本文選題：髓核細(xì)胞 + 細(xì)胞鑒定　； 參考：《昆明醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的：CD24是一種低分子質(zhì)量高度糖基化的粘附分子,表達(dá)于人體多種細(xì)胞表面。該研究我們通過對比人髓核與軟骨中CD24的表達(dá),探討CD24在鑒別髓核細(xì)胞中的作用及效果。方法：通過收集手術(shù)治療中患者的椎間盤及軟骨組織,患者年齡5～71歲,共27例,手術(shù)原因包括先天性脊柱側(cè)凸、外傷、椎間盤退變。其中標(biāo)本組織切片HE染色觀察組織,另一部分標(biāo)本的髓核組織和軟骨組織提取RNA后逆轉(zhuǎn)錄為cDNA進(jìn)行OD值測定,普通凝膠電泳PCR、RT-PCR測定組織中CD24的表達(dá)情況并進(jìn)行對比。結(jié)果：1.大體觀察：健康髓核外觀呈乳白或灰白色半透明或不透明的膠凍樣物質(zhì),富有彈性.退變髓核組織分界清晰,髓核經(jīng)髓核鉗咬出后呈絮狀灰白或淡黃色組織,其組織彈性稍差2.組織切片HE染色：髓核組織結(jié)構(gòu)清晰,纖維環(huán)染色均勻,髓核細(xì)胞呈巢狀排列,無退變患者髓核細(xì)胞數(shù)量豐富,細(xì)胞形態(tài)規(guī)整,細(xì)胞突長度適中,輕度退變髓核組織內(nèi)細(xì)胞數(shù)量較正常髓核內(nèi)輕度減少,細(xì)胞形態(tài)尚規(guī)整,退變組織纖維環(huán)內(nèi)偶見少量巨噬細(xì)胞。軟骨組織染色均勻,可見軟骨陷凹,軟骨細(xì)胞染色較深未見其他組織結(jié)構(gòu)。3.RNA紫外分光度儀器檢測結(jié)果：本次實(shí)驗(yàn)中所提取的RNA經(jīng)檢測OD值與濃度,提示RNA純度較高,濃度滿足后續(xù)實(shí)驗(yàn)要求。4.普通凝膠電泳熒光PCR結(jié)果：可見髓核與軟骨內(nèi)參基因產(chǎn)物條帶位置大約為200dp左右,條帶清晰,見髓核CD24通過PCR擴(kuò)增后產(chǎn)物有清晰條帶位置大概為150dp,而軟骨CD24特異性擴(kuò)增處未見明顯條帶。5.RT-PCR結(jié)果：實(shí)驗(yàn)各組髓核內(nèi)CD24表達(dá)與軟骨內(nèi)表達(dá)差異均有顯著統(tǒng)計學(xué)意義(PO.01),各組髓核內(nèi)CD24的表達(dá)差異沒有統(tǒng)計學(xué)意義(PO.05),髓核內(nèi)CD24的表達(dá)與年齡之間沒有明顯相關(guān)關(guān)系(相關(guān)系數(shù)rs=0.124,P0.05)結(jié)論：1. Pfirrmann Ⅰ～Ⅲ級髓核組織主要細(xì)胞成分為髓核細(xì)胞；2.CD24在髓核組織與軟骨組織之間表達(dá)存在明確的統(tǒng)計學(xué)差異,CD24可以作為Ⅱ型膠原蛋白和蛋白聚糖之后的特異性鑒定因子；3.不同年齡、不同疾病類型、Pfirrmann Ⅰ～Ⅲ型的髓核組織內(nèi)CD24的表達(dá)沒有統(tǒng)計學(xué)意義,CD24與髓核細(xì)胞的功能及退變程度沒有明顯相關(guān)性；
[Abstract]:Objective: to express the low molecular weight and high glycosylated adhesion molecule on the surface of many human cells. In this study, we compared the expression of CD24 in nucleus pulposus and cartilage to explore the role and effect of CD24 in differentiating nucleus pulposus cells. Methods: 27 patients (571 years old) with congenital scoliosis, trauma and disc degeneration were collected. The tissue sections were stained with HE, and the other part of the tissue of nucleus pulposus and cartilage were extracted from the nucleus pulposus and chondrocytes. The OD value was measured by reverse transcription to cDNA. The expression of CD24 in the tissue was detected by RT-PCR and compared. The result is 1: 1. General observation: the appearance of healthy nucleus pulposus is milky white or grayish-white translucent or opaque gelatinous material, elastic. The tissue boundary of degenerative nucleus pulposus was clear. The nucleus pulposus showed flocculent grayish white or light yellow tissue after bite of nucleus pulposus forceps, and its tissue elasticity was slightly less than 2. HE staining showed clear structure of nucleus pulposus, uniform staining of fibrous ring, nesting arrangement of nucleus pulposus cells, abundant number of nucleus pulposus cells, regular morphology of cells and moderate length of cell processes in patients without degeneration. The number of cells in the mild degenerative nucleus pulposus was slightly lower than that in the normal nucleus pulposus, and the morphology of the cells was still regular, and a few macrophages were occasionally found in the fibrous ring of the degenerative tissue. Cartilage tissue staining was uniform, cartilage concave was visible, and other tissue structures were not found in chondrocyte staining. 3. The RNA extracted in this experiment was detected by OD value and concentration, indicating that the purity of RNA was high. The concentration meets the requirements of subsequent experiments. The results of fluorescence PCR showed that the position of gene products in nucleus pulposus and cartilage was about 200dp and the bands were clear. It was found that the clear band position of CD24 amplified by PCR was about 150dp, but there were no obvious bands in the specific amplification of cartilage CD24. 5. RT-PCR results: there were significant differences between the expression of CD24 in nucleus pulposus and the expression in cartilage in each group. 5. The results of RT-PCR showed that there were significant differences in the expression of CD24 in the nucleus pulposus and in the cartilage. There was no significant difference in the expression of CD24 in nucleus pulposus (P < 0.05). There was no significant correlation between the expression of CD24 in nucleus pulposus and age (r = 0.124, P 0.05). Conclusion: 1. The main cell component of Pfirrmann 鈪，

本文編號：1924208