本文選題：急性呼吸窘迫綜合征(ARDS) + 肺內(nèi)源性急性呼吸窘迫綜合征(ARDSp)　； 參考：《東南大學(xué)》2017年碩士論文

【摘要】：第一部分:血漿microRNA與ARDS患者病情嚴(yán)重程度及預(yù)后相關(guān)性的研究目的:本實(shí)驗(yàn)研究血漿中MSC-VEC-miRNA(與間充質(zhì)干細(xì)胞及血管內(nèi)皮細(xì)胞相關(guān)且既往在ARDS中有過研究的miRNA)水平與ARDS患者病情嚴(yán)重程度的相關(guān)性及其對(duì)患者預(yù)后的預(yù)測(cè)價(jià)值。方法:1、MSC-VEC-miRNA篩選:利用Pubmed數(shù)據(jù)庫(kù)通過關(guān)鍵詞及主題詞檢索相關(guān)文獻(xiàn),閱讀文獻(xiàn)篩選出MSC-VEC-miRNA。2、患者入組,標(biāo)本采集:納入2016年01月至2016年09月收住東南大學(xué)附屬中大醫(yī)院重癥醫(yī)學(xué)科診斷為ARDS且符合研究納入和排除標(biāo)準(zhǔn)的患者,于患者診斷ARDS24小時(shí)內(nèi)采外周血,離心后留取血漿保存。3、患者資料收集:基本資料(年齡、身高、體重等);一般情況(診斷、APACHE Ⅱ評(píng)分、SOFA評(píng)分、ARDS病因等);氧療方式及參數(shù)(無(wú)創(chuàng)呼吸機(jī)、有創(chuàng)呼吸機(jī)及參數(shù));預(yù)后指標(biāo)(28天病死率等)。4、血漿miRNA檢測(cè):熒光定量PCR檢測(cè)血漿中MSC-VEC-miRNA的循環(huán)閾值(CT值)。5、血漿內(nèi)皮損傷標(biāo)志蛋白及炎癥因子檢測(cè):采用ELISA法檢測(cè)患者血漿中VCAM-1、vWF、TNF-α、IL-10 濃度。結(jié)果:1、篩選 MSC-VEC-miRNA,符合條件的 MSC-VEC-miRNA 有14種:miR-15a,miR-16,miR-21,miR-24,miR-26a,miR-27a,miR-27b,miR-126,miR-146a,miR-150,miR-155,miR-221,miR-223,miR-320。2、存活組與死亡組患者一般情況比較:57例患者納入研究分析,存活組39例,死亡組18例,28天病死率為31.6%。存活組與死亡組間比較:年齡、BMI、基礎(chǔ)疾病、ARDS病因、呼氣末正壓(PEEP)、P02/Fi02無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。死亡組患者APACHE Ⅱ評(píng)分(26.9±8.5)、SOFA評(píng)分(13.7±3.9)和血乳酸(3.1[1.2-5.2])顯著高于存活組(18.7±7.1,P0.001)、(8.9±4.5,P0.001)、(1.7[0.9-2.4],P=0.01)。而死亡組患者M(jìn)urray評(píng)分顯著高于存活組(2.8[2.3-3.7]vs.2.3[1.7-2.7],P=0.01)。3、存活組與死亡組患者血漿MSC-VEC-miRNA、血管內(nèi)皮損傷蛋白及炎癥因子比較:ARDS死亡組患者血漿miR-26a水平明顯低于存活組(0.33[0.09-1.17]vs.0.97[0.17-3.49],P=0.046),而 ARDS 死亡組患者血漿 miR-320水平明顯高于存活組(0.37[0.16-1.66]vs.0.18[0.07-0.39],P=0.041),其他 12 種miRNA在存活組與死亡組血漿中水平無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。存活組與死亡組血漿 vWF、VCAM-1、TNF-α、IL-10 濃度無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。4、不同ARDS嚴(yán)重程度患者血漿MSC-VEC-miRNA、血管內(nèi)皮損傷蛋白及炎癥因子比較:納入患者中,ARDS輕度13例,中度25例,重度19例,輕、中、重度ARDS三組患者血漿MSC-VEC-miRNA水平無(wú)統(tǒng)計(jì)學(xué)差異(P0.05);三組患者血漿vWF、VCAM-1、TNF-α、IL-10水平無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。5、血漿MSC-miRNA對(duì)ARDS28天預(yù)后的預(yù)測(cè)價(jià)值:(1)miR-26a:AUC=0.67,p0.05;(2)miR-320:AUC=0.67,p0.05;(3)聯(lián)合 miR-26a、miR-320:AUC=0.75,p0.001。結(jié)論:血漿MSC-VEC-miRNA水平與ARDS嚴(yán)重程度無(wú)相關(guān)性,血漿miR-26a、miR-320對(duì)ARDS患者28天預(yù)后有一定的預(yù)測(cè)價(jià)值。第二部分:肺內(nèi)源性ARDS和肺外源性ARDS患者血漿microRNA表達(dá)水平差異的研究目的:本實(shí)驗(yàn)探究MSC-VEC-miRNA(與間充質(zhì)干細(xì)胞及血管內(nèi)皮細(xì)胞相關(guān)且既往在ARDS中有過研究的miRNA)在肺內(nèi)源性ARDS和肺外源性ARDS患者血漿中水平差異。方法:1、MSC-VEC-miRNA篩選:利用Pubmed數(shù)據(jù)庫(kù)通過關(guān)鍵詞及主題詞檢索相關(guān)文獻(xiàn),通過閱讀文獻(xiàn)篩選出MSC-VEC-miRNA。2、患者入組,標(biāo)本采集:納入2016年01月至2016年09月收住東南大學(xué)附屬中大醫(yī)院重癥醫(yī)學(xué)科診斷為ARDS患者并符合本研究納入和排除標(biāo)準(zhǔn)的患者57例,于患者診斷ARDS 24小時(shí)內(nèi)采外周血,離心后留取血漿并保存。3、患者資料收集:基本資料(年齡、身高、體重等);一般情況(診斷、APACHE Ⅱ評(píng)分、SOFA評(píng)分、ARDS病因等);氧療方式及參數(shù)(無(wú)創(chuàng)呼吸機(jī)、有創(chuàng)呼吸機(jī)及參數(shù));預(yù)后指標(biāo)(28天病死率等)。4、血漿miRNA檢測(cè):熒光定量PCR檢測(cè)出血漿中MSC-VEC-miRNA的循環(huán)閾值(CT值)。5、血漿內(nèi)皮損傷標(biāo)志蛋白及炎癥因子檢測(cè):采用ELISA法檢測(cè)患者血漿中VCAM-1、vWF、TNF-α、IL-10濃度。結(jié)果:1、篩選 MSC-VEC-miRNA,符合條件 MSC-VEC-miRNA有14種:miR-15a,miR-16,miR-21,miR-24,miR-26a,miR-27a,miR-27b,miR-126,miR-146a,miR-150,miR-155,miR-221,miR-223,miR-320。2、肺內(nèi)與肺外源性ARDS患者臨床病情指標(biāo)比較:符合標(biāo)準(zhǔn)的57例ARDS患者納入研究分析,其中肺內(nèi)源性ARDS 43例,肺外源性ARDS 14例。肺內(nèi)源性ARDS患者與肺外源性ARDS患者一般情況比較:兩組間年齡、性別、BMI、基礎(chǔ)疾病、28天病死率、APACHE Ⅱ評(píng)分、SOFA評(píng)分、乳酸無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。肺內(nèi)源性ARDS患者與肺外源性ARDS患者ARDS嚴(yán)重程度比較:PO2/FiO2在肺內(nèi)源性ARDS組明顯低于肺外源性ARDS組(145[119-203]vs.206[184-253],P=0.012);Murray肺損傷評(píng)分在肺內(nèi)源性ARDS組顯著高于肺外源性 ARDS 組(2.7[2-3.3]vs.1.8[1.3-2.4],P=0.008),兩組間 FiO2、PEEP無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。3、肺內(nèi)與肺外源性ARDS患者血漿MSC-VEC-miRNA比較:肺外源性ARDS 患者血漿肺外源性 ARDS 患者血漿 miR-221(0.22[0.12-0.49])、miR-27b(0.34[0.10-0.46])水平顯著低于肺內(nèi)源性 ARDS 患者(0.57[0.22-1.57],P=0.008)、(0.60[0.20-1.46],P=0.025),其他miRNA在兩組患者血漿中表達(dá)水平無(wú)明顯差異(P0.05)。4、肺內(nèi)與肺外源性ARDS患者血管內(nèi)皮損傷蛋白及炎癥因子比較:肺外源性ARDS組患者血漿vWF顯著低于肺內(nèi)源性ARDS患者(0.77[0.29-1.54]vs.1.80[0.95-3.51],P=0.048);VCAM-1、IL-10、TNF-α 在兩組患者血漿中的濃度無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論:miR-221、miR-27b、vWF在肺外源性ARDS患者血漿水平顯著低于肺內(nèi)源性ARDS患者。
[Abstract]:Part one: Study of the correlation between the severity and prognosis of patients with plasma microRNA and ARDS. Objective: To study the correlation between the level of plasma MSC-VEC-miRNA (related to mesenchymal stem cells and vascular endothelial cells and the previously studied miRNA in ARDS) and the severity of the severity of the patients with ARDS and the prognosis of the patients Methods: 1, MSC-VEC-miRNA screening: using the Pubmed database to retrieve the relevant literature through the key words and subject words, and read the literature to screen out MSC-VEC-miRNA.2, the patients into the group, the specimen collection: from 01 months to 09 months of 2016, the medical department of Zhongda Hospital Affiliated to Southeast University in the severe medical department was diagnosed as ARDS and accords with the inclusion and exclusion criteria. Patients received peripheral blood in ARDS24 hours and retained plasma after centrifugation and retained.3 after centrifugation. The data were collected: basic data (age, height, weight, etc.); general conditions (diagnosis, APACHE II score, SOFA score, ARDS etiology, etc.); oxygen therapy and parameters (non-invasive ventilator, invasive ventilator and parameters); prognosis index (28 days fatality rate, etc.). 4, plasma miRNA detection: fluorescence quantitative PCR detection of plasma MSC-VEC-miRNA cycle threshold (CT value).5, plasma endothelial damage marker protein and inflammatory factors detection: ELISA method for detecting VCAM-1, vWF, TNF- alpha, IL-10 concentration in patients plasma. Results: 1, screening MSC-VEC-miRNA, 14 types of MSC-VEC-miRNA. R-24, miR-26a, miR-27a, miR-27b, miR-126, miR-146a, miR-150, miR-155, miR-221, miR-223, miR-320.2, the general comparison between the survival group and the death group: 57 cases were included in the study analysis, the survival group 39 cases, the death group 18 cases, the 28 day fatality rate were compared between the 31.6%. survival group and the death group: age, basic disease, etiology, end expiratory pressure ( PEEP), there was no statistical difference in P02/Fi02 (P0.05). The score of APACHE II in the death group was (26.9 + 8.5), the SOFA score (13.7 + 3.9) and blood lactic acid (3.1[1.2-5.2]) were significantly higher than that in the survival group (18.7 + 7.1, P0.001), (8.9 + 4.5, P0.001), and (1.7[0.9-2.4], P=0.01). Compared with the plasma MSC-VEC-miRNA, vascular endothelial damage protein and inflammatory factors in the survival group and the death group, the plasma miR-26a level of the patients in the ARDS death group was significantly lower than that in the survival group (0.33[0.09-1.17]vs.0.97[0.17-3.49], P=0.046), while the plasma miR-320 level in the ARDS death group was significantly higher than that in the survival group (0.37[0.16-1.66]vs.0.18[0.07-0.39], P=). 0.041) there was no significant difference in the plasma levels of the other 12 kinds of miRNA in the survival group and the death group (P0.05). The plasma vWF, VCAM-1, TNF-, and IL-10 concentrations in the survival group and the death group were not statistically different (P0.05).4, and the plasma MSC-VEC-miRNA, vascular endothelial damage protein and inflammatory factors in patients with different ARDS severity were compared: ARDS mild 13 was included in the patients. There were 25 cases of moderate, 19 severe, moderate, moderate, and severe ARDS three groups with no statistically significant difference in plasma MSC-VEC-miRNA (P0.05); the plasma vWF, VCAM-1, TNF- a, IL-10 levels were not statistically different (P0.05) in the three groups, and the predictive value of plasma MSC-miRNA to ARDS28 day prognosis: (1) miR-26a:AUC=0.67, 2); (3) union 26a, miR-320:AUC=0.75, p0.001. conclusion: there is no correlation between plasma MSC-VEC-miRNA level and ARDS severity. Plasma miR-26a and miR-320 have a certain predictive value for the 28 day prognosis of ARDS patients. The second part: Study on the difference of plasma microRNA expression level in pulmonary endogenous ARDS and pulmonary ARDS patients: this experiment explored MSC-VEC-miRNA (and between) The levels of plasma levels of miRNA in the endogenous ARDS and pulmonary exogenous ARDS patients in ARDS were related to endothelium derived stem cells and vascular endothelial cells. Methods: 1, MSC-VEC-miRNA screening: using the Pubmed database to retrieve the relevant literature through the key words and subject words, and to select the MSC-VEC-miRNA.2 and the patients into the group through the reading literature. Sample collection: 57 patients, diagnosed as ARDS patients in the Department of severe medicine of Zhongda Hospital Affiliated to Southeast University from 01 to 2016 2016, were diagnosed as patients and were eligible for the inclusion and exclusion of this study. The peripheral blood was collected within 24 hours of the diagnosis of ARDS, the plasma was retained after centrifugation and.3 was preserved, and the data were collected: basic data (age, height, body). Heavy wait); general conditions (diagnosis, APACHE II score, SOFA score, ARDS etiology, etc.); oxygen therapy and parameters (non-invasive ventilator, invasive ventilator and parameters); prognostic index (28 days of mortality, etc.).4, plasma miRNA detection: fluorescence quantitative PCR detection of MSC-VEC-miRNA cycle threshold (CT).5, plasma endothelial damage marker protein and inflammation ELISA method was used to detect VCAM-1, vWF, TNF- alpha and IL-10 concentration in patients' plasma. Results: 1, screening MSC-VEC-miRNA, miR-15a, miR-16, miR-21, miR-24, miR-26a. Index comparison: 57 cases of ARDS patients who met the standard were included in the study, including 43 pulmonary endogenous ARDS cases and 14 pulmonary ARDS cases. Pulmonary endogenous ARDS patients and pulmonary exogenous ARDS patients were compared: the two groups were age, sex, BMI, basic disease, 28 day mortality, APACHE II score, SOFA score, and no statistical difference in lactic acid (P0.05). Lung The severity of ARDS in endogenous ARDS patients and pulmonary exogenous ARDS patients was compared: PO2/FiO2 in endogenous ARDS group was significantly lower than that of pulmonary ARDS group (145[119-203]vs.206[184-253], P=0.012), and Murray lung injury score in the pulmonary endogenous ARDS group was significantly higher than that in the pulmonary ARDS group (2.7[2-3.3]vs.1.8[1.3-2.4], P=0.012). No statistical difference (P0.05).3, plasma MSC-VEC-miRNA in patients with pulmonary and pulmonary exogenous ARDS: plasma miR-221 (0.22[0.12-0.49]) and miR-27b (0.34[0.10-0.46]) in pulmonary exogenous ARDS patients were significantly lower than that of lung endogenous ARDS (0.57 [0.22-1.57]). There was no significant difference in plasma expression level in the two groups (P0.05).4. The vascular endothelial damage protein and inflammatory factors in the pulmonary and exogenous ARDS patients were compared: the plasma vWF of the pulmonary exogenous ARDS patients was significantly lower than that of the pulmonary endogenous ARDS patients (0.77[0.29-1.54]vs.1.80[0.95-3.51], P= 0.048); VCAM-1, IL-10, TNF- alpha was in the plasma of two groups of patients. There was no significant difference in concentration (P0.05). Conclusion: miR-221, miR-27b and vWF in plasma of patients with extrinsic ARDS are significantly lower than those in patients with endogenous ARDS.

【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563.8

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