| Since the overall survival of BCS(breast conserving surgery)and MRM(modified radical mastectomy)is similar in the NSABP B-06 report,BCS has been popularized in the breast cancer patients.Now about 60% patients accept BCS in USA,and about 20% in our country.The percentage of BCS is increasing by year.The aim of BCS is to achieve clear margins to avoid re-excision and achieve good esthetic results.But until now there is no rapid,accurate and ideal enough diagnosis methods to achieve the aim.Raman spectroscopy is a simple,powerful,rapid analytical technology,can provide a quickly intraoperative diagnose of the margin without any pretreatment.So it is a very useful method in clinical.However the composition of the fresh tissues are complicated,and it is very hard to test the aim positions accurately,which leads to the bigger differences of testing results of Raman spectra,the accuracy and specificity of diagnosis model is difficult to meet the needs of the clinical application even using the mathematics methods.We use mapping method of Raman spectroscopy to eliminate the interference of this,and increase the accuracy rate of diagnosis,which is point by point scanning detection in the fixed area.Then the image of the test area was formed by using different methods.It is the base of localized the positive lesions directly.Nano fluorescent material is a kind of tumor cell imaging method,which can quickly display the tumor cell by fluorescence but no other cells,so it could become a new method of cutting edge detection in the future.In addition,Fluorescent biological imaging technology is one of the most important methods for early-stage cancer diagnosis and treatment,and fluorescence labeling materials and their physical and chemical properties play a key role for the imaging quality.However,the fluorescent materials with aggregation induced emission(AIE)property are quite limited,and the fabrication and performance of fluorescence nanoparticles need to be further optimized.Moreover,the identification and tagging capabilities for specific tumor cells remain to be further improved.Aiming at the above issues,fluorescent nanoparticles(DPBA@PEG-PDLLA)with high quantum yield were prepared by using amphiphilic polymers(PEG-PDLLA)encapsulated with AIE dyes(DPBA).These bright nanoparticles with red emission could be applied to detect the cutting edge diagnosis in breast cancer through fluorescent bioimaging method.The research content of the study is as follow:(1)The analysis and imaging of the data of Raman spectroscopy obtained by mapping method in breast cancer and normal tissues.Collect the tissue of breast cancer and normal breast;freeze and fix the specimen.Two contiguous sections were excised using a freezing microtome.One section was stained with hematoxylin and eosin(H&E)for routine histopathological analysis;the other section was prepared for raman detection.We detect aim area(20?m×20?m)point by point by Lab RAM ARAMIS.Then we obtain 53 groups(2597 spectra)of mapping data;1280 spectra of point detection data.Firstly,mean Raman Spectra were get from mapping and point data.The characteristic peaks were analyzed.The peaks located at 1125,1307,1245,1445,1735cm-1 are belonging to lipid in normal breast tissues.But the peak only appear at 1124?1302?1446cm-1 in breast cancer tissues.The lipid is less in malignant tissue than normal.The peaks located at 967,879,1156,1582cm-1 are belonging to nucleic acid appeared in malignant tissue.But only 1157cm-1 appeared in normal breast tissue.The components of nucleus are higher in breast cancer tissue,and this is corresponding to the character of caner tissue-rich in nucleus materials and vigorous proliferation.By the analysis of the spectra of mapping and point data,we found outlier data is 12.7% in malignant mapping data,and 6.6% in normal tissues.It is 24.5% in malignant point data,and 26.0% in normal.There is obviously different between malignant mapping data and point data(t=-6.169,P<0.001)and also obviously between normal tissues' mapping data and point data(t=-8.873,P<0.001).It confirm that mapping data is more stable than point data,and these kinds of data have the advantage in the model building.Based on the mapping data,the diagnosis model build by KNN method provides excellent diagnostic power(accuracy 99.56%,sensitivity 96.6%,specificity 98.48%,PPV 99.56%,NPV 89.04%).At last we use two methods to image the detected area-model judgement and characteristic peaks.The first kind imaging is well corresponding to the pathology image.It may become a new method for the diagnosis of cutting edge in breast cancer operation.The second method use different intension of different characteristic peaks to image the area.The images at 1088?1215 and 1582 cm-1 are almost similar,and there is some degree of similar at 936 cm-1.This confirm that high expression of protein in cancer tissue is related to the cell division,metastasis and proliferation.This kind of image may become a kind of possible diagnosis method on molecular level.(2)Bright fluorescent nanoparticle for breast cancer MCF-7 imagingHerein,we develop a strategy for the fabrication of fluorescent nanoparticles with high quantum yield with bright red fluorescence,by using AIE active dye(DBPA)and amphiphilic polymers(PEG-PDLLA)severed as matrix.The nanoparticle DPBA@PEG-PDLLA show strong red emission peaked at 654 nm and high quantum yield up to 26% through optimizing the ratio of dye and polymer.Owing to the excellent properties of polymer,DPBA@PEG-PDLLA possesses uniform size with 88 nm,spherical morphology and good biocompatibility.The nanoparticles can stain in cytoplasm of MCF-7 cell with a strong red fluorescence signal,and pose little toxicity to living cell.More important,we fabricate multifunctional nanoparticles by using co-encapsulation approach,which employ DBPA as fluorescent probe and paclitaxel(PTX)as drug.The DPBA/PTX @PEG-PDLLA not only show bright red emission with quantum yield of 25%,but also possesses good drug release ability with cumulative release rate of 48 h up to 25.1%.The results of in vitro MTT and CLSM experiments showed that DPBA/PTX@PEG-PDLLA nanoparticles not only have good inhibiting ability with the cell proliferation of MCF-7 tumor cells,but also could be uptake by tumor cell and then absorbed by their cytoplasmic fluorescence imaging. |
PDF Full Text Request