口腔颌面部鳞癌尿激酶及其抑制剂的基因表达和临床意义

目的:探讨uPA、PAI-1、PAI-2mRNA表达水平与口腔颌面部鳞癌浸润、转移的关系.方法:RT-PCR法检测30例口腔颌面部鳞癌uPA、PAI-1、PAI-2mRNA表达水平.结果:癌组织uPA、PAI-1、PAI-2的mRNA含量高于相应正常组织(P＜0.01),uPA和PAI-1 mRNA的表达呈正相关(r=0.53,P＜0.05).伴有淋巴结转移癌组织uPA、PAI-1高于无淋巴结转移者(P＜0.05),uPA、PAI-1、PAI-2表达水平与T分期和分化程度无明显关系.结论:uPA、PAI-1、PAI-2表达显著升高是口腔颌面部鳞癌的重要特征,其表达水平可作为判断肿瘤性质、转移能力的参考指标.     

口腔鳞癌组织中尿激酶型纤溶酶原激活剂及其抑制剂活性与颈淋巴结转移的关系

Chromogenic substrate assay was used to determine the activity of uPA and PAI-1 in tumor extract, their relationship with clinlcal and pathological parameters were analysed. It was found that activity of uPA and PAI-1 in tumor tissue was much higher than those in the corresponding normal tissue(P＜0.01); There was no relationship between activity of uPA and PAI-1 and size, histalogical classification of tumor, but the activity of uPA had positive relation to clinical stage; The uPA and PAI-1 activity in tumor with metastasis were significantly higher than those in nonmetastasis tumor(P＞0.01),furthermore, it was shown that uPA and PAI-1 in metastastic lymphnode were greatly higher than that in primary tumor. The result indicated elevation of activity of uPA and PAI-1 might be responsible for the metastasis to neck lymphnode.     

尿激酶型纤溶酶原激活剂及其抑制剂在舌鳞癌中的表达及意义

目的:研究尿激酶型纤溶酶原激活剂(uPA)及其抑制剂(PAI-1)在舌鳞癌(TSCC)中的表达及其与临床病理参数的关系.方法:应用链霉卵白素-生物素复合体方法检测uPA及PAI-1在74例TSCC组织和15例癌旁正常黏膜中的表达.以SPSS10.0软件包对数据进行统计学处理,Kappa一致性法分析uPA和PAI-1表达...     

颞下颌关节盘前移位后关节盘组织变化的实验研究

目的:观察小型猪颞下颌关节盘前移位后关节盘的组织学变化。方法:实验用小型猪10只,2只为健康对照组,术前处死。实验组8只,左侧为实验侧,右侧为实验对照侧,在左侧用正畸橡皮筋以0.78 N经眶持续牵引关节盘向前,造成关节盘前移位动物模型。术后4、6、8、10周各处死实验动物。对关节盘标本进行组织学观察。结果:HE染色见对照组关节盘表面光滑。实验组关节盘组织出现裂纹,结构模糊不清。甲苯胺兰染色见对照组表面光滑规整,关节盘内细胞排列规则紧。实验组关节盘软骨细胞增多和灶性聚集,纤维排列紊乱。SP法检测PCNA见对照组关节盘的表面滑膜组织及滑膜下层成纤维细胞无PCNA表达。实验组关节盘内可见部分细胞呈圆形或椭圆形,其内PCNA阳性。SP法检测S-100见对照组关节盘内软骨细胞无S-100表达。实验组可见下板部分成纤维细胞及纤维软骨细胞S-100弱表达。结论:猪关节盘前移位动物模型可较好模拟人的不可复性关节盘前移位关节盘的组织病理及组化变化,组织病理学变化呈渐进性。关节盘修复是一个适应性改建过程。     

颞下颌关节盘前移位后关节组织中S-100的表达

目的：探讨颞下颌关节盘前移位后关节组织中S-100表达的变化及其意义。方法：26只日本大耳白兔，在建立颞下颌关节盘前移位动物模型后，分别于术后1周、2周、4周、6周、8周、10周和12周处死，用免疫组织化学方法检测关节组织内S-100的分布。结果：正常时S-100的表达主要位于关节盘前带和后带的软骨细胞中，双板区内无软骨细胞亦无S-100的表达。1周时下板内有少量成纤维细胞和纤维软骨细胞弱表达S-100，2周时出现少量弱表达S-100的游离软骨细胞。以后软骨细胞数目逐渐增多、S-100表达逐渐增强。10周时，可见多数强表达S-100的软骨细胞，12周时，滑膜层亦可见软骨细胞的出现及S-100的表达。结论：关节盘前移位后双板区出现软骨细胞及S-100的表达，S-100的表达可能与双板区组织的适应性改建有关。     

人口腔鳞癌裸鼠移植瘤中u-PA系统的表达及意义

目的 研究u-PA系统三种主要成分（u-PA、PAI-1及PAI-2）在口腔鳞癌细胞裸鼠移植瘤侵袭转移过程中的表达及作用。方法 将2株口腔鳞癌细胞GNM细胞和TSCCa细胞接种入裸鼠体内，建立体内转移模型。应用免疫组化法，流式细胞仪检测移植瘤组织中u-PA系统各因子的表达。结果 GNM细胞所形成移植瘤的侵袭转移能力最强；免疫组化和流式细胞仪检测均表明u-PA系统各因子在两株移植瘤肿瘤的胞浆和胞膜均有相当的表达，在GNM细胞所形成移植瘤中u-PA呈高水平表达或弱阳性表达。结论 u-PA的异常表达可能是反映两株细胞移植瘤裸鼠体内侵袭转移能力差异的指标之一。     

LPS调控人牙龈成纤维细胞表达uPA的研究

目的：观察LPS对牙龈成纤维细胞表达尿激酶型纤溶酶原激活剂（uPA)的影响。方法：采用Western blotting和Northern blotting分别观察LPS对牙龈成纤维细胞内uPA蛋白质表达水平和mRNA表达水平的影响。结果：经1.0μg/mL LPS处理8h后，牙龈成纤维细胞内uPA蛋白表达量明显增强，且这一增强作用可持续24h；经1.0μg/mL LPS处理4h后，牙龈成纤维细胞内uPA mRNA表达量明显增强。结论：LPS能够促进牙龈成纤维细胞表达uPA，参与牙周组织的破坏。     

口腔黏膜下纤维性变中u-PA和PAI-1的表达及意义

目的:了解尿激酶型纤溶酶原激活物(u-PA)及其抑制剂(PAI-1)在口腔黏膜下纤维性变(OSF)组织中的表达及其意义。方法:采用免疫组化染色法和图像分析法,对OSF早、中、晚期各10例及10例正常口腔黏膜组织中的u-PA及PAI-1的表达及分布进行分析。结果:OSF病变组织中u-PA和PAI-1有异常表达,其阳性物质主要分布于固有层血管内皮细胞、成纤维细胞及炎症细胞的胞质,PAI-1各期阳性表达强度均高于正常口腔黏膜组(P<0.05),u-PA早期阳性表达显著升高(P<0.05),中晚期则逐渐降低(与正常组比P>0.05)。结论:u-PA、PAI-1在OSF病变组织中的异常表达,可能是影响OSF发生发展的重要因素。     

口腔鳞癌中u-PA系统的表达与颈淋巴结转移的研究

目的 探讨尿激酶纤溶酶原激活剂（u-PA）系统的主要成份（u-PA、PAI-1及PAI-2）在口腔鳞癌组织中的表达及其与口腔鳞癌临床病理的特征。方法 应用免疫组化法检测40例口腔鳞癌和20例正常口腔粘膜组织中u-PA系统各因子的表达。结果 在正常口腔粘膜组织中u-PA系统各因子表达阴性,而在口腔鳞癌组织中u-PA系统表达则呈不均质性。在40例肿瘤中PAI-2阳性表达2例,u-PA系统中u-PA和PAI-1的表达与口腔鳞癌的病理学分级无关（P>0.05）。在21例Ⅰ、Ⅱ期口腔鳞癌中u-PA、PAI-1呈低水平表达,19例Ⅲ、Ⅳ期口腔鳞癌中则呈高水平表达,两者相比差异有显著性（P<0.05）。有颈淋巴结转移者u-PA和PAI-1呈高水平表达,而无转移者则呈低水平表达,两者相比差异有显著性（P<0.05）。u-PA与PAI-1表达水平显著呈正相关（P<0.05）。结论 u-PA系统的表达异常与口腔鳞癌的发生、发展及颈淋巴结转移有关。     

p38 MAPK在LPS诱导牙龈成纤维细胞表达uPA中作用的研究

目的：研究p38 MAPK在LPS诱导牙龈成纤维细胞表达uPA中的作用。方法：采用Western blotting观察LPS对牙龈成纤维细胞内p38 MAPK活性的影响；蛋白激酶活性实验SB203580地p38 MAPK活性的抑制作用；Northern blotting观察SB203580对LPS诱导uPA表达的影响。结果：LPS能够迅速地激活牙龈成纤维细胞内p38 MAPK的活性；SB203580能够有效地抑制牙龈成纤维细胞内的p38 MAPK的活性；经SB203580处理后，LPS对uPA的诱导作用受到显著的抑制。结论：LPS通过p38 MAPK信号转导途径诱导牙龈成纤维细胞表达uPA。     

Plasminogen activators and their inhibitors in human saliva and salivary gland tissue

The purpose of this study was to identify plasminogen activators (PA) and their specific inhibitors in human cell-free saliva and to investigate their expression in salivary gland tissue. Saliva samples were obtained from 34 patients visiting a neurological out-patient department. The activities of tissue and urokinase plasminogen activators (tPA and uPA, respectively), the relative inhibition of tPA, and the amounts of plasminogen activator inhibitors 1 and 2 (PAI-1 and PAI-2, respectively) in cell-free saliva were studied. The activities of tPA and uPA, and tPA inhibition, were measured using in-house microtiter plate assays, and PAI-1 and PAI-2 levels were measured using commercial enzyme-linked immunosorbent assay (ELISA) kits. Immunohistochemistry was used to evaluate the expression of PAs and PAIs in the salivary gland. Tissue plasminogen activator activity was found in most samples, with a mean activity of 0.63 IU ml(-1). uPA was observed in only a few samples. PAI-1 was not detected, but PAI-2 was present in all samples (with a mean value of 11.1 ng ml(-1)). The mean PAI-2 level in women was 12.4 and in men was 7.6 ng ml(-1). The activity of tPA and the relative inhibition of tPA seemed to be inversely associated. Tissue plasminogen activator, PAI-1, and PAI-2 were evident in salivary gland tissue, whereas the expression of uPA was low. The tPA activity in saliva suggests an active proteolysis. Plasminogen activator inhibitor 2 was found to be the main inhibitor of PAs in saliva.     

Immunohistochemical demonstration of the plasminogen activator system in human gingival tissues and gingival fibroblasts

The relative distribution of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2) was studied in cultured human gingival fibroblasts, healthy gingival tissues and inflamed gingival tissues by immunohistochemistry. In cultured gingival fibroblasts t-PA, u-PA and PAI-1 were expressed in cytoplasm; u-PA and PAI-1 were more intensely stained than t-PA; PAI-2 was not detectable in gingival fibroblasts. Following interleukin 1β (IL-1β) stimulation, the intensity of intracellular staining for t-PA was increased and a number of cells staining strongly for PAI-2 were seen; no difference in the intensity of immunostaining level was noted for the expression of u-PA and PAI-1 between IL-1β stimulated cells and unstimulated cells. In healthy gingival tissues, u-PA and PAI-1 displayed a wide distribution throughout all the connective tissue and epithelium; t-PA localized mainly in the connective tissue while PAI-2 showed little association with the connective tissue but did faintly stain in the epithelial layer. In inflamed gingival tissues, staining for t-PA was significantly increased in the extracellular matrix of the connective tissue, whereas staining for u-PA, PAT-I and PAI-2 was found to be slightly increased, but no significant difference was noted for staining when compared with the healthy gingival tissues. A granular distribution of t-PA, u-PA, PAI-1 and PAI-2 was noted around areas of inflammatory cell infiltration. These immunohistochemical findings indicate that the plasminogen activator system produced by fibroblasts may be influenced by the presence of the inflammatory mediator IL-1β. In addition, the significant increase of t-PA in inflamed connective tissue and the wide expression of these components around inflamed cells may contribute to connective tissue degradation and may relate to the migration and localization of monocytes/macrophages in inflamed tissue.     

Immunohistochemical localization of tissue-type plasminogen activator and type I plasminogen activator inhibitor in radicular cysts

BACKGROUND: The plasminogen/plasmin proteolytic system participates in a wide variety of extracellular matrix degradation. Detailed knowledge of plasminogen activators (PAs) and their inhibitors may be important for understanding the pathogenesis of radicular cysts. The purpose of this study was to investigate the in situ localization of tissue-type PA (t-PA) and type I PA inhibitor (PAI-1) in radicular cysts. METHODS: Thirty formalin-fixed, paraffin-embedded specimens of radicular cysts were examined using immunohistochemistry. In addition, another section from each radicular cyst specimen was stained with hematoxylin and eosin to assess the presence of inflammatory infiltrates. Differences in t-PA and PAI-1 expression between tissues with low and high levels of inflammation were subsequently analyzed using Fisher's exact test. RESULTS: Both t-PA- and PAI-1-positive cells were detected in the lining epithelium, connective tissue, inflammatory infiltrates, and endothelium. In addition, the t-PA signal was mainly expressed in epithelial cells. However, the PAI-1 signal was mainly expressed in fibroblasts. Moreover, significantly greater t-PA as well as PAI-1 expression was noted in radicular cysts with high levels of inflammation as compared to tissues with low levels of inflammatory cell infiltrates (P < 0.05). CONCLUSIONS: The present study confirms earlier indications of local production of PA and its inhibitor in radicular cysts. In addition, this study further shows the tissue localization of the antigens for t-PA as well as PAI-1, and demonstrates that the expression of both t-PA and PAI-1 increases with the grade of inflammation in radicular cysts.     

兔关节盘前移位后髁突软骨PRG4 mRNA表达改变的研究

目的 分析兔颞颌关节盘前移位后髁突软骨PRG4 mRNA表达的动态改变及其意义。方法 62只日本大耳白兔,48只建立关节盘前移位的动物模型,分期处死。6只为手术对照组,8只为空白对照。合成兔PRG mRNA探针,原位杂交检测关节盘前移位后髁突软骨PRG4 mRNA表达的动态改变。结果 PRG4 mRNA表达于髁突全层软骨。关节盘前移位后,PRG4 mRNA表达一过性升高,8周后回落至正常。4只样本术区骨关节病样表现,软骨崩解区域PRG4 mRNA的表达水平显著下降。结论 关节盘前移位后,髁突软骨细胞PRG4 mRNA一过性升高以保护髁突软骨表层的完整性,促进滑液润滑,有利于实现关节的适应性改建。?     

口腔鳞癌中u—PA系统的表达与颈淋巴结转移关系的研究

目的 探讨u－PA系统的主要成分（u－PA、PA1－1及PAI－2）在口腔鳞癌组织中的表达及其与口腔临床病理特征的关系。方法 应用免疫组化法检测了40例口腔鳞癌和20例正常口腔黏膜组织中u－PA系统各因子的表达。结果 在正常口腔黏膜组织中u－PA系统各因子表达阴性,而在口腔鳞癌组织中u－PA系统表达则呈不均质性;在40例肿瘤中PAI－2阳性表达2例,u－PA系统u－PA和PAI－1的表达与口腔鳞癌的病理学分级无关（P＞0．05）;在21例Ⅰ、Ⅱ期口腔鳞癌中u－PA、PAI－1呈低水平表达,19例Ⅲ、Ⅳ期口腔鳞癌中则呈高水平表达,两者相比差异有显著性（P＜0．05）;在有颈淋巴结转移组中u－PA系统和PAI－1呈高水平,而在无转移组中则呈低水平表达,两者相比差异有显著性（P＜0．05）;且u－PA与PAI－1表达水平呈显著正相关（P＜0．05）。结论 u－PA系统的表达异常可能与口腔鳞癌的发生发展及颈淋巴结转移有关。     

颞下颌关节紊乱病关节液中尿纤溶酶原激活物及其受体的表达

目的 检测颞下颌关节(TMJ)关节液中尿纤溶酶原激活物(urokinase-type plasminogen activator,uPA)及受体(urokinase-type plasminogen activator receptor,uPAR)的分泌量,探讨TMJ液中uPA及uPAR与颞下颌关节紊乱病(TMD)的关系.方法 采用酶联免疫吸附实验法检测56例TMD患者的64侧关节和10名健康志愿者的16侧关节的关节液标本中的uPA及uPAR的量.将符合纳入标准的48侧TMD患者的关节液标本根据临床诊断分为关节炎性组(A组)、结构紊乱组(B组)、骨关节病组(C组),每组16侧;10名健康志愿者的16侧关节液设为对照组(D组).结果 TMD中A组、B组、C组、D组uPA的检出量分别为(51.200±8.786)ng/L、(53.667±11.894)ng/L、(81.278±25.828)ng/L、(17.960±9.859)ng/L;uPAR检出量分别为(5.840±0.179)ng/L、(6.168±1.465)ng/L、(2.416±0.525)ng/L、(2.416±0.525)ng/L.uPA和uPAR表达量均高于健康对照组(P＜0.05),C组uPA和uPAR表达量高于A组及B组(P＜0.05),但A组与B组差异无统计学意义(P＞0.05).结论 TMJ内过度分泌的uPA和uPAR可能参与了TMD关节组织的病理破坏过程;关节液中uPA和uPAR的水平可部分反映TMD病变的程度,可作为反映TMD关节损害和代谢的客观生化分子标志.     

Association of gene polymorphisms for plasminogen activators with alveolar bone loss

BACKGROUND AND OBJECTIVE: The plasminogen activating system is a protease/inhibitor system central to extracellular matrix remodeling with a suggested role in periodontal disease pathology. A few studies have reported polymorphisms in the genes of plasminogen activator inhibitors to be associated with periodontal disease severity. Two gene polymorphisms - a BamHI restriction fragment length polymorphism in the urokinase plasminogen activator gene (uPA) and a HindIII restriction fragment length polymorphism in the plasminogen activator inhibitor type 1 gene (PAI-1) - have been associated with conditions having a vascular component, and our objective was to assess the association of these gene polymorphisms with alveolar bone loss in chronic periodontal disease of adults. MATERIAL AND METHODS: Genotype was determined by polymerase chain reaction amplification of whole blood, pertinent histories were obtained by interview, and alveolar bone loss was assessed from current radiographs. RESULTS: In 77 elderly patients with a normal distribution of alveolar bone loss, we demonstrated a significant association between levels of alveolar bone loss and these polymorphisms in the uPA and PAI-1 genes. Controlling for the contributions of smoking or diabetes to periodontal bone loss, estimated odds ratios for predicting lower levels of alveolar bone loss, associated with a greater degree of periodontal health, were strongest when defined by the concurrent presence of a homozygous urokinase plasminogen activator genotype and the nuclease-sensitive plasminogen activator inhibitor type 1 (HindIII) allele (odds ratio = 2.6; 95% confidence interval: 5.8-1.3). CONCLUSION: The urokinase plasminogen activator (BamHI) and plasminogen activator inhibitor type 1 (HindIII) genotypes may serve as useful markers for heritability of bone loss associated with periodontal disease.