| BackgroundEsophageal cancer is one of the most common malignant tumors, which is serious threat to human health and its overall five year survival rate is less than 20%. Therefore, the early diagnosis of esophageal cancer and reasonable treatment is very important to improve the quality of life in patients and prolong the survival time. In recent years, the applications of electronic endoscopy, chromoendoscopy, EUS, magnifying endoscopy, narrow band imaging, fluorescence endoscopy, confocal microscopy greatly improve the diagnosis rate of esophageal cancer. But the final diagnosis still pathological diagnosis. Preoperative biopsy may cause hemorrhage, needle, metastasis, clinical pathological process time-consuming disadvantages and other shortcomings; while the postoperative histopathological diagnosis, need material and subsequent tissue fixation, embedding, sectioning, staining. Multiphoton microscopy (MPM) technology of nonlinear optics and laser on femtosecond laser multiphoton "optical biopsy" based on the second harmonic, produced by cells using tissue autofluorescence and itself produces collagen, can quickly obtain real-time tumor tissue structure, cell morphology and tumor stroma in tumor invasion changes in collagen transfer reaction. Autofluorescence cells generated from intracellular nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD). Characteristics of multiphoton "optical biopsy" can not only capture the tumor tissue, but can reflect the metabolic level of tumor cells and the surrounding stroma collagen changes in signal. MPM without the traditional pathology of multiple steps, is a real-time and fast diagnosis method, with real-time, nondestructive, high penetration rate and high resolution. MPM is expected to become a real-time diagnostic tool for cancer.Objective:1. Distinguish among normal esophageal tissues, cancer tissues, as well as cancer tissues after paclitaxel elutin stent implantation under MPM;2. Summarize the imaging characteristics of human normal esophageal tissue, esophageal intraepithelial neoplasia and esophageal squamous cell carcinoma under MPM;3. Using the above summed up the characteristics to compute the sensitivity of, specificity and accuracy MPM diagnostic efficacy.Methods:1.A total of 24 New Zealand white rabbits were purchased from Nanjing Qinglongshan animal breeding farms, Certificate of Conformity:SCXK (Su) 2007-0008. The heathy rabbits were 3 months age, weighting 1.8-2.0kg with grain feed, VX2 (rabbit squamous epithelial cancer) tissue blocks injected into rabbit hind leg muscle, about two weeks later grow into cancer tumors, the tumor removed under sterile conditions and was cut into small pieces frozen in-80 ℃ ultra-low temperature freezer;2. The tumor tissue was injected into rabbit esophagus:endoscopic animal model was made by the tissue blocks taken out of recovery, cut to a size of about 1mm, through injection needle in endoscopic tissue injected into the submucosa of esophagus. The superfine endoscopy determined more than 2/3 to meet the narrow lumen of stent implantation indications, the stent stent implantation by conveyer lesion. The rabbits were dissected after two weeks;3. Divide the normal rabbit esophagus, esophageal tissue lesions of esophageal cancer tissue and tumor after stenting into three groups. The tissue was frozen section. Collect the MPM image and H-E image.Compare the imaging features in the three groups;4. Collect the human normal esophageal tissue, high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma, according to the above method for MPM imaging and H-E image. Summed up the characteristics of MPM imaging in the three groups;5. Expand the sample size, using the above summed up the characteristics to compute the sensitivity, specificity and accuracy of MPM diagnostic efficacy.Results:1. Summed up the MPM imaging features of the normal rabbit esophagus, esophageal cancer and esophageal cancer of stent after treatment successfully, The mean optical density of collagen for normal esophageal tissue was16.15 ± 5.73, tumor group was 1.46±1.20, stenting group was4.16 ± 2.17, P<0.05, the difference was statistically significant;2. MPM could distinguish between normal esophageal tissue, high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma. The ratio of NADH/FAD in esophageal squamous cell carcinoma was 3.36±0.28, in esophageal high-grade intraepithelial neoplasia was 2.85 ± 0.19 and in normal esophageal tissues was 1.91 ± 0.12, P<0.05, the difference was statistically significant;3. The MPM diagnostic efficacy:sensitivity:95%, specificity:94.44%, the positive predictive value of 97.08%, negative predictive value of 90.67%, accuracy:94.81%.Conclusions:1. MPM can distinguish the normal esophageal tissue, esophageal cancer tissues and tissues after stent implantation;2. MPM can diagnose normal esophageal tissue, esophageal intraepithelial neoplasia and esophageal carcinoma tissue;3. The diagnostic value of MPM is relatively high, can achieve real-time diagnosis, has considerable development foreground. |
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