Feasibility Study Of Multiphoton Optical Biopsy For Lymph Nodes Around The Root Of Inferior Mesenteric Artery In Rectal Cancer

ObjectiveThe aim of this study is to localize,qualitatively and quantitatively diagnose lymph nodes around the root of inferior mesenteric artery(IMA)(station 253 node)metastasis in rectal cancer by using technology of carbon nanoparticles tracer and multiphoton optical biopsy,and analyze the metastatic regularity of station 253 nodes for further achieve precise navigation and minimally invasive treatment in radical resection of rectal cancerMethodsThis study was divided into three main experimental chapters.Chapter 1:A retrospective study of 502 patients with rectal cancer who underwent radical resection in Nanfang Hospital from August 2014 to August 2018 was performed,they were divided into station 253 nodes positive group(253+)and 253 negative group(253-).Univariate and multivariate logistic regression analysis was used to analyze the clinicopathological data of the two groups,and then established prediction model of 253 lymph node metastasis.Chapter 2:Sixty-six patients were enrolled in this study.Patients were divided into carbon nanoparticle(CN)group and control(CL)group and matched with 1:1 manner based on age(± 3),gender,BMI(± 2),tumor location,pre-operative tumor stage,and those with or without neoadjuvant therapy.Thirty-three patients from CN group were endoscopically injected carbon nanoparticles into primary tumor one day before surgery.All patients were subjected to laparoscopic radical resection with D3 lymph node dissection.Intra-operative and post-operative data were recorded and compared between two groups.Chapter 3:253 lymph nodes from 60 patients with rectal cancer were selected in this study,these then underwent multiphoton imaging examination.The redox rate FAD/NADH of normal and tumors cells were measured by using multiphoton imaging to further reveal cell metabolism.Then,the diagnostic characteristics of metastatic lymph nodes were established via multiphoton imaging compared with the final HE pathology,and then a blinded study was performed to test the sensitivity,specificity,and accuracy of multiphoton imaging by investigating 48 patients with rectal cancer.Results1.40 patients had 253 positive lymph nodes(253+)and 462 patients had 253 negative lymph nodes(253-).Multivariate logistic regression analysis results showed that less than 60 years old,female,poor differentiation,T3-4 and Mi were independent risk factors for station 253 nodes metastasis.2.Black-dyed station 253 nodes were easily found under laparoscopy.There was no significant difference in intraoperative and postoperative recovery between the two groups.Significantly higher number of station 251 nodes,253 nodes and total lymph nodes were harvested in CN group compared to CL group.3.Multiphoton imaging showed the size and arrangement of lymphocyte were irregular in metastatic lymph nodes.There was a small amount of irregular collagen structure or elastic fibrous tissue around lymphocyte.Cancer cells display irregular size and shape,cellular and nuclear pleomorphism,enlarged nuclei,increased nuclear-cytoplasmic ratio.The redox rate FAD/NADH in tumor cells was also significantly higher than that in normal lymphocytes.Compared with the final HE pathological diagnosis,the sensitivity,specificity and accuracy of multiphoton imaging were 83.3%,97.6%and 95.8%,respectively.ConclusionPreoperative localization using carbon nanoparticle tracer technique,intraoperative qualitative and quantitative diagnose using multiphoton optical biopsy technique to evaluate whether station 253 node have metastasis in rectal cancer,which was helpful to determine the level of IMA ligation and 253 lymph nodes dissection in individual rectal cancer patients.