| Gastric carcinogenesis is a complex and multistage biological process into which pre-cancerous lesions of gastric cancer (PLGC) were developted. To reverse the severity of epi-thelial dysplasia in gastric mucosa, it is very important to understand problems:early detec-tion, intervention and treatment of in patients with PLGC. PLGC mainly included intestinal metaplasia (IM) and dysplasia (Dys), both of which are risk factors in developing gastric can-cer, especially in the moderate or severe dysplasia.It is reported that both mild and moderate dysplasias were more likely to regress than to progress. The risk of progression from mild to severe dysplasia or worse was about15-30%, but the risk of progression from severe dyspla-sia was60-80%. How to effectively reverse the severity of gastric epithelial dysplasia is a good assay for prevention and treatment of gastric cancer.Under the concept of Chinese medicine and through the long term effort, our research group has put forward that based on deficiency of qi in spleen and stomach and/or deficiency of qi and yin, the pathogenesis of PLGC is attributed to symdrome of asthenia in origin and asthenia in superficiality and companied with the asthenia of blood stasis and toxin damage in stomach meridian and collaterals of stomach, whose basic treatment principals was Yiqi Huayu Jiedu. According to the principal of Yiqi Huayu Jied, Xiaopi granule is designed to supplement qi and nourish yin, promote blood circulation to remove blood stasis, and clear away heat and toxic material, which includes Codonopsis pilosula, Fried lily, Radix Linderae, Citron peel, Radix salviae miltiorrhizae, Rhizoma curcumae, Three seven powder, Dandelion, and Oldenlandia diffusa.Our clincal trials have proved that it has good therapeutic effect for not only significantly improving the clinical symptoms of patients with PLGC but also re-versing the the severity of gastric epithelial dysplasia.In this study, there are two sections:i.Literature research; ii.Experimental research re-spectively:i. With the method of Meta-analysis, we compared the effect, and pathological changes (the severity of PLGC and gastric epithelial dysplasia) between Chinese Herbal Medicine and Purely Western Medicine for PLGC, thus providing clinical evidence of higher quality for the treatment of traditional Chinese medicine and paid the foundation for next ex-perimental research; ii.Based on literature research and clinical trials on PLGC, it has further verified to have good impact on general conditions, macroscopic characterization and gastric epithelial pathological changes and in rats with chronic atrophic gastritis (CAG) with Dys under the treatment of Xiaopi granule, and make good advantage of modern molecular biolo-gy technology and method to investigate the epigenetic mechanism on how to reverse the se- verity of gastric epithelial dysplasia.a) Literature researchObjective:To compare the effect and pathological changes of patients with PLGC under the treatment between Chinese Herbal Medicine and Purely Western Medicine.Methods:After searching only randomized clinical trials comparing the purely Western Medicine between Chinese Herbal Medicine with using strategy (irrespective of blinding, searching with English and Chinese), two authors independently identified the trials for ex-clusion and inclusion criterion, extracted the data and assessed the methodological quality of the trials and risk of bias. Meta-analysis was performed by RevMan5.0to calculate the Risk Ratio (RR) and95%confidence intervals (CI) using Fixed-effects models or random-effects models.Results:4RCTs with250patients were included this review. There was statistically sig-nificant between two groups on overall effect[RR=1.52,95%CI(1.27,1.82)], moderate dys-plasia[RR=1.58, CI(1.09,2.29)]and precancerous lesions[RR=1.85,95%CI(1.35,2.54)]; but there was no statistically significant between two groups on severe dysplasia[RR=2.14,95%CI(0.84,5.41)]. Conclusion:The therapeutic effect of Chinese Herbal Medicine intervention of PLGC was thought to be significant over using the purely Western Medicine, but further randomized trials with high-quality and low-risked bias is needed.b) Experimental researchObjective:To establish the animal model of chronic atrophic gastritis with dysplasia in rats, to explore the intervention for general conditons, pathological changes in rats with CAG and Dys under the treatment of Xiaopi granule, further investigate the relationship between the methylation status of the5’CpG island of p16and Pten promoter region and the expres-sion of pi6and Pten protein of PLGC or GPL in rats.Methods:60SPF male Wistar rats were randomly divided into model group (50rats) and blank group (10rats). The blank group was given standard diet until the end of this experi-ment. The model group was intragastric administrated by120μg/mL MNNG (5mL/kg,1time/day), applied by0.05%aqueous ammonia for drinking water and feed by adding to0.03%ranitidine. The model was eatablished before the32th week ended. The remaining33rats of model group were randomly divied into four groups:i. Natural recovery group, ii.Vitamycin applided group, iii.Xiaopi grandule applided group. Each group has11rats, whcih was re-spectively intragastric administrated by3mL/kg saline,2mL/kg Vitamycin suspension liquid (0.3g/kg Vitamycin) and3mL/kg Xiaopi grandule suspension liquid (containing9g/kg raw material). All of those rats were killed after treatment for12weeks. The pathological chang-es(the severity of PLGC and gastric epithelial dysplasia), methylation status of CpG islands of tumor suppressor gene p16and Pten, and experssion of p16and Pten protein were observed with light microscope, Methylation Specfic PCR (MS-PCR), immunohistochemistry and oth-er means.Results:i. General situation (body weight, diet and water intake) and macroscopic characteriza-tion (fur color, Response ability, stool, tongue and et al) of rats in model group were worse than the blank group in modeling stage. In the intervention phase, general situation and macroscopic characterization of rats in Xiaopi Granule group and Vitamycin group were better than Natural recovery group, especially the Xiaopi granule group. There was no ob-vious recovery of general situation and macroscopic characterization in the natural recovery group.ii. It occured atrophic gastritis rats in model group on week26. On week28atrophy gastritis accompanied with intestinal metaplasia were observed in model group. On week30and32, it appeared with intestinal metaplasia, dysplasia, and no sign of recovery. Rats of Xiaopi granule group was better than that of rats of natural recovery model group in patho-logical changes (P<0.05), Rats of Vitamycin group was not better than the natural recovery group in the pathological changes of rats (P>0.05).iii. Correlation analysis between body weight and pathological changes of rats at en of this experiment, the Spearman correlation coefficient=-0.155, P>0.05, it negative that it has related changes of body weight with pathological changes.iv. The methylation positive status of CpG island of promoter region in p16tumor sup-pressor gene was detected among the blank group, natural recovery group, and Xiaopi granules group whose rates were16.7%,85.7%,50%,40%respectively. Through the posi-tive band calculating by image analysis software, it is shown that CpG island methylation level of of p16tumor suppressor gene in was significantly elevated than the natural recov-ery group (P<0.01).CpG island methylation level of p16tumor suppressor gene in Vitamycin group has statistical significance with natural recovery group (P<O.05). CpG is-land methylation levels of Xiaopi granule group in p16tumor suppressor gene had differ with Vitamycin group (P<O.05);v. The positive experssion of p16protein was observed among the blank group, natural recovery group, and Xiaopi granule group whose positive rates were100%,25%,55.6%and72.7%respectively. After intervention finished, the expression of P16protein in Xiaopi granule group was better than Vitamycin group (P<0.05) and natural recovery group (P<0.01), but with the blank control group, the expression of p16protein has remarkably statistical significance (P<O.01). vi.2cases of positive expression of p16protein were found in14cases of CpG island positive methylation of p16tumor suppressor gene; there was only1case of negative ex-pression of p16protein in16cases of CpG island negative methylation of p16tumor sup-pressor gene.Through two variables correlation test, Spearman coefficients was-0.800, P<0.01, it was thought that tumor suppressor gene p16CpG island hypermethylation was positively correlated with the expression of p16protein.vii. Binary logistic regression analysis showed p16tumor suppressor gene CpG island methylation status and expression of p16were statistically significant (P<0.01or P<0.05) with pathological changes, indicating they were the important factors influencing patho-logical changes.viii. CpG island positive methylation of promoter region in Pten tumor suppressor gene has not found, but the negative did. The positive expression of Pten protein was detect among blank group, natural recovery group, Vitamycin group and Xiaopi granule group whose positive expression rates were100%,50%,66.7%and81.8%respectively. After in-tervention finished, the positive expression of Pten protein Xiaopi granule group was better than Vitamycin group (P<0.01) and natural recovery group (P<0.01), and the positive ex-pression of Pten protein had no significant difference (P>0.05) with the blank group,ix. The two variable correlation analysis is shown that Spearman coefficient is-0.619, P<0.01, rats pathological changes was positively related to tumor suppressor gene Pten protein positive expression.Conclusion:i. Establishment of animal model with CAG and Dys in rats intragastric administrated by MNNG;ii. The Yiqi Huayu Jiedu principle for Xiaopi granule can effectively inprove the gen-eral situation, macroscopic characterization and gastric epithelial pathological changes and in rats with chronic atrophic gastritis (CAG) with Dys.iii. The Yiqi Huayu Jiedu principle for Xiaopi granule can reverse the severity of dys-plasia, decrease methylation level of CpG island of promoter region of suppressor tumor gene p16and upregulate positive expression of p16protein;iv. MSP has shown that there is no raltionshio between methylation status of CpG is-land of promoter region of suppressor tumor gene Pten tumor suppressor gene and gastric cancer development in rat;v. The Yiqi Huayu Jiedu principles for Xiaopi granule can upregulate positive expres-sion of Pten protein to reverse dysplasia.
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