本文選題：先天性脊柱側(cè)彎 + 頂椎�。� 參考：《鄭州大學(xué)》2012年碩士論文

【摘要】：先天性脊柱側(cè)彎(Congenital scoliosis,CS)是臨床上較常見(jiàn)的一種小兒先天性脊柱畸形,其發(fā)生發(fā)展主要?dú)w因于先天存在的異常脊椎導(dǎo)致脊柱縱向不平衡生長(zhǎng)。在脊柱縱向發(fā)育過(guò)程中脊椎生長(zhǎng)板軟骨是縱向生長(zhǎng)的基礎(chǔ),頂椎作為先天性脊柱側(cè)彎的觸發(fā)點(diǎn),其生長(zhǎng)板軟骨的化骨活性直接關(guān)系到側(cè)彎的發(fā)生發(fā)展情況。以往研究已證實(shí)脊椎生長(zhǎng)板具有類(lèi)似長(zhǎng)骨生長(zhǎng)板的特性,生長(zhǎng)分化受多種細(xì)胞因子的調(diào)控。其中SOX9是一個(gè)與哺乳動(dòng)物的性別決定因子SRY(Y染色體上的性別決定區(qū))具有高度同源性的轉(zhuǎn)錄因子,表達(dá)于除肥大軟骨細(xì)胞外所有分化型軟骨細(xì)胞,并貫穿整個(gè)軟骨形成過(guò)程,被認(rèn)為在軟骨內(nèi)成骨的初始階段發(fā)揮著重要作用。血管內(nèi)皮生長(zhǎng)因子(VEGF)是刺激內(nèi)皮細(xì)胞生長(zhǎng)和血管再生的一個(gè)重要因素,研究發(fā)現(xiàn)在軟骨內(nèi)化骨過(guò)程中對(duì)于軟骨細(xì)胞死亡,細(xì)胞外基質(zhì)重塑,血管生成和骨骼形成是一種基本的調(diào)節(jié)物質(zhì)。因此通過(guò)對(duì)側(cè)彎頂椎凸凹兩側(cè)生長(zhǎng)板軟骨VEGF和SOX9表達(dá)情況的檢測(cè)可以判斷側(cè)彎兩側(cè)軟骨板成骨能力的差異,推斷小兒先天性脊柱側(cè)彎發(fā)病和進(jìn)展的分子生物學(xué)基礎(chǔ),為進(jìn)一步的分子生物學(xué)研究奠定基礎(chǔ)。 目的 通過(guò)免疫組化和：mRNA原位雜交方法判定先天性脊柱側(cè)彎患兒凸凹兩側(cè)生長(zhǎng)板軟骨SOX9和VEGF的表達(dá)部位和情況,了解兩側(cè)生長(zhǎng)板軟骨生物活性的差異,發(fā)現(xiàn)SOX9和VEGF表達(dá)的相關(guān)性,探討影響先天性脊柱側(cè)彎發(fā)病和進(jìn)展的分子生物學(xué)基礎(chǔ)。 方法 取2010年10月～2011年9月我院收住的20例1～5歲先天性脊柱側(cè)凸患者,男8例,女12例,平均年齡3.2(3.2±0.9)歲,術(shù)前訂均接受詳細(xì)的體格檢查和影像學(xué)檢查(包括x線片,全脊柱螺旋CT+三維重建,MRI)。入院后經(jīng)患者直系親屬的知情同意后,取術(shù)中切除的半椎體凸凹兩側(cè)生長(zhǎng)板軟骨,常規(guī)HE染色并應(yīng)用免疫組化和mRNA原位雜交的方法對(duì)頂椎生長(zhǎng)板的SOX9和VEGF表達(dá)進(jìn)行檢測(cè),觀察SOX9和VEGF在頂椎生長(zhǎng)板軟骨的表達(dá)位置,以及在頂椎凸凹兩側(cè)生長(zhǎng)板軟骨的表達(dá)情況,同時(shí)對(duì)SOX9、VEGF和VEGFmRNA的表達(dá)進(jìn)行相關(guān)性研究。凸凹兩側(cè)因子表達(dá)結(jié)果采用配對(duì)t檢驗(yàn),p0.05為差異有統(tǒng)計(jì)學(xué)意義。相關(guān)性分析采用Pearson相關(guān)性分析,以p0.05為檢驗(yàn)水準(zhǔn)。 結(jié)果 l、凸凹兩側(cè)生長(zhǎng)板的形態(tài)學(xué)觀察,兩側(cè)軟骨板HE染色可見(jiàn)生長(zhǎng)板有類(lèi)似長(zhǎng)骨生長(zhǎng)板的結(jié)構(gòu),大致可以分為靜止層、增殖層和肥大層軟骨區(qū)。凸側(cè)生長(zhǎng)板軟骨的增殖區(qū)和肥大區(qū)厚度更大,細(xì)胞排列也較緊密。 2、SOX9主要表達(dá)于靜止層和增殖層軟骨細(xì)胞,VEGF主要表達(dá)肥大軟骨區(qū),少量表達(dá)于增殖層,但靜止層未見(jiàn)表達(dá)。SOX9在凸側(cè)生長(zhǎng)板軟骨表達(dá)量為154.376±8.055,凹側(cè)為121.413±6.751,VEGF在凸凹兩側(cè)生長(zhǎng)板軟骨的表達(dá)量分別為134.989±9.731,108.498±8.737,VEGFmRNA在凸凹兩側(cè)生長(zhǎng)板的表達(dá)為116.517±7.536和97.077±6.627,分別經(jīng)配對(duì)t檢驗(yàn)p均0.05, SOX9、VEGF、 VEGFmRNA在頂椎凸凹兩側(cè)生長(zhǎng)板軟骨的表達(dá)差異有統(tǒng)計(jì)學(xué)意義。 3.對(duì)脊椎生長(zhǎng)板軟骨SOX9和VEGF的表達(dá)進(jìn)行Pearson相關(guān)性分析,r=0.650,p0.05,有統(tǒng)計(jì)學(xué)意義。對(duì)VEGF的表達(dá)和VEGFmRNA的表達(dá)進(jìn)行相關(guān)性分析,r=0.343,p0.05。無(wú)統(tǒng)計(jì)計(jì)學(xué)意義。 結(jié)論 1、先天性脊柱側(cè)彎患者頂椎生長(zhǎng)板軟骨,大體觀凸側(cè)較凹側(cè)具有較高的生物活性。 2、SOX9、VEGF和VEGFmRNA在先天性脊柱側(cè)彎患兒頂椎凸側(cè)生長(zhǎng)板軟骨的表達(dá)量較凹側(cè)高,這種表達(dá)差異可能是小兒先天性脊柱側(cè)彎發(fā)病和進(jìn)展的關(guān)鍵因素。 3、SOX9和VEGF在脊椎生長(zhǎng)板不同軟骨區(qū)表達(dá)量不同。整體分析,SoX9和VEGF的表達(dá)存在正線性相關(guān)。
[Abstract]:Congenital spinal scoliosis (Congenital scoliosis, CS) is a common congenital spinal deformity in children. Its development is mainly attributable to the congenital abnormal spine resulting in the longitudinal uneven growth of the spine. In the longitudinal development of the spine, the vertebral growth plate cartilage is the basis of longitudinal growth and the apex vertebrae as a congenital spinal column. The triggering point of the lateral bending, the osteogenic activity of the growth plate cartilage is directly related to the occurrence and development of the lateral bending. Previous studies have confirmed that the vertebral growth plate has the characteristics similar to the long bone growth plate, and the growth and differentiation is regulated by a variety of cytokines. SOX9 is a sex determinant on the Y chromosome (sex determination on the Y chromosome). A highly homologous transcription factor, expressed in all differentiated chondrocytes except the hypertrophic chondrocytes and throughout the process of cartilage formation, is considered to play an important role in the initial stage of endochondral osteogenesis. Vascular endothelial growth factor (VEGF) is an important factor in stimulating the growth of the inner skin cells and the regeneration of blood vessels. In the process of cartilaginous endochondral bone, the death of cartilage cells, the remodeling of extracellular matrix, angiogenesis and bone formation is a basic regulating substance in the process of endochondral osseous bone. Therefore, the difference in the osteogenesis ability of the cartilaginous plates on both sides of the lateral bending can be judged by the detection of the expression of VEGF and SOX9 on the sides of the growth plate cartilage on both sides of the lateral bending top vertebra. Molecular biological basis for pathogenesis and progression of scoliosis in nature will lay a foundation for further molecular biology research.
objective
The expression of SOX9 and VEGF in the growth plate cartilage on both sides of the children with congenital scoliosis was determined by immunohistochemistry and mRNA in situ hybridization. The differences in the biological activity of the cartilage on both sides of the growth plate were understood, the correlation between the expression of SOX9 and VEGF was found, and the molecular biological basis of the pathogenesis and progression of congenital scoliosis was discussed.
Method
From October 2010 to September 2011, 20 cases of 1~5 year old congenital scoliosis were collected from our hospital, 8 men and 12 women, with an average age of 3.2 (3.2 + 0.9) years old. A detailed physical examination and imaging examination (including X-ray, total spinal spiral CT+ 3D reconstruction, MRI) were accepted before the operation. The expression of SOX9 and VEGF in the apical growth plate was detected by routine HE staining and immunohistochemistry and mRNA in situ hybridization. The expression of SOX9 and VEGF at the apical growth plate cartilage, and the expression of the long plate cartilage on both sides of the apical convex concave were observed, and SOX9, VEGF and VEG were also used for SOX9, VEGF and VEG on both sides of the excised hemivertebra. The expression of FmRNA was studied. The results of both sides of convex and concave factors were detected by paired t, and the difference between P0.05 was statistically significant. The correlation analysis adopted Pearson correlation analysis and P0.05 as the test level.
Result
L, the morphological observation of the growth plates on both sides of the convex concave. The structure of the growth plate similar to the long bone growth plate was observed on both sides of the cartilage plate, which could be divided into the static layer, the proliferating layer and the hypertrophic region. The proliferation area and the hypertrophic area of the protruded growth plate cartilage were more thick and the cells were arranged closely.
2, SOX9 is mainly expressed in the static and proliferating layer cartilage cells. VEGF mainly expresses the hypertrophic cartilage region, which is expressed in a small amount in the proliferation layer, but the expression of.SOX9 in the protruded growth plate cartilage is not 154.376 + 8.055, the concave side is 121.413 + 6.751, and the expression of VEGF on both sides of the convex concave is 134.989 + 9.731108.498 + 8.73, respectively. 7, the expression of VEGFmRNA on both sides of the convex concave is 116.517 + 7.536 and 97.077 + 6.627. The expression of P 0.05, SOX9, VEGF and VEGFmRNA on both sides of the top vertebral convex concave is statistically significant.
3. the expression of SOX9 and VEGF in the spinal growth plate cartilage was analyzed by Pearson correlation, r=0.650, P0.05, and there were statistical significance. The correlation analysis between the expression of VEGF and the expression of VEGFmRNA, r=0.343, p0.05. had no statistical significance.
conclusion
1, the growth plate cartilage of the apical vertebrae in congenital scoliosis has a higher biological activity than the concave side.
2, SOX9, VEGF and VEGFmRNA in children with congenital scoliosis are more expressed in the protruding plate cartilage than the concave side. This difference may be the key factor for the onset and progression of children's congenital scoliosis.
3, the expression levels of SOX9 and VEGF in different cartilage regions of the spinal growth plate were different. Overall analysis showed that there was a positive linear correlation between SoX9 and VEGF expression.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R726.5

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