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Background oral squamous cell carcinoma (oral squamous cell carcinoma, OSCC) is the most common malignant tumors of the oral and maxillofacial region, accounting for 3% to 5% of the systemic malignancies, in recent years, the incidence of oral cancer tend to be younger, young man oral fluctuations in the incidence of cancer in the range of 0.4% to 5.5%. Means of the development of oral cancer lesions to late after surgery, radiation therapy and chemotherapy, but both will have patients with local organizations structure damage, affect the the maxillofacial structure and function, brought great suffering to patients. Therefore, to study the occurrence of oral cancer development and prevention of oral cancer valued by the majority of scholars. Census of oral cancer high-risk groups is particularly important. Similar to human oral squamous cell carcinoma model of rat oral palatal mucosa, mucosal changes in the whole process of the bio-dye stained the naked eye and histological observation modeling, and lay the foundation for the early diagnosis of oral cancer. Syrian golden hamsters, rats, mice, animals used to establish an animal model of oral cancer. The reliability of the results of their research is limited due to the special anatomical and physiological environment of the cheek pouch great human oral mucosa difference. Scholars no longer meet in a simple model of the cancer establishment, but rather looking to establish the model of the human oral environment similar to more representative of oral cancer. Carcinogenesis model of oral cancer in the application of more animals are rats and mice. Scholars will be applied to the rear of 28 28-week-old Wistar rat hard palate palatal mucosa 4NQO, three times a week. A total of 19 weeks, induction the exit chamber palate mucosal cancer. Academics reported the hard palate in the rat oral mucosa smear-NQO1, 2, 4, 8, 12, 16 weeks to 24 weeks after feeding, visual observation of carcinogenesis eight weeks palatal rugae unclear boundaries a little change in the the animal hard palate mucosal carcinogenesis 12 weeks the animals showed obvious tumor. The histopathology results the animal hard palate mucosa carcinogenesis eight weeks for moderate or severe dysplasia, carcinogenesis 12 weeks the animals are well differentiated, squamous cell carcinoma horny. Ribeiro 0.005% 4NQO drinking water feeding Wistar rats induced well-differentiated squamous cell carcinoma in 20 weeks. Biological dyes, domestic and foreign scholars have been widely used in a variety of precancerous lesions and cancer laboratory examinations. Canto pointed out in the study reported on 43 patients with Barrett's esophagus lesions, methylene blue staining after biopsy than conventional random biopsy diagnosis specific columnar epithelial dysplasia and cancer with high accuracy. Dawsey study found that conventional endoscopy with 12ml / l Lugo staining endoscopy, Lugo staining endoscopy precancerous lesions and early invasive carcinoma sensitivity higher. In 1960, Sharwin first reported observe suspicious lesions of the mouth and throat, vital staining with toluidine blue (Toluidine Blue) identify benign and malignant. Oratest is a medicinal grade toluidine blue reagent, and its application has been reported, but the effect is uncertain. Present in animal oral mucosa carcinogenesis process dynamics of mucosal changes and biological dye staining, and further to find a relatively reliable biological dye reagent has not been reported. The purpose of this experiment is chemically induced rat oral mucosa of the human environment similar cancer animal model. Dye methylene blue combined with biological, Lugo the liquid carried noninvasive auxiliary diagnosis, the sensitivity of the mucous membrane of the comparison of both the process of carcinogenesis, and to choose a more suitable biological dye, auxiliary diagnostic tool for the early diagnosis of oral cancer. Method 1. The establishment of an animal model: all the animals were divided into two groups according to the random number table. The experimental group of 54 male Wistar rats, six per cage. Experimental group with a sterile cotton swab dipped in 0.5% 4NQO dimethyl sulfoxide solution 0.05ml ~ 0.1ml rubbed Wistar rats palate, three times a week, 24 weeks of continuous rubbed. The control group (n = 6) without applying any processing, the conventional drinking water diet. 10,13,16,19,21,24 weeks after the start of the experiment, according to the random number table is selected from the experimental group 8, select one from the control group were sacrificed. 2. In the modeling process, starting from 10 weeks to the experimental group and the control rats staining. Before the experiment, randomly selected according to the random number table eight experimental rats and a control group of rats were stained. Dyeing experiment was divided into two parts, the morning of the experimental group and the control rats methylene blue staining; afternoon experimental group and a control group of rats Lugo staining. Rinsed with distilled water before the experiment give a rat palate, dry, mucosal rubbed 0.5% methylene blue 1 ~ 3ml, 1 ~ 2min wipe with 1% acetic acid, and washed with distilled water three times in a row, dry, recording color disappears, observed mucosal changes the process of photography. Lugo liquid staining, washed with distilled water rat palatal mucosa, dry rubbed 1.5% to 3.0% Lugo liquid 1 ~ 3ml rubbed to the mucosa 1 to 2min after distilled water to observe the changes of the mucosa and the degree of staining and the scope of records the color disappears, photography. The animals were killed after the end of the experiment, cut specimens, immediately put fixed in 10% formalin fixative. Conventional sectioning, hematoxylin and eosin staining, Canada balsam Fengpian, light microscopy observations. 3. Judge: rat oral mucosa carcinogenesis criteria: rat oral mucosa pathological changes including epithelial dysplasia and oral cancer histological type diagnosed according to WHO (2005) standard. Judgment of the staining results: (1) evaluation of the results of methylene blue staining after 1% acetic repeated decolorization lesion and its surrounding tissue surface is still showing varying degrees of blue-positive staining (); records coloring degree (light blue, dark blue), homogenization (homogeneous coloring, blotchy, flaky coloring), and the stained area perimeter (perimeter clear perimeter unclear). Lesion and surrounding mucosa after bleaching blue light blue stranded or uniform negative staining (-). ② Lugol's iodine solution rubbed mucosal not homogeneous lightly stained or not stained positive; uniform brownish yellow coloring negative. 4. Application SPSS10.0 statistical software using x ~ 2 test, take α = 0.05 level for significance test. Results 1. The establishment of animal models: Casual extend the duration of action of carcinogenic agents, rat palatal mucosa white plaques, ulcers, erosions, granular hyperplasia changes have appeared. 10 weeks, 50.0% of rat palatal mucosa performance of simple epithelial hyperplasia, 50.0% for mild to moderate dysplasia; 13 weeks, 37.5% of mild dysplasia, 50.0% moderate dysplasia, severe abnormal proliferation of 12.5% : 16 weeks, 37.5% for mild to moderate dysplasia, severe dysplasia 37.5% and 25.0% for carcinoma in situ; 19 weeks with severe abnormalities and carcinoma in situ cases of invasive well-differentiated squamous cell carcinoma. 21 weeks, 37.5% moderate and severe dysplasia, 25.0% of carcinoma in situ, and 37.5% for invasive well-differentiated squamous cell carcinoma to cancer; 24 weeks, 87.5% of cancer, severe 12.5%. 2. Blue staining of oral cancer and epithelial dysplasia sensitivity of 88.9% and 80.8%, respectively; The Lugo staining of oral cancer and epithelial dysplasia sensitivity 55.6% and 50.0%, respectively. The positive staining rate of methylene blue and Lugo liquid with a significant difference (P <0.05) Conclusion 1. Application 4NQO carcinogenic agent partial erasure Wistar rat palatal mucosa oral the palatal mucosa cancerous animal model can be successfully established. 2. Rat palatal mucosa cancer is a dynamic process, experienced squamous epithelial hyperplasia, mild epithelial dysplasia, moderate epithelial dysplasia, severe epithelial dysplasia and carcinoma in situ, and further development to the early dip Stream cancer. 3. Methylene blue and Lugo staining, observed for the early diagnosis of oral epithelial dysplasia and cancer. 4. The lesion, methylene blue staining sensitivity better than Lugo liquid, methylene blue staining method for early diagnosis of oral cancer lesions.
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