本文選題：益生菌 + 潰瘍性結(jié)腸炎癌變　； 參考：《北京協(xié)和醫(yī)學(xué)院》2017年博士論文

【摘要】：第一部分益生菌對(duì)潰瘍性結(jié)腸炎癌變的抑制作用及腸道菌群分析目的:分析益生菌VSL#3、布拉氏酵母菌干預(yù)對(duì)AOM/DSS誘導(dǎo)的潰瘍性結(jié)腸炎癌變模型小鼠成瘤的影響,同時(shí)分析小鼠糞便和黏膜菌群的變化,探討腸道菌群變化在UC癌變中的作用。方法:C57BL/6小鼠隨機(jī)分為7組。第1-6組建立潰瘍性結(jié)腸炎癌變模型,第1周第1天予腹腔注射12.5mg/kg AOM,1周后開始連續(xù)5天予含2.5%DSS的飲用水,隨后更換成普通飲用水。其中第1-5組為不同干預(yù)組,即從注射AOM當(dāng)日起分別予 5-ASA(75mg/kg/d)、布拉氏酵母菌(5×107CFU/d)、VSL#3(1.5×l09CFU/d)、5-ASA聯(lián)合布拉氏酵母菌、5-ASA聯(lián)合VSL#3灌胃處理(5d/周)。第6組為模型對(duì)照組,不予灌胃干預(yù)。第7組為空白對(duì)照組,不予建模及灌胃干預(yù)。第12周末處死小鼠,統(tǒng)計(jì)比較各組瘤負(fù)荷。收集試驗(yàn)前后小鼠糞便及試驗(yàn)后腸黏膜標(biāo)本,通過16s rDNA擴(kuò)增子測(cè)序方法分析各組腸道菌群差異。結(jié)果:1.各組小鼠成瘤率:干預(yù)組63-85%,模型對(duì)照組100%。2.各組小鼠瘤負(fù)荷:5-ASA 組(0.43±0.14cm)、布拉氏酵母菌組(0.20±0.07cm)、VSL#3 組(0.25±0.07cm)、5-ASA+布拉氏酵母菌組(0.53±0.1lcm)、5-ASA+VSL#3 組(0.46±0.11cm)瘤負(fù)荷較模型對(duì)照組(0.97±0.19cm)均顯著下降,差異有統(tǒng)計(jì)學(xué)意義(p=0.0269,p=0.0006,p=0.0009,p=0.0486,p=0.0261)。3.糞便菌群分析顯示:(1)模型對(duì)照組較空白對(duì)照組乳桿菌屬減少,顫桿菌屬、Lachnoclostridium增加。(2)益生菌干預(yù)后,芽孢桿菌屬、乳球菌屬、乳桿菌屬增加,顫桿菌屬、Lachnoclostridium、擬桿菌屬、假單胞菌屬減少。4.腸道黏膜菌群分析顯示:(1)模型對(duì)照組較空白對(duì)照組瘤胃球菌科014菌屬、雙歧桿菌屬減少,擬普雷沃菌屬增加。(2)較模型對(duì)照組相比,布拉氏酵母菌組毛螺旋菌科NK4A136菌屬、Lachnoclostridium增加;VSL#3組雙歧桿菌屬增加。結(jié)論:l.VSL#3和布拉氏酵母菌能夠顯著降低UC癌變小鼠的成瘤率和瘤負(fù)荷。2.UC癌變小鼠腸道菌群紊亂,黏膜和糞便菌群均表現(xiàn)為有益菌屬減少、有害菌屬增加。3.補(bǔ)充益生菌有益于UC癌變小鼠糞便和黏膜菌群實(shí)現(xiàn)再平衡。4.雙歧桿菌屬容易在腸黏膜定植,芽孢桿菌屬和乳球菌屬容易在糞便定植。第二部分炎癥因子及布拉氏酵母菌對(duì)Wnt信號(hào)通路的影響及機(jī)制研究目的:探討炎癥因子IL-6、TNF-α及布拉氏酵母菌對(duì)Wnt信號(hào)通路的影響及作用機(jī)制。方法:選用正常腸黏膜上皮細(xì)胞CCC-HIE-2、結(jié)腸癌細(xì)胞Caco-2和HCT-116,分別予IL-6和TNF-α刺激,采用雙熒光素酶報(bào)告基因?qū)嶒?yàn)檢測(cè)Wnt信號(hào)通路活性;給予NF-(?)B信號(hào)通路特異性抑制劑PDTC,觀察PDTC對(duì)IL-6和TNF-α調(diào)節(jié)Wnt信號(hào)通路的影響;將布拉氏酵母菌與細(xì)胞共培養(yǎng),觀察對(duì)IL-6和TNF-α調(diào)節(jié)Wnt信號(hào)通路的影響。結(jié)果:1.不同濃度IL-6和TNF-α刺激CCC-HIE-2、Caco-2、HCT-116均能使相對(duì)熒光素酶活性升高,上調(diào)Wnt信號(hào)通路。IL-6和TNF-α刺激細(xì)胞的最佳濃度及時(shí)間均為 50ng/ml,6h。2.PDTC 可以抑制 IL-6 和 TNF-α 對(duì) CCC-HIE-2 細(xì)胞(1.044+0.006 vs 0.895±0.003 p=0.0032,1.126±0.006 vs l.062±0.014/p=0.0006)、Caco-2 細(xì)胞(8.859±0.050 vs 7.922±0.075 p=0.0046,3.951±0.006 vs 3.550±0.022 p=0.0016)和 HCT-116 細(xì)胞(2.728±0.053 vs2.430+0.022p=0.0008,4.044±0.022 vs 3.629±0.025p=0.0033)的上調(diào)相對(duì)熒光素酶活性作用。3.布拉氏酵母菌可以抑制 IL-6 和 TNF-α 對(duì) CCC-HIE-2 細(xì)胞(1.239±0.015 vs 0.956±0.001 p=0.0014,1.022±0.014 vs 0.780±0.026 p=0.0074)、Caco-2 細(xì)胞(9.731±0.075 vs 8.479±0.099 p=0.0049,9.187±0.044vs7.691±0.055p=0.0011)和 HCT-116 細(xì)胞(4.116±0.004 vs3.734±0.045p=0.0068,4.489±0.010vs 3.858±0.021p=0.0007)的上調(diào)相對(duì)熒光素酶活性作用。結(jié)論:1.在 CCC-HIE-2、Caco-2、HCT-116 細(xì)胞系,TNF-α 和 1L-6 能夠通過 NF-(?)B通路上調(diào)Wnt信號(hào)通路。2.布拉氏酵母菌與CCC-HIE-2、Caco-2、HCT-116細(xì)胞共培養(yǎng),能夠抑制TNF-α和IL-6對(duì)Wnt信號(hào)通路的上調(diào)。第三部分不同病變范圍潰瘍性結(jié)腸炎患者腸道菌群分析目的:探討不同病變范圍潰瘍性結(jié)腸炎患者腸道菌群特點(diǎn)。方法:收集10例健康體檢個(gè)體糞便標(biāo)本,35例UC患者(直腸型7例,左半結(jié)腸型9例,廣泛結(jié)腸型19例)糞便及左半、右半腸黏膜標(biāo)本,通過16srDNA擴(kuò)增子測(cè)序方法分析腸道菌群差異。結(jié)果:1.糞便菌群分析顯示:(1)廣泛結(jié)腸型UC患者較健康個(gè)體菌群多樣性顯著減少。(2)較健康個(gè)體相比,UC患者Subdoligranulum、瘤胃球菌屬-2、Fusicatenibacter、柔嫩梭菌屬、假丁酸弧菌屬、Alistipes減少;埃希氏菌屬、Peptoclostridium屬及韋榮氏菌屬增加。(3)直腸型UC較左半結(jié)腸型UC患者糞便菌群假丁酸弧菌屬增加,較廣泛結(jié)腸型UC患者毛螺旋菌屬增加、普雷沃氏菌屬-9減少。2.黏膜菌群分析表明:(1)較直腸型患者相比,左半結(jié)腸型和廣泛結(jié)腸型患者乳球菌屬、Geobacillus、鏈球菌屬、明串珠菌屬減少。(2)較廣泛結(jié)腸型患者相比,直腸型和左半結(jié)腸型患者擬桿菌屬減少。(3)所有類型UC患者左半與右半黏膜菌群生物多樣性及菌屬組成無統(tǒng)計(jì)學(xué)差異。(4)UC患者腸道黏膜較糞便菌群生物多樣性增多,菌屬組成存在差異。結(jié)論:1.UC患者糞便菌群生物多樣性降低,有益菌屬減少,有害菌屬增加。2.隨著病變范圍增大,患者糞便和腸黏膜菌群均表現(xiàn)為有益菌屬減少,有害菌屬增加。3.患者黏膜菌群的改變呈全結(jié)腸分布改變特點(diǎn),不局限于病變部位黏膜。4.患者糞便和黏膜菌群生物多樣性及菌屬組成存在差異。目的:探討胃神經(jīng)內(nèi)分泌腫瘤(Gastric Neuroendocrine Neoplasms,GNENs)的臨床特征,以提高臨床醫(yī)師對(duì)該病的認(rèn)識(shí)。方法:收集2005年10月至2015年10月北京協(xié)和醫(yī)院收治的36例GNENs,對(duì)其人口學(xué)特征、主要臨床表現(xiàn)、內(nèi)鏡下表現(xiàn)、病理特征及治療情況進(jìn)行回顧性分析。結(jié)果:1.36例患者平均年齡55.8歲,男女比例0.89:1。2.臨床表現(xiàn)多種多樣,腹痛的發(fā)生率高達(dá)42.9%。3.63.9%的病變分布于胃體部。內(nèi)鏡下病變可表現(xiàn)為息肉樣隆起、潰瘍性病變、黏膜凹陷。以息肉樣隆起最為多見,其中73.9%為單發(fā)病變,平均直徑小于l0mm。4.病理分級(jí)包括G1-G3級(jí),以G1級(jí)居多(55.6%)。5.65.2%于胃體行活檢的患者病理證實(shí)存在胃體黏膜萎縮或腸化。6.治療方案主要包括內(nèi)鏡和手術(shù)根治。結(jié)論:1.GNENs缺乏特異性臨床表現(xiàn),病變以胃體分布居多,在內(nèi)鏡下多表現(xiàn)為息肉樣隆起。2.對(duì)于合并萎縮性胃炎,尤其胃體萎縮的息肉樣病變,應(yīng)警惕GNENs的可能性。
[Abstract]:The first part of probiotics inhibit carcinogenesis of ulcerative colitis and intestinal microflora analysis objective: analysis of probiotics VSL#3 intervention ofsaccharomyces boulardii tumor effect on mice ulcerative colitis carcinogenesis model induced by AOM/DSS, and the analysis of mice feces and mucosal flora changes, to explore the changes of intestinal microflora in UC carcinogenesis. Methods C57BL/6 mice were randomly divided into 7 groups. The 1-6 group to establish ulcerative colitis carcinogenesis model, first weeks and first days by intraperitoneal injection of 12.5mg/kg AOM, 1 weeks after the start of 5 consecutive days to drinking water containing 2.5%DSS, then replace the ordinary drinking water. The 1-5 group is different from the intervention group, the injection of AOM since they were given 5-ASA (75mg/kg/d), Saccharomyces boulardii (5 * 107CFU/d), VSL#3 (1.5 * l09CFU/d), 5-ASA and 5-ASA combined with VSL#3 boulardii, gavage treatment (5d/ weeks). The sixth groups as model control group, not Stomach intervention. The seventh group was the control group, no modeling and stomach intervention. At the end of the twelfth mice were killed, were collected. Statistical tests before and after the tumor burden mice feces and test of intestinal mucosa specimens by 16S rDNA amplicon sequencing method to analyze the difference of intestinal flora. Results: 1. groups of mice: the intervention rate group 63-85%, model group 100%.2. mice tumor burden: group 5-ASA (0.43 + 0.14cm), Saccharomyces boulardii group (0.20 + 0.07cm), group VSL#3 (0.25 + 0.07cm), 5-ASA+ group of Saccharomyces boulardii (0.53 + 0.1lcm), group 5-ASA+VSL#3 (0.46 + 0.11cm) tumor burden compared with model control group (0.97 + 0.19cm) were significantly decreased, the difference was statistically significant (p=0.0269, p=0.0006, p=0.0009, p=0.0486, p=0.0261).3. fecal flora analysis showed that: (1) the model group compared with blank control group Lactobacillus decreased, Lachnoclostridium increased. AF bacillus, probiotics (2) After the intervention, bacillus, Lactococcus, Lactobacillus, Bacillus Lachnoclostridium, fibrillation, Bacteroides spp., Pseudomonas.4. reduce the intestinal mucosal flora analysis showed that: (1) model group compared with blank control group, 014 Ruminococcus genus, the genus Bifidobacterium reduced to Prevotella sp. is increased. (2) compared with the model group compared with the control group, Saccharomyces boulardii lachnospiraceae, NK4A136 sp., Lachnoclostridium increased in VSL#3 group; Bifidobacterium increased. Conclusion: l.VSL#3 and Saccharomyces boulardii can significantly reduce the rate of tumor and tumor load.2.UC cancer mice intestinal flora disorder UC cancer mice, mucosa and fecal bacteria the group showed good bacteria is reduced, harmful bacteria increased.3. probiotics is beneficial to UC cancer mice feces and mucosal flora to rebalance.4. Bifidobacterium easily in intestinal colonization, bacillus and Lactococcus. In the fecal colonization. Objective to study the effect of Wnt signal pathway and mechanism of the second part: To investigate the inflammatory factors and Saccharomyces boulardii inflammatory cytokine IL-6, TNF- alpha and Saccharomyces boulardii effect on Wnt signaling pathway and the mechanism. Methods: the CCC-HIE-2 of normal intestinal epithelial cells, Caco-2 and HCT-116 colon cancer cells were treated with IL-6 and TNF- alpha stimulation by Wnt signaling pathway activated luciferase reporter gene experiment; NF- (?) B pathway inhibitor PDTC, observe the PDTC Wnt signal pathway in regulating IL-6 and TNF- alpha will influence; Saccharomyces boulardii and cell co culture, observe the effect on IL-6 and TNF- alpha Wnt pathway results: 1. different concentrations of IL-6 and TNF- CCC-HIE-2 Caco-2, HCT-116 alpha stimulation, can make the relative luciferase activity increased, the optimum concentration of upregulation of Wnt signaling pathway of.IL-6 and TNF- stimulated cells And the time of 50ng/ml, IL-6 and 6h.2.PDTC can inhibit TNF- alpha on CCC-HIE-2 cells (1.044+0.006 vs 0.895 + 0.003 p=0.0032,1.126 + 0.006 vs l.062 + 0.014/p=0.0006), Caco-2 cells (8.859 + 0.050 vs 7.922 + 0.075 p=0.0046,3.951 + 0.006 vs 3.550 + 0.022 p=0.0016) and HCT-116 cells (2.728 + 0.053 vs2.430+0.022p=0.0008,4.044 + 0.022 vs 3.629 + 0.025p=0.0033 the relative luciferase activity) by.3. boulardii inhibits IL-6 and TNF- alpha on CCC-HIE-2 cells (1.239 + 0.015 vs 0.956 + 0.001 p=0.0014,1.022 + 0.014 vs 0.780 + 0.026 p=0.0074), Caco-2 cells (9.731 + 0.075 vs 8.479 + 0.099 p=0.0049,9.187 + 0.044vs7.691 + 0.055p=0.0011) and HCT-116 cells (4.116 + 0.004 + 0.045p=0.0068,4.489 + vs3.734 0.010vs 3.858 + 0.021p=0.0007) increased relative luciferase activity effect. On: 1. in CCC-HIE-2, Caco-2, HCT-116 cell lines, TNF- alpha and 1L-6 by NF- (?) B pathway. Wnt pathway.2. boulardii with CCC-HIE-2, Caco-2, HCT-116 cells were co cultured, can inhibit TNF- alpha and IL-6 on the up regulation of the Wnt signaling pathway. The third part is the same lesion in patients with ulcerative colitis the intestinal flora analysis objective: To investigate the different extent of disease in patients with ulcerative colitis intestinal flora characteristics. Methods: We collected 10 cases of healthy individual fecal samples from 35 UC patients (rectum type in 7 cases, 9 cases of left colon type extensive colonic type 19 cases) stool and left half, right through the intestinal mucosa specimens. 16srDNA amplicon sequencing method difference analysis of intestinal flora. Results: 1. fecal flora analysis showed that: (1) patients with extensive colon type UC compared with healthy individuals flora diversity decreased significantly. (2) compared with healthy individuals, UC patients with Subdoligranulum, rumen ball Genus of -2, Fusicatenibacter, and Clostridium, false butyrivibrio, Alistipes decreased; Escherichia coli, genus Peptoclostridium and genus Veillonella. (3) UC with left colon rectal UC patients fecal flora false butyrivibrio increased widely colonic type UC patients lachnospiraceae genus Poulet Was added, genus -9 reduced.2. mucosal flora analysis showed that: (1) a type of rectal patients with left colon and colon were widely Lactococcus, Geobacillus, Streptococcus, Leuconostoc. (2) compared with patients with extensive colitis, rectum and left colon in patients with type bacteriodes decreased. (3) all types of UC patients with left-sided and right-sided mucosal flora biodiversity and species composition had no significant difference. (4) UC with intestinal mucosa of patients with fecal flora biodiversity increased, species composition is different. Conclusion: 1.UC patients with fecal flora biology Reduced diversity, beneficial bacteria to reduce harmful bacteria is increased with.2., the lesion increased, and stool of patients with intestinal mucosa flora showed beneficial bacteria is reduced, harmful bacteria increased.3. mucosal flora of patients with changes in total colonic distribution characteristics, not limited to the lesion mucosa and mucosal.4. in patients with fecal bacteria group of biological diversity and species composition differences. Objective: To investigate the gastric neuroendocrine tumor (Gastric Neuroendocrine, Neoplasms, GNENs) of the clinical features, to improve the understanding of this disease. Methods: 36 cases of GNENs from October 2005 to October 2015 in Peking Union Medical College Hospital were, on the demographic characteristics, main clinical manifestations, endoscopic findings. The pathological characteristics and treatment were analyzed retrospectively. Results: 1.36 cases of patients with an average age of 55.8 years, the proportion of male and female 0.89:1.2. variety of clinical manifestations, the high incidence of abdominal pain As 42.9%.3.63.9% lesions distributed in the body of stomach. Endoscopic lesions showed polypoid bulge, ulcer lesions, mucosal polypoid bulge sag. In the most common, of which 73.9% were single lesions, the average diameter of less than l0mm.4. pathological grades including G1-G3 level, G1 level in the majority (55.6%) of patients with pathological.5.65.2% in the stomach body biopsies confirmed the presence of.6. treatment of gastric mucosa atrophy or intestinal metaplasia including radical endoscopy and surgery. Conclusion: 1.GNENs is lack of specific clinical manifestations. The lesion in the gastric body distribution mostly in endoscopic manifestations of polypoid bulge.2. for patients with atrophic gastritis, especially polypoid lesions of gastric atrophy. Should be alert to the possibility of GNENs.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R574.62

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 葉燕明,鄧明輝,鄭文寬,馬燕,魏偉珩,陳國(guó)興,黎麗,劉定南;傳染性非典型肺炎7例臨床分析[J];中國(guó)熱帶醫(yī)學(xué);2004年02期

2 張衛(wèi)華;;特發(fā)性間質(zhì)性肺炎臨床治療體會(huì)[J];北方藥學(xué);2013年04期

3 宋延榮;程育春;徐勇全;;真性紅細(xì)胞增多癥1例報(bào)告[J];實(shí)用醫(yī)技雜志;2007年04期

4 韓萬柏,徐祖權(quán);卡氏肺孢子蟲肺炎[J];解放軍醫(yī)學(xué)高等專科學(xué)校學(xué)報(bào);1999年03期

5 魏曉華;;睡眠呼吸暫停綜合征臨床表現(xiàn)及治療探討[J];中國(guó)現(xiàn)代藥物應(yīng)用;2009年23期

6 王海宏;;急性病毒性心肌炎100例臨床探析[J];中國(guó)醫(yī)學(xué)裝備;2013年03期

7 李如堯,李寶林,張惠麗;特發(fā)性肺含鐵血黃素沉著癥9例臨床分析[J];臨床醫(yī)學(xué);2005年02期

8 李國(guó)愈;;甲狀腺功能亢進(jìn)癥的中醫(yī)藥治療體會(huì)[J];求醫(yī)問藥(下半月);2011年07期

9 唐金榮,侯熙德,王穎,李弘鈞,金慶文,丁新生;單純皰疹病毒性腦炎的臨床表現(xiàn)[J];卒中與神經(jīng)疾病;1999年02期

10 彭孝清;;老年急性呼吸窘迫綜合征的臨床表現(xiàn)及治療[J];醫(yī)學(xué)信息(上旬刊);2011年04期

相關(guān)會(huì)議論文 前1條

1 秦永新;萬獻(xiàn)堯;;血栓性血小板減少性紫癜相關(guān)危險(xiǎn)因素初探[A];中國(guó)重癥醫(yī)學(xué)的創(chuàng)新與發(fā)展——第二屆中國(guó)醫(yī)師協(xié)會(huì)重癥醫(yī)學(xué)醫(yī)師分會(huì)暨第九屆中國(guó)病理生理學(xué)會(huì)危重病醫(yī)學(xué)專業(yè)委員會(huì)2010年危重病醫(yī)學(xué)全國(guó)會(huì)議會(huì)刊[C];2010年

相關(guān)博士學(xué)位論文 前1條

1 王春賽爾;益生菌抑制潰瘍性結(jié)腸炎癌變的相關(guān)機(jī)制及腸道菌群差異分析[D];北京協(xié)和醫(yī)學(xué)院;2017年

相關(guān)碩士學(xué)位論文 前2條

1 張青青;子宮肌瘤發(fā)病相關(guān)危險(xiǎn)因素及手術(shù)方式的調(diào)查與分析[D];皖南醫(yī)學(xué)院;2015年

2 劉玲;兒童慢性免疫性血小板減少癥的臨床研究[D];重慶醫(yī)科大學(xué);2014年

，

本文編號(hào)：1760440