twik, M., BarShir, R., Stein, T.I., Dahary, D., Fishilevich, S., and Lancet, D. (2022). The GeneCards Suite. Practical Guide to Life Science Databases, Springer.","DOI":"10.1007\/978-981-16-5812-9_2"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"D36","DOI":"10.1093\/nar\/gku1055","article-title":"Gene: A gene-centered information resource at NCBI","volume":"43","author":"Brown","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"D1257","DOI":"10.1093\/nar\/gkac833","article-title":"Comparative Toxicogenomics Database (CTD): Update 2023","volume":"51","author":"Davis","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"D605","DOI":"10.1093\/nar\/gkaa1074","article-title":"The STRING database in 2021: Customizable protein-protein networks, and functional characterization of user-uploaded gene\/measurement sets","volume":"49","author":"Szklarczyk","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"The Cancer Genome Atlas Research Network (2017). Integrated genomic characterization of oesophageal carcinoma. Nature, 541, 169\u2013175.","DOI":"10.1038\/nature20805"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e47","DOI":"10.1093\/nar\/gkv007","article-title":"limma powers differential expression analyses for RNA-sequencing and microarray studies","volume":"43","author":"Ritchie","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: A software environment for integrated models of biomolecular interaction networks","volume":"13","author":"Shannon","year":"2003","journal-title":"Genome Res."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Bader, G.D., and Hogue, C.W.V. (2003). An automated method for finding molecular complexes in large protein interaction networks. BMC Bioinform., 4.","DOI":"10.1186\/1471-2105-4-2"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"The Gene Ontology Consortium (2019). The Gene Ontology Resource: 20 years and still GOing strong. Nucleic Acids Res., 47, D330\u2013D338.","DOI":"10.1093\/nar\/gky1055"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"D545","DOI":"10.1093\/nar\/gkaa970","article-title":"KEGG: Integrating viruses and cellular organisms","volume":"49","author":"Kanehisa","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"D587","DOI":"10.1093\/nar\/gkac963","article-title":"KEGG for taxonomy-based analysis of pathways and genomes","volume":"51","author":"Kanehisa","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS J. Integr. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1038\/nchembio.118","article-title":"Network pharmacology: The next paradigm in drug discovery","volume":"4","author":"Hopkins","year":"2008","journal-title":"Nat. Chem. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13020-024-01056-z","article-title":"Network pharmacology: A crucial approach in traditional Chinese medicine research","volume":"20","author":"Zhai","year":"2025","journal-title":"Chin. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"21984","DOI":"10.1073\/pnas.0910040106","article-title":"Target identification using drug affinity responsive target stability (DARTS)","volume":"106","author":"Lomenick","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","first-page":"4244","article-title":"Use of a monoclonal anti-estrogen receptor antibody in the immunohistochemical evaluation of human tumors","volume":"46","author":"McCarty","year":"1986","journal-title":"Cancer Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_48","first-page":"65","article-title":"A simple sequentially rejective multiple test procedure","volume":"6","author":"Holm","year":"1979","journal-title":"Scand. J. Stat."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2852","DOI":"10.1101\/gad.912401","article-title":"Hierarchical phosphorylation of the translation inhibitor 4E-BP1","volume":"15","author":"Gingras","year":"2001","journal-title":"Genes Dev."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"13375","DOI":"10.1074\/jbc.273.22.13375","article-title":"The tumor suppressor, PTEN\/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate","volume":"273","author":"Maehama","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1002\/(SICI)1097-4652(200003)182:3<311::AID-JCP1>3.0.CO;2-9","article-title":"The Ki-67 protein: From the known and the unknown","volume":"182","author":"Scholzen","year":"2000","journal-title":"J. Cell. Physiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1111\/j.1745-7270.2005.00108.x","article-title":"Caspase family proteases and apoptosis","volume":"37","author":"Fan","year":"2005","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1038\/sj.cdd.4400476","article-title":"Emerging roles of caspase-3 in apoptosis","volume":"6","author":"Porter","year":"1999","journal-title":"Cell Death Differ."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1111\/j.1442-2050.2005.00463.x","article-title":"Cyclin D1, E2F1 expression levels are associated with characteristics and prognosis of esophageal squamous cell carcinoma","volume":"18","author":"Mega","year":"2005","journal-title":"Dis. Esophagus"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1038\/nrc3090","article-title":"Cyclin D as a therapeutic target in cancer","volume":"11","author":"Musgrove","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1016\/S1097-2765(05)00089-4","article-title":"Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin, LY294002, quercetin, myricetin and staurosporine","volume":"6","author":"Walker","year":"2000","journal-title":"Mol. Cell"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Wu, W.-I., Voegtli, W.C., Sturgis, H.L., Dizon, F.P., Vigers, G.P.A., and Brandhuber, B.J. (2010). Crystal structure of human AKT1 with an allosteric inhibitor reveals a new mode of kinase inhibition. PLoS ONE, 5.","DOI":"10.2210\/pdb3o96\/pdb"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1126\/science.1106148","article-title":"Phosphorylation and regulation of Akt\/PKB by the rictor-mTOR complex","volume":"307","author":"Sarbassov","year":"2005","journal-title":"Science"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"132531","DOI":"10.1016\/j.foodchem.2022.132531","article-title":"Plant flavonoids: Classification, distribution, biosynthesis, and antioxidant activity","volume":"383","author":"Shen","year":"2022","journal-title":"Food Chem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"e47","DOI":"10.1017\/jns.2016.41","article-title":"Flavonoids: An overview","volume":"5","author":"Panche","year":"2016","journal-title":"J. Nutr. Sci."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Zughaibi, T.A., Suhail, M., Tarique, M., and Tabrez, S. (2021). Targeting PI3K\/Akt\/mTOR Pathway by Different Flavonoids: A Cancer Chemopreventive Approach. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222212455"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1126\/science.1233606","article-title":"Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay","volume":"341","author":"Jafari","year":"2013","journal-title":"Science"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1007\/s13238-013-2084-3","article-title":"NF-\u03baB and STAT3 signaling pathways collaboratively link inflammation to cancer","volume":"4","author":"Fan","year":"2013","journal-title":"Protein Cell"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1126\/science.aaf2403","article-title":"Visualization and analysis of gene expression in tissue sections by spatial transcriptomics","volume":"353","author":"Vickovic","year":"2016","journal-title":"Science"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1016\/j.cell.2018.07.010","article-title":"Deep profiling of mouse splenic architecture with CODEX multiplexed imaging","volume":"174","author":"Goltsev","year":"2018","journal-title":"Cell"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/s41392-023-01710-2","article-title":"Aberrant epithelial cell interaction promotes esophageal squamous-cell carcinoma development and progression","volume":"8","author":"Chen","year":"2023","journal-title":"Signal Transduct. Target. Ther."}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/9\/1574\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T04:40:34Z","timestamp":1790138434000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/9\/1574"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,21]]},"references-count":66,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["life16091574"],"URL":"https:\/\/doi.org\/10.3390\/life16091574","relation":{},"ISSN":["2075-1729"],"issn-type":[{"value":"2075-1729","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,21]]}}}