【作者】 王彦刚；

【导师】 李佃贵；

【作者基本信息】 河北医科大学 ， 中西医结合临床， 2009， 博士

【摘要】 慢性萎缩性胃炎（chronic atrophic gastritis,CAG）是消化系统的一种常见病,是指炎症导致的胃腺体破坏、减少、萎缩、甚至消失,表面上皮细胞丧失分泌能力,黏膜肌层增厚的病理、临床病症。上世纪40年代Schindle首次提出CAG的概念,1978年WHO将其列为胃癌的癌前疾病或癌前状态（Precancerous conditions）,而在CAG基础上伴发的肠上皮化生（Intestinal metaplasia,IM）和异型增生（Dysplasia,Dys）,则是胃癌的癌前病变（Precancerous Lesions of Gastric Cancer,PLGC）,癌变的发生率较高。CAG及其癌前病变已成为目前研究的重点,及早识别、防治癌前疾病和癌前病变,成为降低胃癌发生率和死亡率行之有效的方法。目前,西医学治疗本病尚缺乏有效的治疗手段。中医药在治疗CAG方面有着明显的优势,其改善患者症状及预防疾病向癌转化等方面的疗效肯定且显著。中医学将CAG病变归属于“胃脘痛”、“胃痞”、“痞满”、“嘈杂”等病证范畴。导师李佃贵教授认为,随着社会的发展,生活方式、饮食结构的改变,环境因素变化,包括大气污染等,现代人的精神社会压力加大,疾病模式会发生变化。在临床诊疗中观察到许多慢性疾病患者病情反复,缠绵难愈,常有颜面晦浊、暗滞,少泽,大便粘腻不爽,小便黄,舌质暗红,苔黄或黄腻,脉弦滑等脉证表现,李师在总结前人理论的基础上,结合多年的临证经验,提出“浊毒学说”,认为浊毒既是一种对人体脏腑经络及气血阴阳均能造成严重损害的致病因素,同时也是多种病因导致脏腑功能紊乱、气血运行失常,机体内产生的代谢产物不能及时正常排出,蕴积体内而化生的病理产物。其致病特点,（1）粘滞难解易阻遏气机;（2）入血入络易伤气阴;（3）气血失调易瘀易积。浊毒为患所造成的病证,属于浊毒证,是指以浊毒为病因使机体处于浊毒状态从而产生特有临床表现的一组或几组证候群。运用浊毒学说指导CAG治疗,初步取得了较为满意的临床疗效。本研究旨在进一步完善该学说在CAG的证候规律,探讨化浊解毒方药的作用机制。目前,中医关于CAG的证候分型,行业学会及各家经验存在多种不同的观点,如有肝胃不和、脾胃湿热、湿浊中阻、胃络瘀阻、脾胃虚弱、胃阴不足6个证型分类;有肝胃不和、脾胃虚弱（含虚寒）、脾胃湿热、胃阴不足、胃络瘀阻、脾虚气滞等证型分类;还有的分为气阴两虚,虚火灼胃和脾胃气虚证型等等,繁杂不一,为临床和科研工作带来极大不便。中医证候学研究是目前中医药研究工作的前沿领域,20年来,随着统计学、流行病学、计量学、模糊数学等诸多学科知识的不断渗透,被用于中医证候诊断标准量化研究的方法也日益增多。聚类分析是将样品个体或变量指标按其具有的特性进行分类的一种多元统计方法,分为样品聚类分析和变量聚类分析,其目的是根据样品或指标间相似程度的大小,将性质相近的归在同一类,而将性质相差比较大的归在不同的类,在中医证候研究中的合理应用能够极大地提高中医辨证的规范性和准确性。本研究采用聚类分析的方法,分析CAG较大样本病例的临床表现,以期为临床提供较为合理、科学、规范的辨证分型,同时,通过应用本方法来探讨浊毒证的证候规律。观察CAG浊毒证患者的胃泌素（Gas）、胃动素（MTL）、生长抑素（SS）水平变化,探讨CAG浊毒证证型与胃肠激素之间的相关关系,进一步明确胃肠激素的变化在CAG中医证型中是否有特异性,胃肠激素变化异常是否能导致不同证型临床征象,提高对本证型的认识。CAG属癌前疾病,CAG伴IM,被认为是萎缩的典型标志以及胃癌的前兆,通过上述分析,发现浊毒内蕴是本病的主要证候特点,为此,创立了化浊解毒方,并观察本方治疗CAG的临床疗效。通过观察化浊解毒方药及其拆方含药血清对体外培养的人胃癌BGC-823细胞增殖和凋亡的影响,从实验研究方面,进一步探讨化浊解毒方药在防治胃癌前病变中可能的作用机制。第一部分慢性萎缩性胃炎证候学聚类研究目的:采用变量聚类分析的方法,对CAG临床资料进行探索性的中医证候分类研究,旨在应用现代数理统计方法探索CAG较为规范的临床证候分型,验证浊毒证候的特点,为临床辨证提供客观依据。方法:采集河北省中医院消化科门诊及住院CAG患者,所有患者依照西医诊断标准确诊为CAG,符合纳入、排除标准。记录371例CAG患者的症状、体征、舌象和脉象等临床资料,使用SAS软件对56个变量进行3-6类的聚类分析。结果:确定CAG患者中医证候分为:浊毒内蕴、肝胃不和、胃络瘀阻、脾胃虚弱、胃阴不足5类,其解释比例为72.74%,并初步确定了各证候诊断的主症、次症。浊毒内蕴证诊断:主症:畏寒肢冷,胃脘隐痛,苔黄、厚、腻,脉细、滑;次症:胸闷,胁胀痛,口臭,脉沉、缓。肝胃不和证诊断:主症:胃脘胀满,嗳气,脉弦;次症:嘈杂,反酸,易怒,饮食减少,嗳腐吞酸。胃络瘀阻证诊断:主症:胃脘刺痛,拒按,面色暗滞,脉涩;次症:恶心,舌有瘀点、瘀斑,苔薄。脾胃虚弱证诊断:主症:胃脘痞满,气短,便溏,舌有齿痕;次症:面色萎黄,腹痛欲泻,乏力,舌淡,脉弱。胃阴不足证诊断:主症:胃脘胀痛,苔少或无,舌少津,舌有裂纹,大便干,脉数;次症:口干,失眠,脉迟。小结:应用变量聚类分析,同时结合临床专家意见,能够帮助进行CAG中医证候的合理分类,并对提出的CAG新的证候类型—浊毒内蕴证进行了较为规范、科学的验证,为中医辨证论治CAG提供一定较为合理、科学的理论依据。第二部分慢性萎缩性胃炎浊毒证与胃肠激素相关性研究目的:本研究旨在通过探讨CAG浊毒证与胃肠激素分泌变化规律相关性研究,以揭示CAG浊毒证患者胃肠激素的分泌特点,提高对浊毒证的认识,为临床诊治提供客观指标方面的依据。方法:采集河北省中医院2006年12月-2007年12月消化科门诊及住院的CAG患者50例,所有患者据符合西医诊断标准和中医辨证诊断标准,分为浊毒证组25例和脾虚证组25例,同时选取健康志愿者设立健康对照组25例。采用放射免疫法检测患者血浆胃泌素（Gas）、胃动素（MTL）、生长抑素（SS）的含量。结果:1血浆Gas水平与健康组比较,CAG浊毒证组患者血浆Gas水平无明显变化,两组之间比较,差异无统计学意义（104.82±47.91 pg/ml VS 100.19±48.76 pg/ml, P>0.05）。与健康组比较,CAG脾虚证组患者血浆Gas水平均下降明显,两组之间比较,差异有统计学意义（79.17±34.02 pg/ml VS 100.19±48.76 pg/ml, P<0.05）。与CAG脾虚证组比较,CAG浊毒证组患者血浆Gas水平较高,两组之间比较,差异有统计学意义（104.82±47.91pg/ml VS 79.17±34.02 pg/ml, P<0.05）。2血浆MTL水平与健康组比较,CAG浊毒证组患者血浆MTL水平无明显变化,两组之间比较,差异无统计学意义（302.89±126.82 pg/ml VS 326.00±58.76 pg/ml, P>0.05）。与健康组比较,CAG脾虚证组患者血浆MTL水平下降明显,两组之间比较,差异有统计学意义（208.72±85.21 pg/ml VS 326.00±58.76 pg/ml, P<0.05）。与CAG脾虚证组比较,CAG浊毒证组患者血浆MTL水平较高,两组之间比较,差异有统计学意义（302.89±126.82pg/ml VS 208.72±85.21 pg/ml, P<0.05）。3血浆SS水平与健康组比较,CAG浊毒证组患者血浆SS水平无明显变化,两组之间比较,差异无统计学意义（11.81±6.34 pg/ml VS 10.10±7.20 pg/ml, P>0.05）。与健康组比较,CAG脾虚证组患者血浆SS水平下降不明显,两组之间差异无统计学意义（10.95±6.87 pg/ml VS 10.10±7.20 pg/ml, P>0.05）。与CAG脾虚证组比较,两组血浆SS水平之间,差异无统计学意义（11.81±6.34pg/ml VS 10.95±6.87 pg/ml, P>0.05）。小结:1在CAG浊毒证患者中,血浆Gas、MTL的变化有证型特异性。2血浆Gas、MTL的变化可能导致不同证型临床征象。第三部分化浊解毒方治疗慢性萎缩性胃炎临床研究目的:本文旨在通过观察化浊解毒方对CAG患者的症状积分、胃镜像、病理学、Hp等方面的影响,探讨该方治疗CAG的临床疗效。方法:收集河北省中医院消化科门诊及住院的CAG患者共92例,随机分为治疗组和对照组,治疗组口服化浊解毒方治疗,1袋/次,2次/日,早、晚空腹温服。对照组口服中成药胃复春4片/次,3次/日。12周为1个疗程,连用2个疗程。结果:1两组临床疗效比较:治疗组52例中临床痊愈17例,显效21例,有效11例,无效3例,总有效率94.23%;对照组40例中临床痊愈7例,显效10例,有效8例,无效15例,总有效率62.5%。两组临床疗效比较,差异有统计学意义（P<0.01）。2两组主要症状积分比较:两组治疗后胃痛、痞满、嗳气、纳差、乏力等症状积分明显减少,差异有统计学意义（治疗组:1.32±0.51 VS 4.82±1.25 P <0.01;对照组:1.58±0.65 VS 4.69±1.17, P <0.01）;治疗后,治疗组症状积分明显低于对照组,与对照组比较,差异有统计学意义（1.32±0.51 VS 1.58±0.65, P<0.05）。3两组胃镜疗效比较:治疗组胃镜结果显示在胃黏膜充血、糜烂、黏膜白象、黏膜粗糙、血管透见方面的疗效优于对照组,两组比较,差异有统计学意义（治疗组总有效率分别为86.96%、92%、64.58%、71.42%、65.63%,对照组总有效率分别为60%、78.95%、38.23%、36.36%、31.82%,P<0.05或P<0.01）。治疗组和对照组改善胃黏膜水肿的比较,差异无统计学意义（治疗组总有效率为87.5%、对照组总有效率71.88%,P>0.05）。4两组病理疗效比较:治疗组在改善胃黏膜萎缩,肠上皮化生（IM）总有效率分别为78.85%、84.62%,对照组总有效率分别为52.5%、55%,治疗组明显优于对照组,两组比较,差异有统计学意义（P<0.01）。5两组抗Hp感染疗效比较:治疗组和对照组抗Hp感染的根除率分别为47.22%和44.44%,两组的根除率比较,差异无统计学意义（P>0.05）。6安全性指标检测:患者治疗前后血、尿、便常规及肝、肾功能和心电图的均无异常改变。小结:1化浊解毒方能够较好改善CAG患者的临床症状。2化浊解毒方对胃黏膜的恢复、肠上皮化生的改善有较好疗效。3化浊解毒方有一定的抗Hp感染作用。第四部分化浊解毒方药及其拆方含药血清对人胃癌BGC-823细胞增殖和凋亡的影响目的:本课题旨在通过观察化浊解毒方药及其拆方含药血清对人胃癌BGC-823细胞增殖和凋亡的影响,探讨该方药是否通过抑制细胞增殖作用机制,预防CAG癌前病变,为该方药临床应用提供实验依据。方法:1含药血清的制备:SD大鼠,雄性,随机分为4组:空白对照组、化浊解毒方组、化浊方组、解毒方组。实验组分别按345.9g·kg^-1·d^-1、208.4g·kg^-1·d^-1、137.3g·kg^-1·d^-1剂量灌胃,空白对照组2ml生理盐水灌胃,1次/日,共3次。末次给药后2h,常规分离血清保存。2给药及分组:分别以各组的含药血清对胃腺癌BGC-823细胞进行干预,分为空白对照组、化浊解毒方组、化浊方组、解毒方组4组。3检测指标:分别采用MTT、流式细胞术、免疫组化法检测不同含药血清对BGC-823细胞的生长抑制、细胞周期和凋亡率的影响及BGC-823细胞中Bcl-2、Bax蛋白的表达。结果:1细胞增殖抑制实验:与空白对照组比较,化浊解毒方、化浊方、解毒方,均对BGC-823细胞有抑制作用（P<0.05或P<0.01）,抑制率分别为39.3%、16.5%、16.8%。尤以化浊解毒方效果显著,其抑制作用强于化浊方和解毒方（P<0.01）,而化浊方组与解毒方组之间比较,差异无统计学意义（P>0.05）。2 FCM对BGC-823细胞的细胞周期检测结果:与空白对照组比较,经化浊解毒方处理的BGC-823细胞的细胞周期发生明显变化,G0/G1期细胞明显增多（P<0.01）,S期细胞比率显著下降（P<0.01）,G2/M期无明显变化（P>0.05）,多数细胞阻滞在G0/G1期。而化浊方、解毒方对BGC-823细胞的细胞周期无明显影响（P>0.05）。3 FCM对BGC-823细胞凋亡率的检测结果BGC-823细胞经不同含药血清处理48h后,DNA直方图上出现典型的亚二倍体凋亡峰。FCM结果显示:化浊解毒方组、化浊方组、解毒方组凋亡率分别为（16.85±0.63）%、（11.48±0.50）%,（12.38±0.60）%,与空白对照组（2.09±0.25）%比较,差异均有统计学意义（P<0.01）;与化浊解毒方比较,化浊方组、解毒方组凋亡率有统计学意义（P<0.01）;化浊方组与解毒方组比较,差异无统计学意义（P>0.05）。4免疫组化检测Bcl-2、Bax蛋白表达BGC-823细胞经化浊解毒方、化浊方、解毒方含药血清干预48h后,Bcl-2蛋白阳性表达率均明显降低,分别为（29.73±4.34）%、（41.63±6.47）%、（37.11±4.63）%,与空白对照组（53.74±2.97）%比较,差异均具有统计学意义（P<0.01）。化浊方组、解毒方组与化浊解毒方组比较,差异有统计学意义（P<0.01）,而化浊方与解毒方两组比较,差异无统计学意义（P>0.05）。Bax蛋白阳性表达率均显著升高,化浊解毒方组、化浊方组、解毒方组阳性表达率分别为（58.11±4.19）%、（32.25±2.93）%、（34.84±2.66）%,与空白对照组（25.17±2.36）%比较,差异均有统计学意义（P<0.01）。化浊方组、解毒方组与化浊解毒方比较,差异有统计学意义（P<0.01）,而解毒方与化浊方两组比较,差异无统计学意义（P>0.05）。5流式细胞术检测Bcl-2、Bax蛋白表达流式细胞仪检测结果显示,与空白对照组比较,化浊解毒方、化浊方、解毒方作用于胃癌BGC-823细胞48h后,Bcl-2蛋白表达均明显降低,具有统计学意义（257.67±9.02 VS 330.22±7.13;291.22±11.07 VS 330.22±7.13;276.20±9.10 VS 330.22±7.13 ,P均<0.01）;细胞内Bax蛋白表达水平均增高,具有统计学意义（330.14±8.19 VS 281.23±12.71,P<0.01;303.84±6.60 VS 281.23±12.71, P<0.05;310.54±13.29 VS 281.23±12.71,P<0.01）。Bcl-2蛋白和Bax蛋白表达水平,与化浊解毒方组比较,解毒方组、化浊方组差异均有统计学意义（分别为P<0.01,P<0.05）;解毒方组与化浊方组比较,差异无统计学意义（P>0.05）。小结:1化浊解毒方可抑制人胃腺癌BGC-823细胞增殖,其作用机制可能与其干扰细胞周期,诱导细胞凋亡有关。2化浊解毒方及其拆方发挥诱导凋亡作用的机制可能是通过上调促凋亡蛋白Bax的表达,下调凋亡抑制蛋白Bcl-2的表达来实现的。3化浊方、解毒方均能诱导细胞凋亡,而对细胞周期无影响。经两者配伍后的化浊解毒方不仅能诱导细胞凋亡,而且能将细胞阻滞于G0/G1期,因此其对胃腺癌BGC-823细胞抑制增殖作用明显增强。结论:1应用变量聚类分析,能够帮助进行CAG中医证候的合理分类,并探讨CAG浊毒证证候特点,为中医辨证论治CAG提供一定较为合理、科学的理论依据。2在CAG浊毒证患者中,血清Gas、MTL的变化有证型特异性。血浆Gas、MTL的变化可能导致不同证型临床征象。3化浊解毒方药能够较好的改善CAG患者的临床症状,对胃黏膜的恢复、肠上皮化生的改善亦有较好疗效,并且有一定的抗Hp感染作用。4化浊解毒方及其拆方含药血清对人胃癌BGC-823细胞增殖产生抑制作用,促进BGC-823细胞的凋亡,可能是该类方药预防胃癌及逆转癌前病变作用机制之一。更多还原

【Abstract】 Chronic atrophic gastritis（CAG） is a common disease of digestive system. Its pathological changes was caused by inflammation, such as the destruction, reduce, shrink or even disappear of gastric glands, the loss of surface epithelial cell secretion capacity and mucosal thickening of the muscle. The last century 40’s, Schindle first proposed the concept of CAG. WHO would be the CAG as a precancerous lesion of gastric cancer in 1978, CAG associated with intestinal metaplasia and dysplasia are precancerous lesions of gastric cancer. The possibility of cancer is larger. CAG and its precancerous lesions has become the focus of the study. Early detection, prevention and treatment of precancerous diseases and precancerous lesions, become a effective way of lower incidence and mortality of gastric cancer.The application of traditional Chinese medicine（TCM） in the treatment of chronic atrophic gastritis has a distinct advantage, the improvement of patient symptoms and to correct the effect of disease and so on, it is affirmative and significant.CAG disease would be attributed to "Epigastralgia, gaster-mass, consumptive disease" syndrome in TCM. Professor Diangui Li thinks that modern people’s pressure of spirit and society become larger and disease patterns will changes with the development of society, lifestyle and diet changes. We observed many patients with chronic diseases that their conditions were repeated and refractory lingering in the clinical treatment.They had several clinical manifestations,such as dark cloud complexion, dark stagnation and less burnish, sticking and greasy stool, yellow urine, dark red body of the tongue, surname or greasy coated tongue, slippery pulse and so on. Professor Diangui Li introduced Zhuodu theory in the base of summarizing predecessor’s theory combined with years of clinical experience .He think Zhuodu is one of pathopoiesis factors which can cause serious harm on the human entrails and meridian and the QI-blood of yin and yang. Meanwhile, it’s the pathological products came from metabolic products which accumulated in the body when it’s entrails functional disorders and QI-blood circulation is not normal.Three pathogenic characteristics of Zhuodu: （1）Zhuodu is hard to dissolve and easy inducing disorder of vital energy.（2） Zhuodu is easy entering blood and collaterals and inducing disorder of vital energy.（3） Zhuodu is easy inducing incoordination between vital energy and blood and forming blood stasis. The symptoms caused by Zhuodu belong to Zhuodu Zheng. It refers to a group or several groups of syndromes group which have specific clinical manifestations when the body is in the state of Zhuodu caused by Zhuodu.We have obtained a relatively satisfactory clinical efficacy applying Zhuodu theory in the treatment of CAG. The study aims at consummating further the symptoms law of the theory in CAG and exploring the mechanism of Huazhuojiedu prescription.At present, it is discordant that the viewpoint of many institute and medical scientist about the typing of CAG in TCM. Some Chinese medical scientist consider that the typing of CAG is GanWeiBuHe, PiWeiShiRe, ShiZhuoZhongZu, WeiLuoYuZu, PiWei XuRuo, WeiYingBuZu. But other Chinese medical scientist considered that the typing of CAG is GanWeiBuHe, PiWeiXuRuo, PiWeiShiRe, WeiYingBuZu, WeiLuoYuZu, PiXuQiZhi. At the same time, some Chinese medical scientist consider that the typing of CAG is QiYing LiangXu, XuHuoZuo Wei, PiWeiQiXu. The discrepancy about the typing of CAG in TCM increase difficulty in evolving clinical work and scientific research.The study about the symptom of TCM is the frontline of the research on TCM. Over 20 years, the investigative methods of quantizating applicated in studying on diagnostic criteria of Chinese medical symptom are increasing along with the permeating of the knowledge of statistics, epidemiology, metrology, vague mathematics and so on. Cluster analysis is one kind of multivariant statistical method whice can sort the sample or variables index to sample cluster analysis or variables cluster analysis according to theirs characteristics. Cluster analysis aims to classify those samples or variables index whice have similar character into one sort according to the similar degree among the samples or variables indexs. Reasonable application of cluster analysis in study on Chinese medical symptom can enormously raise specification and accuracy rating of differentiation of symptoms and signs in TCM.In this study, we have adopted the way of cluster analysis to analysis of large sample of cases of clinical manifestations and aim to provide reasonable scientific and normative differentiation of symptoms and signs for classification of syndrome for clinical, to investigate the regularity of symptoms and signs of Zhuodu Zheng.We tried to explore correlativity of the Zhuodu Zheng with the gut hormones by observing the change of the level of Gas, MTL and SS in patients of CAG and definite if the change of the level of gut hormones have variability in different type of syndrome of TCM, and if the abnormal content of gut hormones can lead different type of syndrome. The study can elevate our recognition about the Zhuodu Zheng of CAG.CAG is a kind of premalignant disease. CAG combining IM was known as atrophic typical indicate and gastric carcinomatous premonition. Through above-mentioned analysis, it was discovered that Zhuodu NeiYun was the principal characteristic of symptom of CAG, so we formulated HuaZhuoJieDu decoction and observed the curative effect of Huazhuo Jiedu decoction on treating CAG combining IM.At experimental point of view, we explored the possible mechanism of Huazhuo Jiedu to prevent and cure gastric carcinomatous precancerosis by observing the effect of Huazhuo Jiedu and their separate Huazhuo Jiedu decoction containing serum on proliferation and apoptosis of cultured human gastric cancer BGC-823 cell in vitro.Part 1: Study on Symptoms of Chronic atrophic gastritis in Traditional Chinese medicine by the way of Cluster analysis Objective:The study on the clinical data of CAG probed the regularity of different type of syndrome of TCM of CAG by the way of Cluster analysis. The study aims at exploring normative typing of syndrome of TCM of CAG and providing objective evidence for clinical differentiation of symptoms and signs by the way of modern mathematical statistics.Methods: The 371 patients visiting the clinic service or being in hospital at the Medical Department of Digestion of TCM Hospital of Hebei Province were diagnosed CAG according to the diagnostic criteria in modern medicine. Those patients are accordance with the criteria of internalizing and removing. The sysptoms,physical sign, tongues demonstration and pulse tracings of the 371 patients of with CAG were collected and recorded. We did Cluster analysis by SAS software for 56 variables through sorting 3-6 types.Results: The syndromes of these CAG Patients could be divided into 5 types: ZhuoDuNeiYun, GanWeiBuHe, WeiLuoYuZu, PiWeiXuRuo, WeiYing BuZu. The proportion explained was 72.74%. The principal and minor symptom of each type of syndromes of these CAG initially definited. The diagnostic criteria of ZhuoDuNeiYunZheng: the principal symptom and sign: chilly and cold limbs, stomachache, yellow thick and greasy fur,samll and smooth pulse; the minor symptom and sign: full sensation in the chest, distending pain over hypochondrium, halitosis, sunken pulse, even and soft pulse. The diagnostic criteria of GanWeiBuHeZheng: the principal symptom and sign: distention of stomach, eructation, wiry pulses; the minor symptom and sign: gastric upset, sour regurgitation, ill-temper, anorexia, eructation with foul odour and acidr regurgitation. The diagnostic criteria of WeiLuoYuZuZheng: the principal symptom and sign: prickle of stomach and tenderness, dimmish blackish complexion ,unsmooth pulses; the minor symptom and sign: nausea, tongue with ecchymosis, thin fur;The diagnostic criteria of PiWeiXu RuoZheng: the principal symptom and sign:felling of fulless and oppression, shortness of breath, loose stool, tongue with teeth marks on its margin; the minor symptom and sign:s,allow complexion, abdominal pain, acratia, pale tongue, weak pulse. The diagnostic criteria of WeiYing BuZuZheng: the principal symptom and sign: distending pain of stomach, tongue without fur, fissuared tongue, constipation, ripid pulses; the minor symptom and sign: dry mouth, sleepessness, slow pulse.Conclusions: The way of variable cluster analysis combined with the opinions of clinical expert could help us do reasonable syndrome differentiation for CAG patients. Meanwhile, ZhuoDuNeiYunZheng was inspected and verified rationally and scientifically as one new type of syndrome of CAG by the study. The study provides certain reasonable and scientific theoretical foundation for CAG on selection of treatment based on the differential diagnosis.Part 2: Study on dependablity of Zhuo Du Zheng of Chronic Atrophic Gastritis with Gut HormoneObjective: The study aims to reveal the characteristic of gut hormone secretion of the patient of CAG Zhuo Du Zheng and to provid objective evidence on indicatrix by investigating the dependablity of Zhuo du zheng with the changing of gut hormone.Methods: 50 patients who visited the Medical Department of Digestion of Traditional Chinese Medicine Hospital of Hebei Province between December, 2006 and December, 2007 were diagnosed CAG according to diagnostic criteria in modern medicine. Those patients who lived up to the criteria to internalize were divided into Zhuo Du group and Pi Xu group according to diagnostic criteria in Traditional Chinese Medicine. Healthy normal group also were composed of 25 healthy volunteers. The number of people in three groups all was 25.The level of gastrin, motilin and somatostatin in blood plasma were detected by radio-immunifaction.Results:1 The level of Gas in blood plasmaCompared with normal group, the level of gastrin in blood plasma of Zhuodu group didn’t have marked change, there wasn’t statistical significance between two groups（104.82±47.91 pg/ml VS 100.19±48.76 pg/ml, P>0.05）.Compared with normal group, the level of gastrin in blood plasma of Pixu group remarkably decreased. There was statistical significance between two groups（79.17±34.02 pg/ml VS 100.19±48.76 pg/ml, P<0.05）.Compared with Pi Xu group, the level of gastrin in blood plasma of Zhuodu group was higher. There was statistical significance between two groups（104.82±47.91pg/ml VS 79.17±34.02 pg/ml, P<0.05）.2 The level of MTL in blood plasmaCompared with normal group, the level of motilin in blood plasma of Zhuodu group didn’t have marked change, there wasn’t statistical significance between two groups（302.89±126.82 pg/ml VS 326.00±58.76 pg/ml, P>0.05）.Compared with normal group, the level of motilin in blood plasma of Pixu group remarkably decreased. There was statistical significance between two groups（208.72±85.21 pg/ml VS 326.00±58.76 pg/ml, P<0.05）.Compared with Pixu group, the level of motilin in blood plasma of Zhuodu group was higher. There was statistical significance between two groups（302.89±126.82pg/ml VS 208.72±85.21 pg/ml, P<0.05）.3 The level of SS in blood plasmaCompared with normal group, the level of somatostatin in blood plasma of Zhuodu group didn’t have marked change, there wasn’t statistical significance between two groups（11.81±6.34 pg/ml VS 10.10±7.20 pg/ml, P>0.05）.Compared with normal group, the level of somatostatin in blood plasma of Pixu group didn’t decreased. There wasn’t statistical significance between two groups（10.95±6.87 pg/ml VS 10.10±7.20 pg/ml, P>0.05）.Compared with Pixu group, There wasn’t statistical significance between Zhuodu group and Pixu group（302.89±126.82pg/ml VS 208.72±85.21 pg/ml, P>0.05）.Conclusions:1 The level of gastrin and motilin in blood plasma have variability in different type of syndrome of TCM of CAG .2 The change of the level of gastrin and motilin in blood plasma may be the reason to induce the different clinical manifestation among different type of syndrome.Part 3: Observated the clinical curative effect of chronic atrophic gastritis with intestinal metaplasia based on Zhuodu theoryObjective:This research aimed to observe clinical effect of treating chronic atrophic gastritis with intestinal metaplasia based on zhuodu doctrine.Methods: 92 patients of CAG with intestinal metaplasia were collected from the Medical Department of Digestion of TCM Hospital of Hebei Province. They were randomly divided into treatment group and control group.Treatment group were given Hua Zhuo Jie Du decoction twice a day and one pouch in once for treatment, but control group were given Wei Fu Chun three times a day and four pills in once for treatment. One course of treatment was 12 weeks,two course of treatment in all.Results:1 The compairson of clinical therapeutic effect between two groups:A total of 52 cases of treatment group, 17 cases of clinical recovery, 21 cases of clinical markedly, 11 cases of effective, 3 cases of invalid, the total effective rate was 94.23%. A total of 40 cases of control group, 7 cases of clinical recovery, 10 cases of clinical markedly, 8 cases of effective, 15 cases of invalid, the total effective rate was 62.5%. The difference of clinical curative effect between two groups was significant （P<0.01）.2 The compairson of cardinal symptom score between two groups: After treating, the score of stomach pain, swelling of the liver is full, belching and debilitation decreased obviously in two groups.There was a significant difference bewteen before and after treating （treatment group and:1.32±0.51VS 4.82±1.25 P <0.01; control group:1.58±0.65 VS 4.69±1.17, P <0.01）. After treatment, treatment group in the above-mentioned symptoms was significantly lower than the control group, there was a significant difference （1.32±0.51 VS 1.58±0.65, P<0.05）.3 The compairson of gastroscopic effect between two groups: Gastroscopic curative effect of treatment group was superior to control group （The total effective rate in treatment group were 86.96%, 92%, 64.58%, 71.42%, 65.63%,the total effective rate control group were 60%, 78.95%, 38.23%, 36.36%, 31.82%, P<0.05 or P<0.01）. Treatment group and control group comparisons to improve the mucosal oedema was no significant difference（The total effective rate in treatment group were 87.5%, the total effective rate control group were71.88%, P>0.05）4 The compairson of Gastic mucosa’pathological curative effect between two groups:The total effective rates of Treatment group in the improvement of gastric atrophy and IM, were 78.85%, 84.62%. The total effective rate control group were 52.5%, 55%. Gastic mucosa’pathological curative effect of treatment group was superior to control group（P<0.01）.5 The compairson of the teradication rate of Hp between two groups: Two groups’curative effect of resising Hp infection were 47.22% and 44.44%. Two groups’curative effect of resisting Hp infection had no significant difference （P>0.05） .6 Detection of security indicators:There is no abnormal changes for Observation of blood, urine, conventional liver and kidney function and ECG changes before and after.Conclusions:1 Huazhuo Jiedu decoction can improve clinical symptoms in patients with CAG.2 Huazhuo Jiedu decoction can recover the gastric mucosa and improve intestinal metaplasia.3 Huazhuo Jiedu decoction can surely and partially resist Hp infection.Part 4:The effect on multiplication and apoptosis of human gastric cancer cell with drug-containing serum of Huazhuo Jiedu decoction and separate decoctionObjective: We observed the effect of multiplication and apoptosis to BGC-823 with drug-containing serum of Huazhuo Jiedu decoction and separate decoction in the experiment. The aim of it was to investigate if the Huazhuo Jiedu decoction could prevent precancerosis of CAG by the mechanism of action of inhibiting the cell to multiply and provide experimental evidence for clinical application.Methods:1 The preparation of drug-containing serum: 20 healthy male SD rats were randomly divided into four groups: normal group, Huazhuo Jiedu decoction group, Huazhuo decoction group, Jiedu decoction group.The amount of medicinal herb given to rats in Huazhuo Jiedu decoction group, Huazhuo decoction group, Jiedu decoction group was 345.9g·kg^-1·d^-1,208.4g·kg^-1·d^-1,137.3g·kg^-1·d^-1 weight respectively. Normal group were given 2ml isotonic Na chloride once a day by intragastric administration for 3 days. After anesthetizing by intraperitoneal injection hydral（350mg/kg）, all rats were killed and draw blood by abdominal aorta. After standing, centrifugd for 10min, 3000rpm/min, deactivated by 56℃for 30min, finally, conserved in frigidaire -20℃for pre-emergency.2 Administration and subgroup: we treated BGC-823 cell with blank pastilleserum, Huazhuo Jiedu drug-containing serum, Huazhuo decoction drug-containing serum, jiedu drug-containing serum. We departed them into four groups, Huazhuo Jiedu decoction group, Huazhuo decoction group, Jiedu decoction group and control group.3 Detected the index: we deployed MTT, flow cytometry, cell immunohistochemistry to detect the growth inhibiting, cell cycle, apoptosis rate, the expression of Bcl-2 and Bax proteinum in BGC-823 cell with different drug-containing serum.Results:1 The experiment on inhibitation the multiplication of cell: Compared with control group, there was effect on inhibitation the multiplication of BGC-823 cell with Huazhuo Jiedu decoction, Huazhuo decoction and Jiedu decoction （P<0.05,P<0.01）, especially, the effect of Huazhuo Jiedu decoction was evident, the rate of inhititation was 39.3%, 16.8%, 16.5% respectively. There was no different between Huazhuo decoction and Jiedu decoction in the effect of inhibitation.2 cell cycle: compared with normal group, great changes had happened in the cell cycle of BGC-823 with Huazhuo Jiedu decoction. The number of cell in G0/G1 stage increased （P<0.01）, the number of cell in S stage decreased（P<0.01）,there was no changes in G2/M stage, most of cells was hold in G0/G1 stage. But there was no changes in the cell cycle of BGC-823 with Huazhuo decoction and Jiedu decoction （P>0.05）.3 Apotosis rate: the BGC-823 was treated with different Drug-containing serum, the bar chart of DNA emerged typical hypodiploid apoptotic peak.The result of FCM: the apotosis rate in Huazhuo Jiedu group prescription,Huazhuo decoction group and Jiedu decoction group was （16.85±0.63）%, （11.48±0.50）%, （12.38±0.60）% respectively.Compared with normal group （2.09±0.25）%, there was significant difference between them （P <0.01）.There was significant difference between Huazhuo Jiedu decoction group, Huazhuo decoction group and Jiedu decoction group in apotosis rate （P <0.01）. There was no difference, Huazhuo decoction group and Jiedu decoction group in apotosis rate （P>0.05）.4 Detected the expression of Bcl-2 and Bax proteinum by cell immunohistochemistry: the masculine rate of expression Bcl-2 proteinum in huazhuojiedu group prescription, Huazhuo decoction group and Jiedu decoction group decreased greatly and they were （29.73±4.34）%, （37.11±4.63）%, （41.63±6.47）% respectively. Compared with normal group （53.74±2.97）%, there was significant statistical difference （P<0.01）. There was significant difference between Huazhuo Jiedu decoction group,Huazhuo decoction group and Jiedu decoction group in the masculine rate of expression Bcl-2 proteinum （P<0.01）. But there was no difference between Huazhuo decoction group and Jiedu decoction group （P>0.05）. The masculine rate of expression Bax proteinum in huazhuojiedu group prescription, Huazhuo decoction group and Jiedu decoction group was （58.11±4.19）%, （32.25±2.93）%, （34.84±2.66）% respectively. Compared with normal group （25.17±2.36）, there was significant difference （P<0.01）. There was significant difference between Huazhuo Jiedu decoction group, Huazhuo decoction group and Jiedu decoction group in the masculine rate of expression Bax proteinum （P<0.01）. But There was no difference between Huazhuo decoction group and Jiedu decoction group （P>0.05）.5 Detected the expression of Bcl-2 and Bax proteinum with flow cytometry:Compared with normal group the expression of Bcl-2 in Huazhuo Jiedu group prescription,Huazhuo decoction group and Jiedu decoction group decreased （257.67±9.02 VS 330.22±7.13 P<0.01; 291.22±11.07 VS 330.22±7.13 P<0.01;276.20±9.10VS330.22±7.13 P<0.01）.But the expression of Bax proteinum increased （330.14±8.19 VS 281.23±12.71 P<0.01; 303.84±6.60 VS 281.23±12.71P<0.05; 310.54±13.29 VS 281.23±12.71, P<0.01）.There was significant difference between Huazhuo Jiedu decoction group, Huazhuo decoction group and Jiedu decoction group in the expression of Bcl-2 and Bax proteinum （P<0.01）. There was no difference between Huazhuo decoction group and Jiedu decoction group in the expression of Bcl-2 and Bax proteinum（P>0.05）.Conclusions:1 Huazhuo Jiedu decoction can inhibit BGC-823 cell multiplication, because it can interact cell cycle and induce apoptosis.2 Huazhuo Jiedu decoction, Hua Zhuo decoction and Jie Du decoction induce apoptosis may be according to increase the expression of Bcl-2 proteinum and decrease the expression of Bax proteinum in BGC-823 cell.3 Huazhuo decoction and Jiedu decoction can induce apoptosis, but it can not influence the cell cycle. The compatibility of Huazhuo Jiedu decoction can retard cell in G0/G1 stage, therefore it has remarkable effect on inhibiting the BGC-823 cell multiplication of gastric adenocarcinoma.Conclusions:1 Our study investigated the problem about the type of syndrome in TCM and indicated that Zhuodu Zheng is one common type of syndrome of CAG by the the way of cluster analysis.2 The level of gastrin and motilin in blood plasma have variability in different type of syndrome of traditional chinese medicine of CAG. The change of the level of gastrin and motilin in blood plasma may be the reason to induce the different clinical manifestation among different type of syndrome.3 Huazhuo Jiedu decoction can improve clinical symptoms and intestinal metaplasia , recove gastric mucosa in CAG patients.4 The drug-containing serum of Huazhuo Jiedu decoction and separate decoction can inhibit BGC-823 cell to multiply and enhance to apoptosis. It is possible the one of mechanism of this prescription prevented gastric cancer and reversed precancerosis.更多还原