Study Of Reverse Effect Of Xianhuayin In Lesion Precancerous Of Golden Hamster' Bucco-Mucosa Derivn By DMBA

Objective: The lesion precancerous of oral mucosa is a kind of precancerous state,showing off atypical hyperplasia of epithelium of oral mucosa in different extent,but it's not be the cancer.The new categorization of WHO give the definition of lesion precancerous as that:Precursor lesions are defined as altered epithelium with an increased likelihood for progression to squamous cell carcinoma(SCC).The altered epithelium shows a variety of cytological and archi-tectural changes that have traditionally been grouped under the term dysplasia. Currently accepted precancerous lesions and precancerous conditions, includingleukoplakia, erythema, lichen planus and oral submucous fibrosis, etc. Without timely treatment of precancerous lesions and left unchecked, will ultimately develop into cancer, seriously affect the treatment and prognosis, but also to patients a tremendous impact on physical and psychological. How to treat oral precancerous lesions, blocking its development to oral cancer, has become a hotspot of current research. At present for the oral treatment of precancerous lesions of large, mainly Western oral, systemic and local use of steroids and immunosuppressive agents, early surgical excision and traditional Chinese medicine treatment. But the efficacy of Western medicine instability,and hormones and immune inhibitors, although the early onset of fast but its role is not sustainable and side effects are large; the most previous tradition of Chinese medicine connect with application of local hormones,so there's little news about Chinese medicine treatment of precancerous lesions onlyand little study to prove the treatment effect of the Chinese medicine. Chinese medicine in the treatment of precancerous lesions of oral mucosa has obvious advantages for the characteristics of precancerous lesions of oral mucosa: Chinese medicine samples wen, effective solid; oral decoction or proprietary Chinese medicine preparation, taking the method is simple, patient compliance is good, easy to accept; with fewer side effects than western medicine and so on. In view of the above advantages, traditional Chinese medicine in the treatment of precancerous lesions of the increasing attention has been paid. In this study, drinking herbal Xianhuayin of experimental animals treated with oral precancerous lesion intervention, and by flow cytometry and immunohistochemistry to observe the before and after treatment P53 and COX-2 expression, analysis, and explore the Xianhuayin of drinking on the oral mucosa reversal of the role of precancerous lesions. For future clinical applications, experimental theory.Methods:1. Animal model of oral precancerous lesions and group preparationSyrian golden hamster get (SPF grade) 55, 6 to 8 weeks of age, body length of about 10cm, weight 90 ~ 110g, both male and female, from Beijing Tong Lihua Experimental Animal Technology Victoria Limited, the production license number: SCXK (Beijing) 2007-0001. In accordance with the principle of completely randomized into four groups: untreat control group, 5, precancerous lesions control group, 10, and saline-treat control group, 10, Xianhuayin-treat group, 30 to drink.The untreat control group, the normal daily feeding, untouched, until the end of the experiment, together with the other groups killed and drawn.Precancerous lesion control group, saline-treat control group, Xianhuayin-treat group are painted by 0.5% DMBA at the cheek pouch mucosa three times a week for six weeks.2. The brew of traditional Chinese medicine decoction of XianhuayinTraditional Chinese medicine in the first set of square by adding about 8 to 10 times as much water, soak 40 minutes to 1 hour, Wu Huo heat with 20 to 30 minutes to boil and maintain for about 10 minutes, and then about Huo Jian-Wen After 10 minutes, filtering, juice; then add the equivalent of six times the amount of the remaining residues in water, heated by Wu Huo 20 to 30 minutes to boil, with the text Huojian about 10 minutes, adding Amomum villosum, boil about 6 ~ 8 minutes, filtering, abandoned to the residues. Merging the two liquid, concentrated liquid until the required amount (including crude drug 1g/ml), packaging and sterilization, 4℃stored until required.3. Xianhuayin of traditional Chinese medicine treatment of drinking and saline gavageSaline-treat control group and Xianhuayin-treat group were treated with saline and drink decoction of Chinese medicine Xianhuayin gavage treatment, 1.6ml / only once a day for eight weeks , killed all the animals and subjects, fixed, microscopic examination.4. Immunohistochemical detection of P53 and COX-2 expression in all groupsTake wax blocks of each group, routine paraffin sections, dewaxing to water, in accordance with S-P kit instructions to operate, microscopic observation of the untreat control group, precancerous lesions control group,saline-treat control group and Xianhuayin-treat group, in the P53 and COX-2 expression, and analysis and comparison between the expression of each group whether there are differences, to evaluate the traditional Chinese medicine Xianhuayin precancerous lesions of oral mucosa reverse effect.5. Flow cytometryParaffin-embedded tissues on a continuous cut in the slicing machine to take the organization of 40-50μm thick film 5-8 tablets, placed in a test tube; xylene dewaxing, gradient alcohol hydration; saline wash, 120 mesh copper mesh Qingcuo organizations to collect a single cell suspension; 500 mesh nylon mesh filtration, centrifugation, 1000rpm, 5 minutes; to join an anti-working solution at room temperature, put it aside for 40 minutes; to join two anti-working fluid, put it aside for 1 hour at room temperature; machine inspection, read the results.6. Statistical MethodsUsing SPSS 13.0 statistical software, test results are the mean±standard deviation (±S) said that the application of analytical methods rank sum test P53 and COX-2 test results were statistically analyzed,α= 0.05, P <0.05 statistically significant.Results:1. Probabilistic selected specimens of 10 cases observed in the optical microscope, are found in irregular epithelial stratification, basal cell polarity loss, drop-shaped nail protrusion, pleomorphic nuclei, nuclear-cytoplasmic ratio increased, the nucleus increases, not like a typical split , nucleolus increases, increase in the number and nuclear deep-dyed so the performance of atypical hyperplasia, may indicate precancerous lesion models were successfully constructed.2. Immunohistochemistry2.1P53 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.2.2COX-2 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.3. FCM3.1P53 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.3.2COX-2 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.4. Correlation studies have shown that: P53 and COX-2 in oral precancerous lesions of the occurrence and development process, there is significant correlation.Conclusion:1.Applications lure cancer agent DMBA, in a certain time and frequency, it can be effectively induced hamster buccal mucosa precancerous lesions.2.P53 and COX-2 in oral precancerous lesions in the occurrence and development of great significance, P53 and COX-2 in oral precancerous lesions in the occurrence and development process, there is significant correlation.3.Medicine Xianhuayin in the treatment of precancerous lesions of drink has a noticeable effect.