本文選題：雙胞胎 + 齲病��； 參考：《武漢大學(xué)》2017年碩士論文

【摘要】：齲病是學(xué)齡前兒童口腔內(nèi)最常見的疾病之一。低齡兒童齲(EarlyChildhood Caries,ECC)是指71個(gè)月或更小的兒童口腔中任意一個(gè)乳牙存在一個(gè)或一個(gè)以上齲壞(包括已形成齲洞及白堊色齲損)、因齲充填,因齲缺失牙齒。全國第三次口腔流行病學(xué)調(diào)查結(jié)果表明,乳牙患齲率在5歲兒童高達(dá)66%,顯著高于其他國家,表明在我國乳牙齲仍然是兒童口腔的一種常見病、多發(fā)病,現(xiàn)已成為一個(gè)重要的公眾衛(wèi)生問題,對(duì)兒童身體健康和生長發(fā)育都會(huì)產(chǎn)生較大的威脅,因而對(duì)齲病病因?qū)W進(jìn)行深入細(xì)致地研究,最終找到合理的預(yù)防措施的任務(wù)仍相當(dāng)艱巨。齲病病因的四聯(lián)因素理論認(rèn)為宿主、飲食、口腔中的微生物和時(shí)間共同參與了齲病的發(fā)生和發(fā)展,其中口腔微生物作為最重要的一個(gè)因素得到了口腔臨床醫(yī)生和科研工作者的廣泛關(guān)注,雖然有很多研究關(guān)注宿主的全身健康、飲食習(xí)慣、環(huán)境因素等對(duì)齲病發(fā)生的作用,但更多的還是關(guān)注口腔中可培養(yǎng)的微生物諸如變形鏈球菌、乳桿菌、放線菌等在引起齲病發(fā)生過程中的作用,家庭環(huán)境和遺傳因素對(duì)齲病發(fā)生的影響到目前為止還不是很明確。目的:本實(shí)驗(yàn)采用第二代人類口腔微生物芯片(Human Oral Microbe Identification using Next Generation Sequencing,HOMINGS)分析比較擁有不同患齲狀態(tài)雙胞胎兒童及其母親口腔內(nèi)齦上菌斑中菌群的結(jié)構(gòu)及微生物多樣性,從口腔細(xì)菌微生物層面和宿主遺傳及環(huán)境因素方面初步探討ECC的發(fā)病機(jī)制,為后期兒童齲病的預(yù)防、診斷與治療提供一定的理論依據(jù)。方法:根據(jù)世界衛(wèi)生組織(WHO)推薦的第4版齲病診斷標(biāo)準(zhǔn),對(duì)符合本本實(shí)驗(yàn)條件的武漢市3-6歲雙胞胎兒童及其母親進(jìn)行口腔健康檢查,要求所有受試者無系統(tǒng)性全身性疾病,口腔健康檢查前3個(gè)月內(nèi)未服用過抗生素,無中度或重度牙周疾病及嚴(yán)重口腔黏膜疾病。選取符合條件14對(duì)雙胞胎兒童(1人患齲1人無齲),1組三胞胎(2人患齲1人無齲)和15位母親,要求有齲組(n=16)DMFIT≥2,無齲組(n=15)DMFT=0。牙菌斑樣本取自14對(duì)雙胞胎、1組三胞胎兒童(年齡3-6歲)和15位母親,共46份樣本,分別提取齦上菌斑細(xì)菌總DNA,送到福賽研究中心,采用HOMINGS分析齦上菌斑中菌群結(jié)構(gòu)及微生物的多樣性。結(jié)果:1.通過HOMINGS所使用的Miseq測(cè)序平臺(tái)進(jìn)行高通量測(cè)序共獲得2,294,650條有效序列,使用QIIME軟件在相似性大于97%的水平上聚類操作分類單元(Operational taxonomic unit,OTU),共得到 20,431 個(gè) OTU,與 Greengene數(shù)據(jù)庫中的參考序列進(jìn)行比對(duì)和物種注釋獲得11個(gè)門,116個(gè)屬和189個(gè)種水平的物種;2.在門水平上,母親組在擬桿菌門和厚壁菌門的豐度顯著高于有齲組和無齲組;3.在屬水平上有齲組和無齲組比較,有齲組在鏈球菌屬、乳桿菌屬、放線菌屬、嗜血桿菌的豐度要高于無齲組(P0.05);母親組的微生物種類較兒童組多,分別在普氏菌屬、坦納氏菌屬的相對(duì)豐度高于兒童組(P0.05);4.Alpha多樣性分析結(jié)果表明母親組齦上菌斑微生物組成的豐富程度和均勻程度顯著高于有齲組和無齲組(P0.05);5.通過比較同卵和異卵雙胞胎之間加權(quán)的進(jìn)化距離(weighted unifrac distance),得到同卵雙胞胎較異卵雙胞胎齦上菌斑微生物組成更為相似。結(jié)論:1.鏈球菌屬、乳桿菌屬、放線菌屬、嗜血桿菌屬是與低齡兒童齲(ECC)顯著相關(guān)的"核心微生物組";2.成年人口腔內(nèi)齦上菌斑的微生物組成的豐富程度和均勻程度與兒童相比存在顯著差異,說明在不同年齡階段,口腔微生態(tài)環(huán)境是在發(fā)生變化的,而且成年人口腔內(nèi)的微生物菌群結(jié)構(gòu)使其更易患牙周疾病;3.同卵雙胞胎較異卵雙胞胎齦上菌斑的菌群組成更為相似說明遺傳因素對(duì)口腔微生態(tài)的物種組成是有一定影響的。
[Abstract]:Dental caries are one of the most common oral diseases in preschool children. EarlyChildhood Caries (ECC) refers to the existence of one or more caries (including caries and Cretaceous caries) in children's oral cavity of 71 months or younger children. The third oral epidemiology of dental caries is the result of caries filling and dental caries. The results show that the dental caries rate of 5 year old children is up to 66%, which is significantly higher than that of other countries. It shows that dental caries in our country are still a common disease in children's mouth. It has become an important public health problem, and has a great threat to the health and growth of children, so the caries etiology is deep. In detail, the task of finding reasonable preventive measures is still quite arduous. The quadruple factor theory of the cause of caries considers that the host, diet, microorganism and time in the oral cavity participate in the occurrence and development of dental caries. Oral microorganism is one of the most important factors in oral clinicians and researchers. Widely concerned, although many studies have focused on the effects of host body health, dietary habits and environmental factors on dental caries, more attention is paid to the role of cultured microbes in oral cavity, such as Streptococcus mutans, Lactobacillus, actinomycetes, etc., in the process of dental caries, family environment and genetic factors. The effect is not clear so far. Objective: To compare the structure and microbial diversity of the bacteria in the oral gingival plaque of the twin children with different caries status and their mothers with second generations of Human Oral Microbe Identification using Next Generation Sequencing, HOMINGS. The pathogenesis of ECC is preliminarily discussed from the aspects of oral bacterial microorganism and host genetic and environmental factors, which provide a certain theoretical basis for the prevention, diagnosis and treatment of children's caries in the later period. Methods: according to the fourth version of the dental caries recommended by the WHO (WHO), the 3-6 year old double cell fetus in Wuhan City, which is in accordance with the original experimental conditions Children and their mothers were examined for oral health, requiring all subjects to have no systemic systemic disease, no antibiotics in the first 3 months of oral health examination, no moderate or severe periodontal disease and severe oral mucosal disease. 14 pairs of twins (1 with caries, 1 without caries), and 1 triplets (2 caries 1 without caries) were selected. 15 mothers were required to have a caries group (n=16) DMFIT > 2. The caries free group (n=15) DMFT=0. plaque samples were taken from 14 pairs of twins, 1 triplet children (age 3-6 years old) and 15 mothers, with 46 samples, respectively, to extract the total DNA of the bacteria on the gingival plaque, sent to the fusai Research Center, and used HOMINGS to analyze the flora structure and microbial diversity in the supra gingival plaque. Results: 1. a total of 2294650 effective sequences were obtained by high flux sequencing using the Miseq sequencing platform used by HOMINGS. Using QIIME software, a total of 20431 OTU, which were compared with the reference sequence in the Greengene database, was compared with the reference sequence in the Greengene database, and the Operational taxonomic unit, OTU was used in the level of similarity greater than 97%. The annotation obtained 11 gates, 116 genera and 189 species of species; 2. at the portal level, the abundance of the mother group in the bacteriobacterium and the thick wall bacteria was significantly higher than that in the caries and the caries free groups. 3. at the level of caries and non caries, the abundances of the caries, the lactobacillus, the actinomycetes, and the actinomycetes in the caries group were higher than those in the caries free group (P0.05 The microbial species in the mother group were more than those in the children's group, and the relative abundance of the genus tanabi was higher than that in the children's group (P0.05). The 4.Alpha diversity analysis showed that the abundance and uniformity of the microbial composition of the gingival plaque in the mother group was significantly higher than that in the caries and the caries free groups (P0.05); 5. by comparing the identical and heterogenous twins. The weighted evolutionary distance (weighted UniFrac distance) showed that identical twins were more similar to the microbiological composition of the supra gingival plaque in identical twins. Conclusion: 1. Streptococcus, Lactobacillus, actinomycetes, and Haemophilus is a "core microorganism group" that is significantly associated with low age children's caries (ECC); 2. adult oral gingival plaque The richness and uniformity of biological composition are significantly different from those of children, indicating that the microecological environment of the oral cavity is changing in different ages, and the microbial flora in the oral cavity of adults makes it more susceptible to periodontal disease, and 3. identical twins are more similar than those of the gingival plaque of the ISO twins. It indicates that genetic factors have certain effects on the species composition of oral microecology.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R788.1

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