环状软骨前裂开对兔环状软骨生长发育的影响

目的:观察环状软骨前裂开对环状软骨生长发育的影响。方法:将幼兔环状软骨前正中裂开,8月后测量环状软骨内径,计算其管腔面积。结果:(1)与对照组比较,实验组兔环状软骨管腔面积无显著性差异(P>0.05)。(2)裂开处均由软组织连接,未形成过多瘢痕。结论:环状软骨前裂开对环状软骨及动物的生长发育无明显影响。     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

先天性眼球震颤患者眼震电图检查及评价

目的：探讨眼震电图对先天性眼震患者手术效果的评定及其临床意义。方法：对48例手术前后先天性眼震患者行眼震电图检测，中间位检查方法为令患者头正位注视正前方50cm处与双眼平行的目标光点；代偿位检查时令患者放松以平时视物最佳位为准，在此位患者眼震最小。观察中间位和代偿位眼震的速度、幅度和频率。眼震幅度、频率和速度计算均取最大眼震反应期10s时期内的平均值。结果：眼震电图可以记录到跳动型和钟摆型两种不同的眼震波，跳动型呈锯齿波，钟摆型呈正旋波。手术后中心位和代偿位的眼震与术前相比有明显降低。43例患者的眼震速度在中心位和代偿位有不同程度的下降或消失，占89．5％，48例患者中眼震幅度下降者42例，占83．3％。术后眼震频率在中心位和代偿位也均有下降，但以中心位明显。其中17例患者代偿位眼震的频率，幅度和速度降为零。术前中心位的眼震速度、幅度和频率分别为(29．64&;#177;18．22)mm／s，(8．52&;#177;3．36)mV，(9．88&;#177;1．21)Hz。术后分别下降为(12．87&;#177;9．24)mm／s(t=5．68，P&;lt;0.01)，(3．54&;#177;2．12)mV(t=8．74，P&;lt;0.01)，(2．86&;#177;1．83)Hz(t=21．93．P&;lt;0.01)。结论：眼震电图检查可以对先天性眼震患者手术效果进行客观有效的评定。     

新兴细胞生物学技术-组织微阵列研究进展

肿瘤相关基因的研究在肿瘤病理学中占有重要的地位，新兴的细胞生物技术-组织微阵列，具有批量性、快速性、准确性等特点，为肿瘤相关基因的研究提供了一种全新的方法。从组织微阵列技术的原理、主要优势、应用、研究进展、存在的问题和展望等方面作一综述。     

兔小肠黏膜下层与软骨细胞体外培养

目的观察软骨细胞在兔小肠黏膜下层(SIS)上的生长特性,为使SIS作为软骨组织工程化的载体打下基础。方法用物理和化学方法处理兔小肠黏膜下层,将不同浓度的兔软骨细胞与SIS进行体外共培养,分别进行组织学、相差显微镜和扫描电镜观察。结果经物理和化学处理的SIS纯度高、孔隙多,胶原纤维未受损;软骨细胞在SIS材料上生长、黏附、增殖,并能长入材料的孔隙内,分泌大量的细胞外基质成分。结论SIS细胞相容性良好,不影响软骨细胞的形态,对细胞生长和功能表达无抑制作用,是一种较理想的软骨细胞载体。     

异种骨载体复合骨形态发生蛋白修复兔甲状软骨缺损的实验

目的：观察应用重组合骨修复兔甲状软骨缺损的效果，探讨其作为喉气管修复材料的可行性。方法：实验于2003—10／2004—11在第四军医大学唐都医院耳鼻咽喉科教研室实验室完成。取新西兰大白兔32只，随机分成空白对照组、单纯异种骨组、经脱脂、脱钙处理的异种骨作为载体复合骨形态生成蛋白(1：20)的重组合骨组。制造兔甲状软骨缺损模型，于缺损处分别植入肌肉、单纯异种骨及重组合骨。分别于术后4，8，16周以及12个月观察局部及应用CT检查和造模处局部组织切片苏木精-伊红染色了解成骨和修复情况。结果：①局部观察：空白对照组缺损部位由瘢痕组织修复，在各时间点未见新骨形成。单纯异种骨组可见植入物被软组织包膜包裹，并逐渐被吸收，至16周基本完全吸收。重组合骨组可观察到植人物被吸收的同时有新骨诱导形成，16周局部有松质骨样组织形成，质地较硬，12个月后无明显变化。②组织学观察：见空白对照组无新骨形成，局部为新生瘢痕组织替代。单纯异种骨对照组可见局部有炎细胞浸润，植人物孔洞内有细胞长人，8周时植人物部分吸收，16周及12个月后缺损处异种骨基本完全吸收，为纤维组织替代修复，无明显新骨形成。重组合骨组4周可见植入处大量软骨组织诱导分化，在植人物松质骨空洞内可见组织细胞长人，周边有纤维组织包膜，以及炎细胞浸润，8周可见新骨形成，16周原植人物吸收，有大量骨基质形成并为纤维组织包绕。12个月后无明显变化。③CT检查：单纯异种骨与对照组8周左右大部分植入物被吸收，16周组织局部无骨组织显影。重组合骨组16周及12个月后局部有新骨形成，且气道形态维持良好，呼吸道通畅。结论：重组合骨具备支架的骨引导作用，其诱导成骨效果良好，是理想的喉气管修复材料。     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

喉移植研究进展

恶性肿瘤及严重喉损伤常导致患者喉功能丧失，给患者带来极大的生理、心理痛苦。目前的研究认为要恢复喉的功能，最理想的方法莫过于喉移植。本文就这方面研究现状及进展进行综述。     

儿童喉乳头状瘤细胞的原代培养及生物学特性研究

目的　探讨人乳头状瘤病毒 (humanpapillomavirus,HPV)感染阳性患儿喉乳头状瘤细胞在体外培养的生物学特性。方法　 2 0 0 0年 3月～ 2 0 0 1年 4月采用组织块培养法培养 10例 12例次喉乳头状瘤患儿手术的标本———HPV感染阳性患儿喉乳头状瘤细胞 ,观察其生长情况 ,计数法绘制细胞生长曲线 ,运用共同引物聚合酶链反应 (polymerasechainreaction ,PCR)法及核酸分子斑点杂交方法对喉乳头状瘤细胞在培养前后的HPVDNA进行检测。结果　HPV感染阳性的喉乳头状瘤细胞在体外生长时间可长达 6周 ,培养前后细胞内均含有HPV的DNA。细胞生长分为 3期 ,延缓期、生长期及停滞期。培养前 1～ 4d细胞从组织块中大量游出 ,5～ 7d为延缓期 ,此期间细胞渐渐开始贴壁 ,8～ 18d为生长期 ,细胞数目迅速增多 ,生长速度很快 ,继而进入停滞期 ,细胞数目增长缓慢 ,细胞空泡化明显 ,渐渐走向死亡。结论　HPV感染阳性的喉乳头状瘤细胞在体外生长情况良好 ,但要建立HPV感染阳性的喉乳头状瘤细胞的动物模型尚需进一步研究     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.