本文選題：口腔扁平苔蘚　切入點：外周血淋巴細(xì)胞　出處：《南京醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探索口腔扁平苔蘚(Oral Lichen Planus, OLP)外周血淋巴細(xì)胞中游離鈣[Ca2+]i水平,及與之密切相關(guān)的兩種調(diào)控蛋白：Golli-MBP (Genes of the Oligodendrocyte lineage-Myelin Basic Protein,少突膠質(zhì)細(xì)胞譜系基因-髓鞘堿性蛋白)和STIM1(Stromal Interaction Molecule1,基質(zhì)相互作用分子1)的表達(dá)水平,探討Golli-MBP、STM1對口腔扁平苔蘚外周免疫細(xì)胞的影響,初步探索口腔扁平苔蘚的發(fā)病機制。 方法：本實驗采用鈣離子熒光探針Fluo-3檢測43例OLP患者(22例非糜爛型及21例糜爛型)外周血淋巴細(xì)胞和20例正常人外周血淋巴細(xì)胞中游離鈣離子水平。采用實時定量PCR(Quantitive Real-Time PCR, qPCR)和免疫蛋白印記技術(shù)(Western Blot),分別檢測43例OLP患者和20例正常人外周血淋巴細(xì)胞中Golli-MBP和STIM1的基因和蛋白表達(dá)水平。統(tǒng)計學(xué)分析各組數(shù)據(jù),比較各組指標(biāo)是否存在統(tǒng)計學(xué)差異。 結(jié)果：檢測結(jié)果表明,OLP患者外周血淋巴細(xì)胞受到活化刺激后,胞內(nèi)游離鈣離子升高幅度顯著低于正常對照組(P0.001)。Golli-MBP mRNA和蛋白在OLP組中的表達(dá)顯著高于正常對照組(P0.001)。同樣,STIM1mRNA和蛋白在OLP組中的表達(dá)亦顯著高于正常對照組(P0.01)。 結(jié)論：OLP患者外周血淋巴細(xì)胞胞內(nèi)游離鈣離子濃度在活化過程中升高水平低于對照組,提示OLP的淋巴細(xì)胞鈣離子信號通路可能存在異常,該通路涉及的外周血免疫細(xì)胞活化可能存在障礙。本實驗亦發(fā)現(xiàn)Golli-MBP和STIM1在OLP患者外周血淋巴細(xì)胞中高表達(dá),可能導(dǎo)致機體外周免疫系統(tǒng)失衡,這可能與OLP的病程遷延、反復(fù)發(fā)作有一定關(guān)系。 日的：評估脫落細(xì)胞DNA含量分析法監(jiān)測口腔黏膜潛在惡性病變癌變的靈敏度、特異度及其實用性,尋求更有效的口腔黏膜癌前診斷的無創(chuàng)性監(jiān)測手段。 方法：本實驗采用細(xì)胞刷收集了52例口腔黏膜潛在惡性病變患者的口腔黏膜病損處脫落細(xì)胞,同時切取相同部位進行病理活檢。用全自動DNA含量檢測儀檢測口腔黏膜脫落細(xì)胞的DNA含量,出現(xiàn)細(xì)胞DNA含量大于5c,診斷為陽性。以病理活檢報告作為診斷金標(biāo)準(zhǔn),評估脫落細(xì)胞DNA含量檢測的敏感度和精確度。 結(jié)果：以病理活檢報告作為診斷金標(biāo)準(zhǔn),脫落細(xì)胞DNA倍體分析的靈敏度為86.36%,特異度為90.00%,假陽性率為13.64%,假陰性率為10.00%,陽性預(yù)測價值為86.36%,陰性預(yù)測價值為90.00%。 結(jié)論：脫落細(xì)胞DNA含量檢測作為一種無創(chuàng)性監(jiān)測手段,在對口腔黏膜潛在惡性病變癌變監(jiān)測中具有較高靈敏度和特異度,患者依從性高,適于作為一種癌前監(jiān)測手段,監(jiān)測口腔黏膜潛在惡性病變病情進展?fàn)顩r。
[Abstract]:Objective: to explore the level of free calcium [Ca2] I in peripheral blood lymphocytes of oral lichen planus (OLP). And the expression levels of two closely related proteins: Golli-MBP genes of the Oligodendrocyte lineage-Myelin Basic protein (oligodendrocyte lineage gene-myelin basic protein) and STIM1(Stromal Interaction Molecule1, matrix interaction molecule 1). To investigate the effect of Golli-MBP STM1 on peripheral immune cells in oral lichen planus and to explore the pathogenesis of oral lichen planus. Methods: calcium fluorescence probe Fluo-3 was used to detect the level of free calcium ion in peripheral blood lymphocytes of 22 cases of non-erosive type and 21 cases of erosive type in 43 cases of OLP and 20 cases of normal people. The gene and protein expression of Golli-MBP and STIM1 in peripheral blood lymphocytes of 43 OLP patients and 20 normal controls were detected by quantitative PCR(Quantitive Real-Time PCR (qPCR) and immuno-protein imprinting technique. To compare whether there were statistical differences in the indexes of each group. Results: the results showed that the peripheral blood lymphocytes of OLP patients were stimulated by activation. The expression of intracellular free calcium mRNA and protein in OLP group was significantly higher than that in normal control group (P 0.001N. Golli-MBP mRNA and protein), and the expression of STIM1 mRNA and protein in OLP group was also significantly higher than that in normal control group (P 0.01). Conclusion the level of intracellular free calcium ion in peripheral blood lymphocytes of patients with OLP is lower than that of control group during activation, suggesting that the calcium signaling pathway of lymphocytes in OLP may be abnormal. This study also found that the high expression of Golli-MBP and STIM1 in peripheral blood lymphocytes of OLP patients may lead to the imbalance of peripheral immune system, which may delay the course of OLP. Repeated attacks have something to do with it. Objective: to evaluate the sensitivity, specificity and practicability of exfoliated cell DNA assay in the detection of potential malignant lesions of oral mucosa, and to seek a more effective non-invasive monitoring method for precancerous diagnosis of oral mucosa. Methods: the abscission cells were collected from 52 patients with potential malignant lesions of oral mucosa by cell brush. At the same time, the same site was removed for pathological biopsy. The DNA content of the exfoliated cells of oral mucosa was detected by automatic DNA content detector. The DNA content of the cells was more than 5c. the diagnosis was positive. The pathological biopsy report was used as the diagnostic gold standard. To evaluate the sensitivity and accuracy of DNA content detection in exfoliated cells. Results: with pathological biopsy report as diagnostic gold standard, the sensitivity, specificity, false positive rate, false negative rate, positive predictive value and negative predictive value of DNA ploidy analysis in exfoliated cells were 86.36, 90.00,13.64, 10.00, respectively, respectively. Conclusion: as a noninvasive monitoring method, the detection of DNA content in exfoliated cells has high sensitivity and specificity in the monitoring of potential malignant lesions of oral mucosa, and the patient's compliance is high, so it is suitable for precancerous monitoring. To monitor the progress of oral mucosal potential malignant lesions.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781.5

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