健康人和胃癌患者的舌苔厚度和菌群差异研究

发布时间：2018-05-31 02:41

本文选题：胃癌 + 舌诊　；参考：《皖南医学院》2016年硕士论文

【摘要】：目的:中医是我国的传统医学,具有悠久的历史丰富的临床使用经验。舌诊有无创和方便的优点,但因为其主观性较强且缺乏标准性,不能得到西方医学广泛认同。本研究的目的在于揭示舌苔厚度与舌苔菌群的关系,并尝试发现胃癌病人口腔菌群的变化,为舌诊和胃癌研究提供一个新方向。方法:我们利用病例对照研究的方法,用DS01-B舌苔信息分析系统去比较胃癌患者和健康人的舌苔厚度,并利用该软件将他们分为厚苔组和薄苔组;再分别刮取他们的舌苔标本,提取标本中总DNA,扩增其中16S r DNA的V2-V4高变区,利用高通量测序技术对胃癌患者和健康人舌苔中的16S r RNA基因进行测序分析;为了获得更精确的数据,我们筛选不合格的序列,利用QIIME软件将每个序列分类成一个OTU,将每个OTU与SILVA数据库比对以获得分类学数据;为了获得舌苔菌群多样性的数据,我们借助ACE、Chao和Shannon指数对不同的个体进行了α多样性进行了分析,并在纲、目、科、属、种水平对厚苔与薄苔之间、胃癌患者和健康人之间进行了菌群结构的分析。结果:我们分析比较了74个胃癌病人和72个健康人的舌苔厚度,发现胃癌组的舌苔厚度明显厚于健康对照组(343.11±198.22 vs.98.42±48.25,P0.001);在74个胃癌病人中有51.35%的个体拥有厚苔(38/72),而在正常人所有的个体都拥有薄苔;为了获得更好的标本用于高通量测序,我们挑选了18个拥有厚苔的胃癌患者舌苔标本、16个薄苔胃癌标本和16个健康人标本分成3组进行高通量测序;利用高通量测序技术我们在50个舌苔标本中共对1155532个序列512575267个碱基进行测序,通过与数据库的比对共鉴定出20个菌门、162个菌属和350个菌种;通过对3组的菌群α多样性进行比较,我们发现胃癌厚苔组的ACE,Chao和Shannon指数均明显低于胃癌薄苔组和健康对照组,而健康对照组和胃癌薄苔组的指数基本相同;在属的水平对胃癌薄苔和厚苔组菌群丰度进行分析时发现,胃癌薄苔组舌苔中丰度位于前五位的分别是:普氏菌属,韦永氏球菌属,纤毛菌属,乳球菌属和链球菌属,在胃癌厚苔组中位于前五位的分别是:普氏菌属,链球菌属,放线菌属,韦永氏球菌属和纤毛菌属。与薄苔组相比,厚苔胃癌患者舌苔拥有更高丰度的放线菌属和链球菌属。在种的水平对3组进行比较时发现,3组共同拥有的菌种有225种,有32种仅在健康人中发现,有47种仅在胃癌患者种发现,有17种仅在厚苔中发现;在对胃癌组和健康对照组在门水平进行比较它们的厚壁菌门、拟杆菌门、梭杆菌门和TM7相对丰度无明显差异,但在胃癌组中变形菌门的相对丰度明显小于对照组(10.85%vs.28.55%,P0.001)而放线菌明显多于对照组(12.32%vs.4.46%,P0.001)。健康人舌苔变形菌门相对丰度的平均值为28.55%(最低丰度11.70%,最高丰度42.99%);胃癌患者舌苔变形菌门相对丰度的平均值为10.85%(最低丰度0.11%,最高丰度34.77%).其中16个健康人中只有1人的变形菌门相对丰度低于15%,而在34个胃癌患者中有25人的变形菌门相对丰度低于15%。结论:(1)利用舌苔分析仪对舌苔进行分析时,胃癌病人的舌苔明显厚于健康人,约有一半的胃癌患者拥有厚苔;(2)舌苔厚度与菌群多样性相关,厚苔的菌群多样性明显低于薄苔;(3)和健康人相比,胃癌病人拥有不同的口腔菌群结构;
[Abstract]:Objective: Traditional Chinese medicine is a traditional medicine in China. It has a long history of clinical use. The tongue diagnosis has the advantages of non-invasive and convenient. But because of its strong subjectivity and lack of standard, it can not be widely recognized by western medicine. The purpose of this study is to reveal the relationship between the thickness of the tongue coating and the flora of the tongue coating, and try to find the patients with gastric cancer. The changes in oral flora provide a new direction for the study of tongue diagnosis and gastric cancer. Methods: we use a case-control study to compare the tongue coating thickness of gastric cancer patients and healthy people with DS01-B tongue coating information analysis system, and use the software to divide them into thick moss and thin moss groups, and then scrape their tongue coating specimens to extract the standard. In the total DNA, we amplify the V2-V4 high variable region of 16S R DNA and sequence analysis of 16S R RNA gene in gastric cancer patients and healthy people with high throughput sequencing technology. In order to obtain more accurate data, we select unqualified sequences and classify each sequence into a OTU with QIIME software, and compare each OTU to SILVA database. To obtain taxonomic data, in order to obtain the data of the diversity of the tongue coating flora, we analyzed the diversity of the different individuals with the help of ACE, Chao and Shannon index, and analyzed the structure of the flora between the thick moss and the thin moss, the gastric cancer patients and the healthy people in the class, order, family, genus, species. The tongue coating thickness of 74 gastric cancer patients and 72 healthy people was compared. It was found that the thickness of the tongue coating was thicker than that of the healthy control group (343.11 + 198.22 vs.98.42 + 48.25, P0.001), and 51.35% of the 74 gastric cancer patients had thick moss (38/72), and all of the individuals in the normal people had thin moss. We selected 18 tongue coating specimens of gastric cancer patients with thick moss, 16 thin moss gastric cancer specimens and 16 healthy human specimens, which were divided into 3 groups to carry out high flux sequencing. By using high throughput sequencing technology, we sequenced 1155532 sequences of 512575267 bases in 50 tongue coating specimens, and identified by comparison with the database. 20 bacteria doors, 162 bacteria and 350 strains; by comparing the diversity of the 3 groups of bacteria, we found that the ACE, Chao and Shannon indices of the thick gastric carcinoma group were significantly lower than those of the gastric carcinoma and healthy controls, while the index of the healthy control group and the gastric carcinoma group was basically the same; the level of the gastric carcinoma and the thick moss group were abundant at the level of the genus. When analyzed, it was found that the first five sites of the tongue fur of the gastric carcinoma group were: prytococcus, Wei Yong, cilium, Lactococcus and Streptococcus. The first five were in the thick moss of gastric cancer: the genus prytococcus, Streptococcus, actinomycetes, Micrococcus and cilium. Compared with the thin moss group, The tongue fur of thick mossy stomach cancer patients had higher abundance of actinomycetes and Streptococcus. When compared to the 3 groups, 225 species were found in the 3 groups, 32 were found only in healthy people, 47 were found only in gastric cancer, 17 were only in thick moss, and in the gastric cancer group and the healthy control group at the portal level. The relative abundances of the bacteriobacteria, Clostridium and TM7 were not significantly different, but the relative abundance of the Proteus doors in the gastric cancer group was significantly smaller than that of the control group (10.85%vs.28.55%, P0.001), and the actinomycetes were significantly more than the control group (12.32%vs.4.46%, P0.001). The average value of the relative abundance of the tongue fur deformable bacteria was 28.55% (12.32%vs.4.46%, P0.001). The minimum abundance of 11.70% and the highest abundance of 42.99%); the average value of the relative abundance of the tongue fur Proteus of gastric cancer patients was 10.85% (the lowest abundance 0.11%, the highest abundance of 34.77%). Only 1 of the 16 healthy people were less than 15% of the Proteus door relative abundance, and 25 of the 34 gastric cancer patients were compared with the 15%. conclusion: (1) utilization. When the tongue coating analyzer was used to analyze the tongue coating, the tongue fur of the gastric cancer patients was thicker than that of the healthy people, and about half of the gastric cancer patients had thick moss; (2) the thickness of the tongue fur was related to the diversity of the flora, and the diversity of the thick moss was significantly lower than that of the thin moss; (3) the gastric cancer patients had different oral flora structure compared with those of the healthy people.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2

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