{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T20:13:20Z","timestamp":1783196000525,"version":"3.54.6"},"reference-count":54,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,6]],"date-time":"2022-07-06T00:00:00Z","timestamp":1657065600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["39404578"],"award-info":[{"award-number":["39404578"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["394046768"],"award-info":[{"award-number":["394046768"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["395357507"],"award-info":[{"award-number":["395357507"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["360372040"],"award-info":[{"award-number":["360372040"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The investigation of hepatocarcinogenesis is a major field of interest in oncology research and rodent models are commonly used to unravel the pathophysiology of onset and progression of hepatocellular carcinoma. HCC is a highly vascularized tumor and vascular remodeling is one of the hallmarks of tumor progression. To date, only a few detailed data exist about the vasculature and vascular remodeling in rodent models used for hepatocarcinogenesis. In this study, the vasculature of HCC and the preneoplastic foci of alteration (FCA) of different mouse models with varying genetic backgrounds were comprehensively characterized by using immunohistochemistry (CD31, Collagen IV, \u03b1SMA, Desmin and LYVE1) and RNA in situ hybridization (VEGF-A). Computational image analysis was performed to evaluate selected parameters including microvessel density, pericyte coverage, vessel size, intratumoral vessel distribution and architecture using the Aperio ImageScope and Definiens software programs. HCC presented with a significantly lower number of vessels, but larger vessel size and increased coverage, leading to a higher degree of maturation, whereas FCA lesions presented with a higher microvessel density and a higher amount of smaller but more immature vessels. Our results clearly demonstrate that vascular remodeling is present and crucial in early stages of experimental hepatocarcinogenesis. In addition, our detailed characterization provides a strong basis for further angiogenesis studies in these experimental models.<\/jats:p>","DOI":"10.3390\/cells11142129","type":"journal-article","created":{"date-parts":[[2022,7,6]],"date-time":"2022-07-06T21:15:52Z","timestamp":1657142152000},"page":"2129","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Vascular Remodeling Is a Crucial Event in the Early Phase of Hepatocarcinogenesis in Rodent Models for Liver Tumorigenesis"],"prefix":"10.3390","volume":"11","author":[{"given":"Margaret","family":"Tulessin","sequence":"first","affiliation":[{"name":"Institute of Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rim Sabrina Jahan","family":"Sarker","sequence":"additional","affiliation":[{"name":"Institute of Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"},{"name":"Comparative Experimental Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joscha","family":"Griger","sequence":"additional","affiliation":[{"name":"Institute of Molecular Oncology and Functional Genomics, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7157-1201","authenticated-orcid":false,"given":"Thomas","family":"Leibing","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Venereology, and Allergology, University Medical Center and Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cyrill","family":"Geraud","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Venereology, and Allergology, University Medical Center and Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany"},{"name":"Section of Clinical and Molecular Dermatology, University Medical Center and Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany"},{"name":"European Center for Angioscience (ECAS), Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wilko","family":"Weichert","sequence":"additional","affiliation":[{"name":"Institute of Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"},{"name":"Comparative Experimental Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7269-5433","authenticated-orcid":false,"given":"Katja","family":"Steiger","sequence":"additional","affiliation":[{"name":"Institute of Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"},{"name":"Comparative Experimental Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3400-7254","authenticated-orcid":false,"given":"Carolin","family":"Mogler","sequence":"additional","affiliation":[{"name":"Institute of Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"},{"name":"Comparative Experimental Pathology, School of Medicine, Technical University of Munich (TUM), 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1002\/ijc.33588","article-title":"Cancer statistics for the year 2020: An overview","volume":"149","author":"Ferlay","year":"2021","journal-title":"Int. J. Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1002\/ijc.31937","article-title":"Estimating the global cancer incidence and mortality in 2018: GLOBOCAN sources and methods","volume":"144","author":"Ferlay","year":"2019","journal-title":"Int. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21590","article-title":"Cancer statistics, 2020","volume":"70","author":"Siegel","year":"2020","journal-title":"CA Cancer J. Clin."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","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_5","doi-asserted-by":"crossref","first-page":"9","DOI":"10.4103\/ijmr.IJMR_1456_17","article-title":"Non-alcoholic fatty liver disease associated with hepatocellular carcinoma: An increasing concern","volume":"149","author":"Dhamija","year":"2019","journal-title":"Indian J. Med. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s12328-014-0548-5","article-title":"Clinical features of hepatocellular carcinoma associated with nonalcoholic fatty liver disease: A review of human studies","volume":"8","author":"Oda","year":"2015","journal-title":"Clin. J. Gastroenterol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1634\/theoncologist.2010-S4-14","article-title":"The etiology of hepatocellular carcinoma and consequences for treatment","volume":"15","author":"Sanyal","year":"2010","journal-title":"Oncologist"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"100305","DOI":"10.1016\/j.jhepr.2021.100305","article-title":"Epidemiology of non-alcoholic fatty liver disease and hepatocellular carcinoma","volume":"3","author":"Younossi","year":"2021","journal-title":"JHEP Rep"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"S361","DOI":"10.21037\/jgo.2020.02.08","article-title":"Addressing the worldwide hepatocellular carcinoma: Epidemiology, prevention and management","volume":"12","author":"Samant","year":"2021","journal-title":"J. Gastrointest. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Furuta, K., Guo, Q., Hirsova, P., and Ibrahim, S.H. (2020). Emerging Roles of Liver Sinusoidal Endothelial Cells in Nonalcoholic Steatohepatitis. Biology, 9.","DOI":"10.3390\/biology9110395"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1097\/MCG.0000000000000116","article-title":"World Gastroenterology Organisation global guidelines: Nonalcoholic fatty liver disease and nonalcoholic steatohepatitis","volume":"48","author":"LaBrecque","year":"2014","journal-title":"J. Clin. Gastroenterol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"S5","DOI":"10.1186\/1746-1596-6-S1-S5","article-title":"Digital pathology evaluation of complement C4d component deposition in the kidney allograft biopsies is a useful tool to improve reproducibility of the scoring","volume":"6","author":"Brazdziute","year":"2011","journal-title":"Diagn. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1002\/jgh3.12070","article-title":"Epidemiology of non-alcoholic fatty liver disease-related hepatocellular carcinoma and its implications","volume":"2","author":"Wong","year":"2018","journal-title":"JGH Open"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e98","DOI":"10.1038\/ctg.2017.28","article-title":"Targeting the vasculature in hepatocellular carcinoma treatment: Starving versus normalizing blood supply","volume":"8","author":"Liu","year":"2017","journal-title":"Clin. Transl. Gastroenterol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4157","DOI":"10.1158\/0008-5472.CAN-13-2311","article-title":"An inducible hepatocellular carcinoma model for preclinical evaluation of antiangiogenic therapy in adult mice","volume":"74","author":"Runge","year":"2014","journal-title":"Cancer Res"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"584250","DOI":"10.3389\/fmed.2020.584250","article-title":"Evaluation of Intra-Tumoral Vascularization in Hepatocellular Carcinomas","volume":"7","author":"Zhang","year":"2020","journal-title":"Front. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"S135","DOI":"10.1067\/msy.2002.119365","article-title":"The mechanisms of angiogenesis in hepatocellular carcinoma: Angiogenic switch during tumor progression","volume":"131","author":"Sugimachi","year":"2002","journal-title":"Surgery"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.4161\/cc.5.16.3018","article-title":"Role of angiogenesis in human tumor dormancy: Animal models of the angiogenic switch","volume":"5","author":"Naumov","year":"2006","journal-title":"Cell Cycle"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"12983","DOI":"10.18632\/oncotarget.14677","article-title":"Prolyl hydroxylase domain protein 3 and asparaginyl hydroxylase factor inhibiting HIF-1 levels are predictive of tumoral behavior and prognosis in hepatocellular carcinoma","volume":"8","author":"Ma","year":"2017","journal-title":"Oncotarget"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1111\/j.1478-3231.2006.01243.x","article-title":"Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma: Importance of angiopoietin-2 and hypoxia-induced factor-1 alpha","volume":"26","author":"Wada","year":"2006","journal-title":"Liver Int."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/s10456-021-09780-y","article-title":"Angiodiversity and organotypic functions of sinusoidal endothelial cells","volume":"24","author":"Koch","year":"2021","journal-title":"Angiogenesis"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/nature17040","article-title":"Angiocrine functions of organ-specific endothelial cells","volume":"529","author":"Rafii","year":"2016","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1002\/1097-0142(19900501)65:9<1985::AID-CNCR2820650918>3.0.CO;2-C","article-title":"An immunohistochemical and ultrastructural study of the sinusoids of hepatocellular carcinoma","volume":"65","author":"Haratake","year":"1990","journal-title":"Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.dld.2012.10.008","article-title":"Liver carcinogenesis: Rodent models of hepatocarcinoma and cholangiocarcinoma","volume":"45","author":"Kisseleva","year":"2013","journal-title":"Dig. Liver Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1356","DOI":"10.1007\/s12029-021-00732-9","article-title":"Evaluation of ATAD2 as a Potential Target in Hepatocellular Carcinoma","volume":"52","author":"Ekin","year":"2021","journal-title":"J. Gastrointest. Cancer"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Masuzaki, R., Kanda, T., Sasaki, R., Matsumoto, N., Nirei, K., Ogawa, M., Karp, S.J., Moriyama, M., and Kogure, H. (2022). Suppressors of Cytokine Signaling and Hepatocellular Carcinoma. Cancers, 14.","DOI":"10.3390\/cancers14102549"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"23306","DOI":"10.18632\/oncotarget.4202","article-title":"Mouse models of liver cancer: Progress and recommendations","volume":"6","author":"He","year":"2015","journal-title":"Oncotarget"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Steiger, K., Gross, N., Widholz, S.A., Rad, R., Weichert, W., and Mogler, C. (2020). Genetically Engineered Mouse Models of Liver Tumorigenesis Reveal a Wide Histological Spectrum of Neoplastic and Non-Neoplastic Liver Lesions. Cancers, 12.","DOI":"10.3390\/cancers12082265"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"5S","DOI":"10.1177\/0192623310386499","article-title":"Proliferative and nonproliferative lesions of the rat and mouse hepatobiliary system","volume":"38","author":"Thoolen","year":"2010","journal-title":"Toxicol. Pathol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1293\/tox.25.189","article-title":"Comparative histomorphological review of rat and human hepatocellular proliferative lesions","volume":"25","author":"Thoolen","year":"2012","journal-title":"J. Toxicol. Pathol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1111\/j.1349-7006.1996.tb00244.x","article-title":"Sinusoidal capillarization and arterial blood supply continuously proceed with the advance of the stages of hepatocarcinogenesis in the rat","volume":"87","author":"Yamamoto","year":"1996","journal-title":"Jpn. J. Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1089\/caff.2018.0019","article-title":"Adenosine Kinase Deficiency Increases Susceptibility to a Carcinogen","volume":"9","author":"Owen","year":"2019","journal-title":"J. Caffeine Adenosine Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"875324","DOI":"10.3389\/fmolb.2022.875324","article-title":"Hepatocyte-Specific Knock-Out of Nfib Aggravates Hepatocellular Tumorigenesis via Enhancing Urea Cycle","volume":"9","author":"Zhou","year":"2022","journal-title":"Front. Mol. Biosci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1002\/hep.510300507","article-title":"The vascular profile of regenerative and dysplastic nodules of the cirrhotic liver: Implications for diagnosis and classification","volume":"30","author":"Roncalli","year":"1999","journal-title":"Hepatology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1111\/j.1440-1827.1994.tb02925.x","article-title":"Sinusoidal capillarization in small hepatocellular carcinoma","volume":"44","author":"Kin","year":"1994","journal-title":"Pathol. Int."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.1111\/liv.12262","article-title":"Endothelial transdifferentiation in hepatocellular carcinoma: Loss of Stabilin-2 expression in peri-tumourous liver correlates with increased survival","volume":"33","author":"Geraud","year":"2013","journal-title":"Liver Int."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/s13550-021-00826-2","article-title":"Correlation of in vivo imaging to morphomolecular pathology in translational research: Challenge accepted","volume":"11","author":"Ballke","year":"2021","journal-title":"EJNMMI Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1007\/s00292-018-0458-2","article-title":"Histopathological research laboratories in translational research: Conception and integration into the infrastructure of pathological institutes","volume":"40","author":"Steiger","year":"2019","journal-title":"Pathologe"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"331","DOI":"10.7150\/ntno.39514","article-title":"Microbubble-enhanced ultrasound for the antivascular treatment and monitoring of hepatocellular carcinoma","volume":"3","author":"Sultan","year":"2019","journal-title":"Nanotheranostics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2227","DOI":"10.1007\/s00432-019-02977-1","article-title":"Loss of enhancer of zeste homologue 2 (EZH2) at tumor invasion front is correlated with higher aggressiveness in colorectal cancer cells","volume":"145","author":"Muenzner","year":"2019","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_41","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_42","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1293\/tox.22.11","article-title":"Biological Basis of Differential Susceptibility to Hepatocarcinogenesis among Mouse Strains","volume":"22","author":"Maronpot","year":"2009","journal-title":"J. Toxicol. Pathol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Blidisel, A., Marcovici, I., Coricovac, D., Hut, F., Dehelean, C.A., and Cretu, O.M. (2021). Experimental Models of Hepatocellular Carcinoma\u2014A Preclinical Perspective. Cancers, 13.","DOI":"10.3390\/cancers13153651"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1111\/hepr.12487","article-title":"Microvessel density in hepatocellular carcinoma: Prognostic significance and review of the previous published work","volume":"45","author":"Murakami","year":"2015","journal-title":"Hepatol. Res."},{"key":"ref_45","first-page":"93","article-title":"Angiogenic switch as a molecular target of malignant tumors","volume":"38","author":"Tanaka","year":"2003","journal-title":"J. Gastroenterol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1111\/his.13975","article-title":"The 2019 WHO classification of tumours of the digestive system","volume":"76","author":"Nagtegaal","year":"2020","journal-title":"Histopathology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"903740","DOI":"10.3389\/fcell.2022.903740","article-title":"Mouse Models of Liver Parenchyma Injuries and Regeneration","volume":"10","author":"Du","year":"2022","journal-title":"Front. Cell. Dev. Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1038\/nature12681","article-title":"Divergent angiocrine signals from vascular niche balance liver regeneration and fibrosis","volume":"505","author":"Ding","year":"2014","journal-title":"Nature"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1038\/nature09493","article-title":"Inductive angiocrine signals from sinusoidal endothelium are required for liver regeneration","volume":"468","author":"Ding","year":"2010","journal-title":"Nature"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1126\/science.1244880","article-title":"Endothelial cell-derived angiopoietin-2 controls liver regeneration as a spatiotemporal rheostat","volume":"343","author":"Hu","year":"2014","journal-title":"Science"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"332","DOI":"10.15252\/emmm.201404246","article-title":"Hepatic stellate cell-expressed endosialin balances fibrogenesis and hepatocyte proliferation during liver damage","volume":"7","author":"Mogler","year":"2015","journal-title":"EMBO Mol. Med."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1130","DOI":"10.1038\/labinvest.2015.95","article-title":"Pivotal role of liver sinusoidal endothelial cells in NAFLD\/NASH progression","volume":"95","author":"Miyao","year":"2015","journal-title":"Lab. Investig."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1158\/1541-7786.MCR-07-0172","article-title":"Molecular mechanisms of liver carcinogenesis in the mdr2-knockout mice","volume":"5","author":"Katzenellenbogen","year":"2007","journal-title":"Mol. Cancer. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4001","DOI":"10.1158\/0008-5472.CAN-05-2937","article-title":"Multiple adaptive mechanisms to chronic liver disease revealed at early stages of liver carcinogenesis in the Mdr2-knockout mice","volume":"66","author":"Katzenellenbogen","year":"2006","journal-title":"Cancer Res."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/14\/2129\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:43:37Z","timestamp":1760139817000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/14\/2129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,6]]},"references-count":54,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["cells11142129"],"URL":"https:\/\/doi.org\/10.3390\/cells11142129","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,6]]}}}