Background: The regression model of positive nodes in breast cancer after neoadjuvant chemotherapy (NAC) remains controversial. This study aimed to investigate this regression model by injecting and tracing carbon nanoparticles (CNs) into the fusion node prior to NAC in patients with breast cancer.Methods: Guided by ultrasound, 0.3 mL of CNs suspension was injected in a fusion node prior to NAC in 110 patients with local advanced breast cancer. Patients underwent breast surgery and total axillary lymph node dissection following 2-6 cycles of NAC. The distribution by intercostobrachial nerves (ICBN) of positive nodes, black-stained nodes and lymphovascular invasion was investigated by response to NAC.Results: When patients were ranked by response to NAC (from sensitive to resistance), the number of positive nodes increased, as did the proportion of lymphovascular invasion, the number of blackstained nodes decreased. A significantly negative relationship was found between the number of positive nodes and the number of black-stained nodes (p < 0.001). The positive nodes in patients with sensitive consequence followed the rule from under the ICBN to above the ICBN. However, there was counter-example (skip metastasis) in the patients with resistance result.Conclusion: The regression model of positive nodes follows the rule from upper to under, inner to outer in the patients with sensitive consequence to NAC. Long-term staining and tracing by CNs might provide an acceptable and feasible technique to investigate the regression model of positive nodes, and would be a potential method for NAC-treated patients by using of ICBN.Trial registration: NCT 03355261. Retrospectively registered on November 28, 2017. SLNB after NAC as unacceptable when compared with the <5% rate produced by the pioneers of SLNB [3]. In fact, about 9% of patients with positive nodes and T0-2 breast cancer have residual positive nodes in the Level III region after NAC [4]. Therefore, it is essential to explore the regression model post-NAC in patients who are node-positive prior to NAC.Studies from developed countries suggest that SLNB is a reliable method for lymph node evaluation in NAC-treated patients with breast cancer [5, 6]. However, they did not distinguish or exclude patients who were suspected or proven to harbor axillary metastases before receiving NAC. We speculated that the number of patients with N3 nodes in these studies was small. The proportion of patients in China with advanced stage breast cancer is higher than that in more developed countries [7].Therefore, Chinese surgeons are interested in the feasibility of SLNB for patients with local advanced breast cancer who are treated with NAC. Recent studies suggest that SLNB is technically feasible and accurate enough for axillary staging in initially clinically node-negative breast cancer patients after NAC [8]. Additionally, SLNB is feasible for patients whose axillary lymph nodal status is N1 before NAC; however, SLNB cannot be used as a reliable indicator of non-SLN status in N2-3 p...