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口腔黏膜下纤维性变(oral submucous fibrosis,OSF)是一种慢性、隐匿性、具有癌变倾向的口腔黏膜疾病。其主要临床表现为口腔黏膜纤维化、进食刺激性食物时口内疼痛,严重者可致张口和进食困难。目前,OSF的诊断手段主要包括临床检查和病理诊断。本文旨在结合笔者的临床经验和最新的相关文献,评述OSF诊断方法的研究进展,如分子生物学方法在OSF诊断中的应用、OSF合并症的诊断等。     

HGF/c-met在口腔黏膜下纤维化组织中表达的研究

收集早、中、晚期口腔黏膜下纤维性变(oral submucous fibrosis,OSF)组织各10例及正常的口腔颊黏膜组织5例,采用免疫组化SABC法对各种组织中的HGF及c-met的表达情况进行检测。结果显示在正常的口腔颊黏膜组织中HGF和c-met呈阳性表达,早、中、晚期OSF组织中表达明显降低(P<0.05)。说明HGF/c-met在OSF组织中表达受到抑制,促进了OSF的发病。     

口腔黏膜下纤维化中VEGF mRNA的表达研究

目的:研究口腔黏膜下纤维化(oral submucous fibrosis,OSF)患者早、中、晚期组中血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达,探讨VEGF与OSF微血管病变发生发展的关系.方法:选取OSF患者30例,其中早、中、晚期组各10例,5例健康志愿者为对照组.每例研究对象取其1:3腔黏膜,运用逆转录聚合酶链反应(RT-PCR)方法,研究OSF患者早、中、晚期组中VEGF mRNA的表达水平.结果:VEGF mRNA在OSF早、中期的表达均较正常口腔黏膜高(P＜0.05):VEGF mRNA在OSF早期的表达均明显高于中、晚期(P＜0.05),其中中期的表达与晚期相比差异无显著性(P＞0.05).结论:VEGF对OSF微血管病变的影响主要在黏膜下层,各种调控因素在基因转录水平导致VEGF的高表达可能是OSF局部组织缺血的代偿.     

Wnt1在口腔黏膜下纤维性变中的表达

目的:研究wnt1在口腔黏膜下纤维性变(OSF)患者治疗前后的表达并探讨其在OSF发生发展中的作用.方法:40例OSF中期患者采用曲安奈德+丹参局部封闭治疗4周,治疗前及第5周复诊时记录VAS及张口度,取唾液及龈沟液ELISA检测wnt1表达,实时荧光定量PCR检测OSF病变颊黏膜中wnt1 mRNA表达.结果:治疗前OSF组wnt1表达[颊黏膜定量PCR(36.89±10.40)×10-5,唾液浓度(61.61 ±4.45)ng/L,龈沟液浓度(56.20±3.65) ng/L]均高于正常组[颊黏膜定量PCR(4.63±1.53)×10-5,唾液浓度(40.26 ±3.00) ng/L,龈沟液浓度(53.45±1.74) ng/L] (P＜0.01);OSF组经治疗后,唾液与龈沟液中wnt1表达均降低,且与OSF临床严重程度呈正相关(P＜0.05).结论:wnt1可能参与OSF的发生发展,其在唾液与龈沟液中的实时监测有望成为评价OSF诊疗的无创性检控手段之一.     

The fibroblast population in oral submucous fibrosis

The purpose of the investigation was to compare the morphology of fibroblasts cultured from healthy oral mucosa and mucosa of patients with oral submucous fibrosis (OSF) and to collate the occurrence of cell types of similar morphology. Cells cultured from biopsy specimens from the buccal mucosa of six subjects who did not chew the areca nut and six patients with OSF who chewed areca nut were grown according to standard techniques. Ninety cells per cell line were recorded daily for 8 days, classified into types F1, F2 and F3 according to their morphology, and the results statistically analyzed. We found that there was a relative increase of F3 cells in relation to Fl cells in OSF resulting in the ratio of F3 to F1 cells being significantly larger in OSF than the ratio in the controls. As it has been reported that F3 cells m rat connective tissues produce significantly more collagen types I and III than F1 cells, we concluded that a change of fibroblast population has occurred in OSF and that this relative increase of F3 cells in humans, which could be committed to the production of large quantities of collagen, can be an explanation for the excessive collagen formation in OSF.     

婴幼儿脉管性疾病外周血和瘤内血bFGF的表达及意义

目的:通过检测婴幼儿血管性疾病外周静脉血(简称外周血)、瘤内血中的碱性成纤维细胞生长因子(bFGF)的表达,探讨对血管瘤和血管畸形进行鉴别诊断的可能.方法:应用酶联免疫吸附法(ELISA)检测资料完整的14例静脉畸形患儿瘤内血和外周血bFGF,检测49例血管畸形患儿、32例血管瘤患儿和23例对照婴幼儿外周血清中bFGF的浓度,采用SPSS11.5软件包对数据进行t检验和方差分析.结果:静脉畸形瘤内、外周血清中bFGF浓度有显著差异(P＜0.05),瘤内血清bFGF浓度高于外周血清:血管瘤、血管畸形和对照组外周血清bFGF浓度无统计学差异(P＞0.05).结论:静脉畸形患儿瘤内血清的bFGF浓度高于外周血清,检测外周血bFGF,不能鉴别血管瘤和血管畸形.     

毛细血管瘤组织中成纤维细胞生长因子（bFGF）的表达水平

成纤维细胞生长因子（bFGF）是一组结构相关的多肽类血管形成因子，它能诱导或促进新生毛细血管的形成，对血管瘤的发展有关键作用。本文通过免疫组织化学方法，观察bFGF在颌面部毛细血管瘤不同分期的表达变化，初步探讨它在肿瘤增殖退化病理演变过程中的作用。     

成纤维细胞生长因子对兔下颌牵张成骨的影响

目的 研究局部应用碱性成纤维细胞生长因子（bFGF)对兔下颌牵引张成骨的影响。方法 成年大耳白兔8只，随机分为A、B两组，每组4只。用自行研制的牵张器延长双侧下颌骨6mm，用bFGF（20ng/ml）注入A组动物的牵张区，不注射bFGF的B组动物作为对照。在牵张结束后4周处死所有动物，取双侧下颌骨标本进行X线和组织学检查。结果 组织学分析结果显示，局部给予bFGF的A组动物下颌牵张后新骨生成速度和数量优于B组动物。结论 外源性导入碱性成纤维细胞生长因了可能有促进下颌牵引张成骨的作用。     

PTTG和bFGF在口腔鳞癌中的表达及意义

目的：探讨垂体肿瘤转化基因（pituitary tumor transforming gene，PTFG）和碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）在口腔鳞癌中的表达及相互关系，研究它们的表达与肿瘤临床病理指标的联系。方法：应用SP染色法检测PTTG蛋白和bFGF在55例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率。结果：在口腔鳞癌中PTTG和bFGF的阳性表达率分别为78．2％和67．3％，其阳性率及表达等级均显著高于正常对照组（P〈0．05）。PTTG在中一低分化组和有淋巴结转移组中的表达显著高于高分化组和无淋巴结转移组（P〈0．05）。PTTG表达与bFGF表达成等级正相关（r=0．382，P〈0．05）。结论：PTFG或bFGF与口腔鳞癌生物学行为及预后有密切关系，二者的联合检测，有助于口腔鳞癌恶性程度和预后的判断。     

碱性成纤维细胞生长因子在人牙胚中表达的免疫组化定位

目的：观察碱性成纤维细胞生长因子（ｂＦＧＦ）表达与人牙胚发育和分化的关系。方法：采用免疫组化方法观察ｂＦＧＦ在人牙胚发育中的定位。结果：钟状期牙胚中前成釉细胞为ｂＦＧＦ强阳性，星网层可疑；成牙本质细胞为强阳性，牙乳头组织中阳性分布不均，靠近成牙本质细胞的牙乳头细胞为阳性；牙囊细胞阳性，血管内皮细胞呈强阳性；骨组织为阴性。结论：ｂＦＧＦ与成釉细胞、成牙本质细胞的分化和成熟有关。     

c-fos、c-jun蛋白在口腔黏膜下纤维性变组织中的表达研究

目的 :检测c -fos、c -jun蛋白在口腔黏膜下纤维性变 (OSF)组织中的表达及分布 ,探讨其在OSF发病机制中的作用。方法 :采用免疫组化SABC法 ,用c -fos、c -jun兔抗人多克隆抗体检测 5 6例OSF病变组织、15例口腔黏膜扁平苔藓 (OLP)病变组织及 10例正常口腔黏膜组织中的c -fos、c -jun蛋白的表达及分布。 结果 :c -fos、c -jun蛋白在OSF病变组织有明显的阳性表达 ,正常口腔黏膜及OLP组织为阴性表达 (P <0 .0 5 )。其表达主要位于OSF病变上皮组织 (P <0 .0 5 ) ,且表达强度为正相关 (γs=0 .744 ,P <0 .0 1)。结论 :c -fos、c -jun蛋白在OSF病变组织中高表达 ,在OSF发病机制中有重要作用。     

口腔鳞癌组织中bFGF的表达与血管生成的关系

目的:观察口腔鳞癌组织中碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)的表达及其与微血管密度(microvesseldensity,MVD)之间的关系,探讨bFGF对口腔鳞癌血管生成的作用及意义。方法:收集口腔鳞癌标本42例(其中有淋巴结转移者14例)和10例正常口腔黏膜组织,用免疫组织化学染色法探测bFGF和CD34的表达情况并计数微血管密度(MVD)。结果:口腔鳞癌中bFGF表达阳性率为69. 05% (29 /42),显著高于正常口腔组织30% (3 /10),口腔鳞癌中MVD显著高于正常组织,且随病理分化不良而增高(P< 0. 05),有淋巴结转移组MVD显著高于无淋巴结转移组(P< 0. 05),bFGF表达阳性组中MVD明显高于bFGF表达阴性组(P< 0. 05 ),bFGF的表达与MVD呈正相关(P< 0. 01 )。结论:bFGF在口腔鳞癌组织中的高度表达在口腔鳞癌发生发展中起重要作用,并与其血管生成和淋巴结转移有关。     

Regeneration of periodontal tissues by basic fibroblast growth factor

Several growth factors (or cytokines) have recently received attention because of their ability to actively regulate various cellular functions of periodontal ligament (PDL) cells and the effects of topical application of such factor(s) on periodontal tissue regeneration has been evaluated. In this study, we examined the role of basic fibroblast growth factor (bFGF) in the wound healing and regeneration of periodontal tissues. Alveolar bone defects (such as 2-wall, 3-wall and furcation class II bone defects) were created surgically in beagle dogs and primates. Recombinant bFGF was topically applied to the artificial bony defects. Six or 8 wk after application, the periodontal regeneration was morphologically and histomorphometrically analyzed. In all sites where bFGF was applied, significant periodontal ligament formation with new cementum deposits and new bone formation was observed in amounts greater than in the control sites. We found it noteworthy that no instances of epithelial down growth, ankylosis or root resorption were observed in the bFGF sites. In vitro studies demonstrated that bFGF enhances the proliferative responses of human PDL cells, which express FGF receptor-1 and -2, but inhibits the induction of alkaline phosphatase activity and mineralized nodule formation by PDL cells. Interestingly, we observed that the mRNA level of laminin in PDL cells, which plays an important role in angiogenesis, was specifically upregulated by bFGF stimulation, but that of type I collagen was downregulated. The present study demonstrates that bFGF can be applied as one of the therapeutic modalities which actively induce periodontal tissue regeneration. The results of in vitro studies suggest that by suppressing the cytodifferentiation of PDL cells into mineralized tissue forming cells, bFGF may play important roles in wound healing by promoting angiogenesis and inducing the growth of immature PDL cells, and may in turn accelerate periodontal regeneration.     

NGF、bFGF对体外培养人牙髓细胞增殖与分化的作用

目的:研究NGF、bFGF及两者联合应用对体外培养人牙髓细胞(HDPC)的增殖与分化作用的影响。方法:用MTT和ALP活性检测法,观察对照组(10ml/L FBS的DMEM培养液)与实验组(10U/ml NGF、10μg/L bFGF、10U/ml NGF 10μg/L bFGF、5U/ml NGF 5μg/L bFGF、1U/ml NGF 1μg/L bFGF)对体外培养的第5~8代HDPC增殖与分化作用的影响。对实验数据行Dunnett-t检验。结果:与对照组相比,10U/ml的NGF可显著促进HDPC的增殖(P<0.05);对ALP的活性没有明显的增强作用(P>0.05);10μg/LbFGF可显著促进HDPC的增殖(P<0.05),但对HDPC的ALP活性,却有一定的抑制作用(P>0.05);10U/ml NGF 10μg/L bFGF和5U/ml NGF 5μg/L bFGF既可显著地促进HDPC增殖(P<0.05),又可增强HDPC的ALP活性(P<0.01)。结论:一定浓度的NGF与bFGF组合既可促进HDPC增殖,又可促进其分化,两者对HDPC有显著的功能放大性协同增强作用。     

Alterations in expression of retinoid receptor beta and p53 in oral submucous fibrosis

OBJECTIVE: Knowledge of the molecular pathogenesis of oral submucous fibrosis (OSF), a potentially malignant condition with high risk of transition to oral cancer, is meagre. Alterations in the expression of retinoic acid receptor beta (RARbeta) and tumor suppressor gene, p53 are early events in oral tumorigenesis. The aim of this study was to investigate the alterations in the expression of RARbeta and p53 in OSF lesions and determine their association with disease pathogenesis. METHODS: The expression of RARbeta and p53 proteins was analyzed by immunohistochemistry in 50 cases of OSF and 30 histologically normal oral tissues. RESULTS: No detectable RARbeta expression was observed in 35 of 50 (70%) OSF cases. p53 protein accumulation was observed in 24 of 50 (48%) OSF cases analyzed. Thirty-six percent OSF lesions showed loss of RARbeta and p53 overexpression. Interestingly, 41 of 50 (82%) of OSF lesions showed altered expression of at least one of these two proteins. CONCLUSION: Altered expression of either RARbeta or p53 in majority of OSF lesions suggests their association with disease pathogenesis and warrants follow-up to determine whether OSF lesions harboring concomitant alterations in RARbeta and p53 are at a high risk of transition to malignancy.     

碱性成纤维细胞生长因子对体外大鼠牙胚分化的影响

目的：观察外源性ｂＦＧＦ对体外培养牙胚发育和分化的作用。方法：采用体外培养１７ｄＳＤ胎鼠第一磨牙牙胚，培养液中加入１０ｎｇ／ｍｌ人重组ｂＦＧＦ（ｈｂＦＧＦ），分别培养３、６、９ｄ，组织学观察。结果：外源性ｈｂＦＧＦ可使大鼠牙胚的细胞分化和牙本质基质分泌加快。结论：外源性ｂＦＧＦ可以促进大鼠牙胚的发育，加速牙齿发育。     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10-¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodiflerentiation of PDL cells into mineralized tissue forming cells.     

Epidemiological survey of oral submucous fibrosis and leukoplakia in aborigines of Taiwan

A population-based survey was designed to investigate the prevalence of areca/betel quid chewing, oral submucous fibrosis and leukoplakia in a typical aboriginal community of southern Taiwan. Three hundred and twelve people 20 years of age or older were collected in the study. The prevalence of chewing areca/betel quid was 69.5%, with an average of 17.3 portions a day for an average 24.4 years. More women (78.7%) than men (60.6%) chewed areca/betel quid. The prevalences of oral submucous fibrosis and leukoplakia were 17.6% and 24.4%, respectively. It was found that the odds ratio for chewing areca/betel quid and having at least one of the above oral mucosal lesions was 8.21. Any additional smoking or drinking habits were not significant for having oral mucosal lesions. Although the areca/betel quid in Taiwan does not contain any tobacco, a significant association was still identified between areca/betel quid chewing and oral mucosal lesions.     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10⁻¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodifferentiation of PDL cells into mineralized tissue forming cells.     

碱性成纤维细胞生长因子促进颌骨缺损修复的研究

目的 :评价碱性成纤维细胞生长因子 (bFGF)对成骨的促进作用 ,以期为颌骨缺损提供一种较为理想的替代材料。方法 :18只成年兔双侧下颌骨下缘各造成 15mm× 6mm的全层骨膜骨质缺损 ,每一缺损作为一个实验单位 ,按自身同期配对设计 ,分别植入复合骨和天然型无机骨 (NNB)。术后 3、6、12周取下标本 ,进行甲苯胺蓝染色观察、四环素荧光检查以及组织学成骨面积定量分析。结果 :复合骨新骨形成及钙化早 ,同期成骨面积大于NNB(P <0 .0 1)。结论 :合适的复合条件下bFGF可有效地促进软骨及骨的生长 ,复合骨新骨形成及钙化早 ,同期成骨量多 ,修复效果优于NNB。