Background and purpose: When patients have positive sentinel lymph node (SLN), axillary lymph node dissection (ALND) is usually performed, but most of them have no metastasis in the non-sentinel lymph node (nSLN). It is of great significance to predict metastasis of nSLN precisely. The aim of the study was to establish a nomogram for the intraoperative prediction of nSLN metastasis in breast cancer patients using one-step nucleic acid amplification (OSNA) techniques and to direct the subsequent therapy for breast cancer effectively. Methods: Of 552 breast cancer patients who underwent SLN biopsy in the 2010 OSNA clinical trial, 103 with SLN metastasis treated with ALND were assessed to establish a nomogram for intraoperative prediction of nSLN based on the molecular diagnosis. A validation cohort of 61 patients who met the similar criteria in the 2015 OSNA clinical trial subsequently validated it. Results: Primary tumor size, total tumor load, the number of positive SLNs and negative SLNs were associated with the presence of nSLN metastasis based on the multivariable logistic regression results, and a nomogram was established with these variables. Its area under the ROC curve was 0.814 for the predictive model and it was 0.842 in the re-validation cohort. The tumor size assessed by the postoperative histological examination was replaced by the size evaluated by the imaging examination, and the area under the ROC curve was 0.838. There was no statistically significant difference in the accuracy compared with the former validation data (P=0.7406). Conclusion: The predictive nomogram based on the molecular diagnosis can predict the nSLN metastases intra/post-operatively. It appears to be obviously superior to other predictive models and may help to guide the axillary management and to make decisions about radiation target region.

Background and purpose:Sentinel lymph node biopsy has replaced axillary lymph node dissec-tion as the standard staging procedure in early breast cancer patients with clinically negative axillary lymph nodes. It is a critical step for staging and treatment. This study investigated the localization effect of a novel tracer for breast cancer sentinel lymph node biopsy [indocyanine green (ICG)-rituximab (R)], using the hind limb drainage in mice as an animal model.Methods:For exploring the optimal dose and imaging time, different doses of ICG-R were injected subcutane-ously to the dorsum of the foot in the BALB/c mice. Then the lfuorescence vasculature imaging instrument was used continuously to observe the popliteal fossa lymph node (as sentinel lymph node) from the injection to 3 h after injection. For exploring the sustained localization effect, the optimal dose of ICG-R was injected and the imaging instrument was used from imaging to 24 h after injection.Results:The time from injection to imaging and the time from injection to the optimal imaging were shortened with the increased doses, and the imaging rate of the second or third level node increased accordingly. The best dosage of the novel tracer was 0.12 μg (dosage of indocyanine green) and the time from injection to the optimal imaging was about 34 min. After the observation for 24 h, the imaging rate of sentinel lymphnode was maintained at 100%, and the imaging rate of the second and the third level lymph node increased from 0% to 20% and 10%, respectively.Conclusion:ICG-R could clearly locate the sentinel lymph node. There is no imaging of the second level lymph node within 6 h. The novel tracer has high value in the clinical application.

Background and purpose:Sentinel lymph node biopsy is regarded as the standard of care in pa-tients without clinical axillary lymph node metastases in early-stage breast cancer. Accurate detection of sentinel lymph node is an important step for staging, prognosis, and treatment. In this study, a new sentinel lymph node tracer was produced by the rituximab to combine with the lfuorescence tracer (indocyanine green, ICG), and to identify the most appropriate combination ratio of the two agents. Its biological property and safety limitation were evaluated.Methods:Rituximab was combined directly with ICG. The new tracer was analyzed for labeled rate by instant thin-layer chroma-tography-silica gel, molecular integrity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and molecular immune activity by ELLAS. The safety limitation was tested according to the Chinese Pharmacopeia. The localization ability of sentinel lymph node was tested in mice.Results:The new tracer was intact and kept the immune activity of rituximab. The ICG labeled rate of rituximab was 100%. The new tracer was bacteria and pyogen free, and was safe to body with location injection. The most appropriate combination ratio of rituximab and ICG was 4∶1 and 6∶1 with the best sentinel lymph node imaging. The location of sentinel lymph node identiifed by the new tracer was accorded with the radiotracer.Conclusion:The preparation method of the new sentinel lymph node tracer is simple and no radioactive injury. The new tracer has no bacteria, no pyogen and no acute toxicity, and can be used in sentinel lymph node visual-ization.

PURPOSE: Sentinel lymph node biopsy (SLNB), a critical staging and treatment step, has replaced axillary lymph node (LN) dissection as the standard staging procedure for early stage breast cancer patients with clinically negative axillary LNs. Hence, using a murine sentinel lymph node (SLN) model, we investigated the localization effect of the new receptor-targeted tracer, indocyanine green (ICG)-rituximab, on breast cancer SLNB. METHODS: After establishing the murine SLN model, different doses of ICG-rituximab were subcutaneously injected into the hind insteps of BALB/c mice to determine the optimal dose and imaging time using continuous (> 3 hours) MDM-I fluorescence vasculature imaging. To explore the capacity of ICG-rituximab for sustained SLN localization with the optimal dose, MDM-I imaging was monitored at 6, 12, and 24 hours. RESULTS: The popliteal LN was defined as the SLN for hindlimb lymphatic drainage, the iliac LN as the secondary, and the para-aortic or renal LN as the tertiary LNs. The SLN initial imaging and optimal imaging times were shortened with increased ICG-rituximab doses, and the imaging rates of the secondary and tertiary LNs increased accordingly. The optimal ICG dose was 0.12 μg, and its optimal imaging time was 34 minutes. After 24 hours, the SLN imaging rate remained 100%, while those of the secondary and the tertiary LNs increased from 0% (6 hours) and 0% (6 hours) to 10% (12 hours) and 10% (12 hours) to 20% (24 hours) and 10% (24 hours), respectively. CONCLUSION: ICG-rituximab localized to the SLN without imaging from the secondary or tertiary LNs within 6 hours. The optimal ICG dose was 0.12 μg, and the optimal interval for SLN detection was 34 minutes to 6 hours post-injection. This novel receptor-targeted tracer is of great value to clinical research and application.
Animals ; Breast Neoplasms ; Breast ; Drainage ; Fluorescence ; Hindlimb ; Humans ; Indocyanine Green ; Lymph Nodes ; Mice ; Models, Animal ; Rituximab ; Sentinel Lymph Node Biopsy

Objective:To explore the consistency of peripheral whole blood and venous serum procalcitonin (PCT) levels, and the value of peripheral whole blood PCT in evaluating pediatric bacterial infection.Methods:This multicenter cross-sectional parallel control study was conducted in 11 children′s hospital. All the 1 898 patients older than 28 days admitted to these hospitals from March 2018 to February 2019 had their peripheral whole blood and venous serum PCT detected simultaneously with unified equipment, reagent and method. According to the venous serum PCT level, the patients were stratified to subgroups. Analysis of variance and chi-square test were used to compare the demographic characteristics among groups. And the correlation between the peripheral blood and venous serum PCT level was investigated by quantitative Pearson correlation analysis.The PCT resultes were also converted into ranked data to further test the consistency between the two sampling methods by Spearman′s rank correlation test. Furthermore, the ranked data were converted into binary data to evaluate the consistency and investigate the best cut-off of peripheral blood PCT level in predicting bacterial infection.Results:A total of 1 898 valid samples were included (1 098 males, 800 females),age 27.4(12.2,56.7) months. There was a good correlation between PCT values of peripheral whole blood and venous serum ( r=0.97 , P<0.01). The linear regression equation was PCT?venous serum=0.135+0.929×PCT peripheral whole blood. However, when stratified to 5 levels, PCT results showed diverse and unsatisfied consistency between the two sampling methods ( r=0.51-0.92, all P<0.01). But after PCT was converted to ordinal categorical variables, the stratified analysis showed that the coincidence rate of the measured values by the two sampling methods in each boundary area was 84.9%-97.1%. The dichotomous variables also showed a good consistency (coincidence rate 96.8%-99.3%, Youden index 0.82-0.89). According to the severity of disease, the serum PCT value was classified into 4 intervals（<0.5、0.5-<2.0、2.0-<10.0、≥10.0 μg/L）, and the peripheral blood PCT value also showed a good predictive value (AUC value was 0.991 2-0.997 9). The optimal cut points of peripheral whole blood PCT value 0.5、1.0、2.0、10.0 μg/L corresponding to venous serum PCT values were 0.395, 0.595, 1.175 and 3.545 μg/L, respectively. Conclusions:There is a good correlation between peripheral whole blood PCT value and the venous serum PCT value, which means that the peripheral whole blood PCT could facilitate the identification of infection and clinical severity. Besides, the sampling of peripheral whole blood is simple and easy to repeat.