| Objective:The first part of the present study is to evaluate the clinical value of multiple cross-section touch imprint cytology(TIC) as the intraoperative assessment for sentinel lymph nodes(SLN).The second part of the present study is detect the micrometastasis foci in sentinel lymph node from patients with early-stage breast cancer with the varies methods,such as serial section with H&E staining,immunohistochemistry staining,RT-PCR and flow cytometry and to establish a suitable protocol on the pathologic evaluation of sentinel lymph nodes postoperatively in our own institute.Methods:Part one:The Optimization of Intraoperative Touch Imprint Cytology Evaluation of Sentinel Lymph Node for Breast Cancer 366 patients enrolled in the study between Feb-2005 and Feb-2008,whose SLNs captured in the surgery were sliced along the long axis at a 2.0～3.0mm interval. 122 patients participated in the study between Mar-2008 and Jan-2009,whose SLNs harvested were sliced along the short axis at a 1.5mm interval with our own designed cutter.Serial section at a 100μm interval with H&E staining was used as gold standard for pathologic diagnosis.Part two:The Application of Serial Section,IHC,RT-PCR and Flow Cytometry in Detection of Micrometastasis in SLN for Breast Cancer From May-2008 to May-2009,sentinel lymph nodes captured in the surgery,which were sliced with the sentinel lymph node cutter into pieces at a thickness of 1.5mm,were divided into three groups,every third slices were sent for evaluation by serial section with H&E staining and IHC staining,every(3n-1) slices were sent for evaluation by RT-PCR to detect micrometastasis while every (3n-2) sliced were sent for flow cytometry.Results:Part one:Multiple cross-section TIC has a sensitivity,specificity and overall accuracy rate of 92.0%,99.0%and 97.5%,respectively on a per patient basis superior to the standard proposal for preparing imprints.The ability to detect macrometastasis on a per patient base between two proposals had significant difference(p=0.023).The metastasis rates of patients with DCIS, DCIS-Mi and IDC were 4.4%,12.5%and 31.3%,respectively.97.7%of the patients with node involved,whose first involved SLN appeared within the first three SLNs harvested.Part two:Among all the 104 sentinel lymph nodes,serial section with H&E staining discovered 11(10.6%) involved nodes,among which,2 nodes contained micrometastasis.Consequent IHC staining ensured that the 2 nodes mentioned before were macrometastasis and discovered another 11 micrometastasis and 1 macrometastasis nodes among the H&E staining negative nodes.Further more, RT-PCR and flow cytometry discovered another 5(6.2%) and 4(4.9%) nodes, respectively that contained cancer cells.About 40%sentinel lymph nodes micrometastasises detected by IHC staining were accompanied by additional sentinel lymph nodes metastases or micrometastasis(p=0.007).Conclusion:Part one:Multiple cross-section TIC is superior to the standard proposal, especially for the ability to locate macrometastasis.To limit the intraoperative assessment to the first three retrieved nodes may reduce the pathology laboratory and expanse.It could be taken into consideration to spare intraoperative assessment of SLNs for patients with DCIS diagnosed by open biopsy.Part two:The sensitivity of serial section with IHC staining was higher than that of serial section with H&E staining,while RT-PCR and flow cytometry were proved to be more sensitivity.However,no further evidence could proved whether the positive result was a real one.Therefore,serial section with IHC staining was still suggested to be the gold standard for pathologic evaluation of sentinel lymph node postoperatively.The patients with positive nodes only detected by RT-PCR or flow cytometry should be close follow-up. The micrometastasises detected by IHC staining were likely to accompanied by additional sentinel lymph nodes metastases or micrometastasis,therefore,the number of sentinel lymph node captured with the surgery was suggested to be at least two in order to increase the overall accuracy.
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