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目的 探讨牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)W83、ATCC33277刺激人牙周膜成纤维细胞(HPDLFs)分泌基质金属蛋白酶-1(MMP-1)、金属蛋白酶组织抑制剂-1(TIMP-1)的变化.方法 本研究于2010年9月至2011年6月在中国医科大学口腔医学院中心实验室进行.将P.gingivalis W83、ATCC33277作用于HPDLFs0、6、12、24、48h后,运用酶联免疫吸附法(ELISA)检测细胞上清液中MMP-1、TIMP-1质量浓度变化,并计算MMP-1/TIMP-1值.结果 P.gingivalis感染HPDLFs的MMP-1和TIMP-1表达均增强,并呈时间依赖性;且MMP-1/TIMP-1值明显高于未感染的对照组(P＜0.05).P.gingivalis W83感染后MMP-1/TIMP-1值明显高于P.gingivalis ATCC33277(P＜ 0.05).结论 P.gingivalis具有促进HPDLFs分泌MMP-1、TIMP-1的作用,且可造成牙周组织破坏;P.gingivalis W83降解细胞外基质的能力高于P.gingivalisATCC33277.     

牙龈卟啉单胞菌侵入对血管内皮细胞E选择素表达的影响

目的：观察牙龈卟啉单胞菌（P.g）ATCC33277和W83侵入对血管内皮细胞E选择素转录和翻译的影响。方法：建立体外P.g侵入血管内皮细胞模型,反转录-聚合酶链式反应检测E选择素基因表达;蛋白质印迹技术检测E选择素膜蛋白表达的变化;采用SAS8.12软件包,多组均数之间的比较采用重复测量资料的方差分析,P〈0.05为差异有统计学意义。结果：P.gATCC33277和W83侵入后4h即上调E选择素基因表达（P〈0.05）,8h显著回落（P〈0.05）,24h恢复至基线水平;E选择素基因在P.gATCC33277/W83侵入后4h、8h、24h和TNFα处理0.5h后的相对表达水平分别为对照组的294.0%/326.4%、179.3%/224.6%、126.7%/128.3%和365.5%。P.g侵入后4h即诱导E选择素蛋白表达（P〈0.05）,8h达高峰（P〈0.05）,24h有持续高表达（P〈0.05）;E选择素蛋白相对表达水平分别为对照组的295.4%/343.6%、386.3%/394.3%、238.7%/296.4%和413.8%。P.gW83诱导血管内皮细胞E选择素表达的能力强于ATCC33277（P〈0.05）。结论：P.g侵入可造成血管内皮细胞E选择素高表达的动脉粥样硬化样改变,在动脉粥样硬化炎症病理反应中有重要意义。     

牙龈卟啉单胞菌影响血管内皮细胞增殖和凋亡的实验研究

目的 观察牙龈卟啉单胞菌(Porphyromonas gingivatis,Pg)对血管内皮细胞增殖和凋亡的影响,探讨Pg在动脉粥样硬化发病机制中的作用.方法 建立体外Pg侵入血管内皮细胞模型,甲基噻唑基四唑(MTT)法观察细胞增殖,碘化丙啶染色、流式细胞仪分析细胞周期,Annexin-V-FITC凋亡试剂盒检测细胞凋亡.结果 PgATCC33277侵入后72 h细胞增殖活性降低12.46%,Pg W83侵入后72 h细胞增殖活性降低10.47%(F=786.68,P<0.01);Pg W83侵入后24 h使G1期细胞增加(F=43.23,P<0.01),ATCC 33277侵入后48 h使G1期细胞增加(F=66.72,P<0.01);Pg侵入后24 h即诱导细胞凋亡(F=1074.56,P<0.01).结论 Pg可能通过细胞毒性及诱导凋亡作用,加重血管内皮细胞的局部炎性反应,在动脉粥样硬化炎性病理反应中有重要意义.     

牙龈卟啉单胞菌外膜囊泡差异表达mRNA的生物信息学分析

目的 探讨牙龈卟啉单胞菌(P.gingivalis)W83与ATCC33277外膜囊泡mRNA表达谱的差异.方法 超速离心法分离W83和ATCC33277来源的外膜囊泡,进行粒径检测、电镜及Western blot鉴定;高通量测序检测W83和ATCC33277外膜囊泡mRNA表达谱,筛选差异表达的mRNA,进行基因本体...     

牙龈卟啉单胞菌感染MG63细胞对细胞周期的影响

目的:通过体外建立牙龈卟啉单胞菌(P. gingivalis)感染MG63细胞模型,探讨P. gingivalis ATCC 33277菌株感染MG63细胞对细胞周期的影响及发生机制。方法:P. gingivalis ATCC 33277菌株感染MG63细胞,MTT法检测细胞的增殖,流式细胞仪检测细胞周期的分布,实时定量PCR和蛋白印记方法检测Cyclin D、Cyclin E的表达。结果:MOI为100∶1,P. gingivalis ATCC 33277菌株感染MG63细胞,能够抑制细胞增殖,同时导致细胞在G1期发生停滞,Cyclin D、Cyclin E基因和蛋白在24 h、48 h均有明显下降,与对照组相比有统计学差异。结论:牙龈卟啉单胞菌感染MG63细胞Cyclin D、Cyclin E基因和蛋白表达下调,抑制细胞的增殖,导致细胞停滞在G1期。     

两种fimA基因型牙龈卟啉单胞菌刺激下口腔上皮细胞ICAM-1的表达

目的比较2种fimA基因型牙龈卟啉单胞菌（Porphyromonas gingivalis,P．gingivalis）刺激下口腔上皮细胞ICAM-1的表达水平。方法实验组用P．gingivalis ATCC 33277（Ⅰ型菌毛组）,W83（Ⅳ型菌毛组）,47A-1（Ⅳ型菌毛组）分别与KB细胞（ATCC CCL17）共同孵育24h;对照组为未受P.gingivalis刺激的KB细胞。分别在1h、3h、6h和24h收集细胞,运用流式细胞仪检测KB细胞膜上ICAM-1的动态表达。结果P．gingivalis刺激细胞后3h、6h和24h,实验组ICAM-1的表达水平均高于对照组;2种fimA基因型P．gingivalis调节KB细胞表达ICAM-1的方式相似,Ⅳ型菌毛组的调节作用强于Ⅰ型菌毛组。结论P.gingivalis上调口腔上皮细胞表达ICAM-1的水平与其fimA基因型相关,提示P.gingivalis致病性与其fimA基因型相关。     

不同fimA基因型牙龈卟啉单胞菌刺激口腔上皮细胞白细胞介素-8的表达

目的比较不同fimA基因型牙龈卟啉单胞菌（P.gingivalis）刺激下口腔上皮细胞白细胞介素-8（IL-8）的表达水平,初步探讨P.gingivalis致病性与其fimA基因型的关系。方法P.gingivalis ATCC 33277（Ⅰ型fimA）、W83、47A-1（Ⅳ型fimA）和KB细胞ATCC CCL-17共同孵育24 h,以未受刺激的KB细胞作为对照组,分别在1、3、6、24 h收集细胞和培养上清液。RT-PCR检测KB细胞IL-8 mRNA的动态表达,酶联免疫反应检测培养上清液中IL-8的动态变化。结果2种fimA基因型菌株刺激1 h,KB细胞IL-8 mRNA的表达上调至峰值,高于对照组,3~24 h IL-8 mRNA的表达水平接近对照组;P.gingivalis感染细胞后3~24 h,上清液中的IL-8水平低于对照组,Ⅳ型菌株低于Ⅰ型菌株;IL-8 mRNA及其蛋白表达不完全一致,提示IL-8的表达存在转录后水平的调节。结论fimA基因型与口腔上皮细胞IL-8的表达水平相关,提示P.gingivalis致病性与其fimA基因型相关。     

牙龈卟啉单胞菌入侵血管内皮细胞的实验研究

目的:体外观察牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖的影响,观察Pg对HUVEC的入侵能力,从而探讨Pg对内皮细胞功能损伤的途径,以期为牙周病在动脉粥样硬化(Atherosclerosis,As)发病机制中的作用提供依据。方法:用感染复数(multiplicity of infection,MOI)1:10,1:100PgATCC33277分别干预HUVEC 4h、8h、12h、24h,未受Pg ATCC33277干预的HUVEC作为阴性对照组,倒置显微镜下观察HUVEC形态,CCK-8法测定HUVEC细胞的增殖情况,透射电镜下观察Pg ATCC33277对HUVEC的入侵情况。结果:在24h内,与对照组相比,受Pg ATCC33277感染的HUVEC的细胞形态未见明显影响,仍呈典型的"铺路石"单层贴壁生长,Pg ATCC33277对HUVEC细胞增殖未见明显影响,透射电镜下观察发现Pg ATCC33277可以黏附HUVEC细胞膜表面,入侵HUVEC,直接定植或以空泡的形式存在于细胞的胞质中。结论:Pg可以入侵内皮细胞,以在细胞内定植的方式逃避宿主的防御反应,内皮细胞的形态和增殖未见明显改变,从而形成第二次慢性感染过程,进一步影响内皮细胞正常功能的发挥。     

多粘菌素B和热处理对牙龈卟啉单胞菌诱导ECHUV产生sICAM-1的影响

目的：观察多粘菌素B和热处理对牙龈卟啉菌诱导人类脐静脉血管内皮细胞（endothelial cells of human umbilical vein,ECHUV）产生可溶性细胞间粘附分子－1（soluble intercellular adhesion molecule-1,sICAM-1）的影响，以探讨P．gingivalis诱导ECHUV产生sICAM-1的有效部位。方法：采用酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）测定培养上清中sICAM-1的含量。结果：P.gingivalis、大肠杆菌脂多糖（LPS）和肿瘤坏死因子－α（TNF－α）强烈诱导ECHUV产生sICAM-1，多粘菌素B明显抑制大肠杆菌LPS的诱导作用，热处理完全废除了TNF－α的作用，而这两种方法都不能降低P.gingivalis对ECHUV产生sICAM-1的影响。另外，P.gingivalis毒力株W83的诱导强度明显高于非毒力株ATCC33277。结论：P.gingivalis可能通过LPS以外的耐热的有效位点诱导ECHUV产生sICAM-1，这一位点可能与P.gingivalis的有效毒力因子有关。     

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目的探讨核转录因子一KB（NF—KB）／抑制因子KB（IKB）信号通路在调控牙龈卟啉单胞菌（Pgingivalis）感染诱导血管内皮细胞细胞间黏附分子-1（ICAM-1）高表达过程中的作用。方法本研究于2008年3月至2011年1月在中国医科大学中心实验室及中国医科大学口腔医学院中心实验室进行。已知P．gingivalis感染血管内皮细胞可诱导其ICAM-1基因和蛋白表达增高。采用Real-timePCR和Westernblot法检测P．gingivalis感染后血管内皮细胞ICAM-1mRNA和蛋白表达;应用5、10、20μmol／L的蛋白酶体抑制剂MGl32分别预处理血管内皮细胞15、30、60min,再将细胞与P．gingivalis共同培养8h,观察细胞ICAM-1mRNA和蛋白表达的变化,分析NF—KB／IKB信号通路对ICAM-1表达的调控作用。结果10txmol／LMGl32预处理30rain,Pgirtgivalis感染的血管内皮细胞ICAM—lmRNA表达具有下降趋势;20p~mol／LMGl32预处理15min,P．gingivalis感染的血管内皮细胞ICAM-1表达即开始下降,预处理时间延长至30、60min,ICAM-1表达下降更趋明显。结论阻断NF—KB／IKB信号通路可抑制P．gingivalis感染诱导的血管内皮细胞ICAM-1表达。     

Pediatric vascular lesions

The seminal work of Mulliken and Glowacki in 1982 elucidated the histological differences between hemangiomas and vascular malformations: the former are characterized by endothelial cell proliferation, whereas the latter contain mature endothelial cells. Hemangiomas proliferate and then involute, whereas malformations remain stable in size, growing proportionally with the child. Vascular malformations are classified by the predominant vessel type within the lesion (capillary, venous, arterial, and lymphatic). Histological classification therefore correlates with clinical behavior. Treatment of hemangiomas is generally conservative; however, intervention may be required as a result of cosmetically concerning, function-threatening (e.g., interference with eyesight), or life-threatening (e.g., airway obstruction) lesions. Options include steroid therapy, laser treatment, and/or surgical excision. Vascular malformations do not involute and are more likely to require treatment. Treatment options include embolization and surgical resection. Understanding the clinical course of pediatric vascular lesions allows the surgeon to find an appropriate balance between watchful observation, providing reassurance when appropriate, and intervention when needed.     

Mandibular vascular leiomyoma

A 54-year-old man had a symptomless swelling on the buccal aspect of the right mandibular molar area, this was associated with a multilocular radiolucency. Histologic and ultrastructural examination of the lesion revealed a vascular leiomyoma. This is only the fourth intrabony leiomyoma of the jaws to be reported.     

Facial developmental vascular anomalies

Developmental vascular anomalies of the head and neck may give rise to profound esthetic problems that are a challenge to treat. These vascular anomalies may also be of significance in dentistry, particularly where extractions have to be carried out This brief report describes a number of children who presented with different variations of these anomalies.     

Etiopathogenesis of vascular anomalies

Vascular malformations are localized errors of vascular development. They are often identified on the skin as "birthmarks" of various sizes and shapes.     

Detection of vascular endothelial growth factor/ vascular permeability factor in periapical lesions

Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF), is a multifunctional cytokine. It is overexpressed in several conditions, which are characterized by vascular hyperpermeability and angiogenesis. In this investigation, we have evaluated the possibility that VEGF/VPF could be expressed in periapical lesions. We studied 17 periapical granulomas and 6 periapical cysts by immunohistochemistry. An immunopositive reaction for VEGF/VPF was observed in all 23 periapical lesions; however, the intensity of immunostaining by anti-VEGF antibody varied according to histopathological findings. In periapical granulomas without epithelium, almost all of the inflammatory cells were immunoreactive to anti-VEGF/VIP antibody. In periapical granulomas, which had rests of Malassez in them, some inflammatory cells were stained. On the other hand, epithelial cells always were stained by VEGF/VPF antibody, both in periapical lesions with epithelium and in radicular cysts. This study demonstrated that periapical lesions express VEGF/VPF, although with some differences in cell immunolabeling, which correlated to the lesions' stages of development. Initially, VEGF/VPF would assure angiogenesis and vascular hyperpermeability, resulting in accumulation of inflammatory cells, later it could be involved in cyst fluid accumulation. We hypothesize, therefore, that VEGF/VPF expression plays an important role in the pathogenesis of periapical granulomas and enlargement of radicular cysts by several mechanisms.     

Current concepts of vascular anomalies

The field of vascular anomalies is confusing because numerous types of lesions exist, different anomalies often look similar, and imprecise terminology commonly is used. Pharmacotherapy is effective for certain vascular tumors; sclerotherapy generally is the primary treatment of problematic lymphatic and venous malformations. Arteriovenous malformations remain difficult to manage because of their high progression and recurrence rates. Propranolol has gained popularity recently for the treatment of problematic infantile hemangioma, but its efficacy and safety compared with corticosteroid therapy have not been studied. Continuing education is needed to increase the use of accepted biologic terms to describe vascular anomalies; this will improve patient care and facilitate research. As vascular anomalies centers continue to develop, children will have easier access to interdisciplinary expertise. Patients are most likely to be diagnosed and treated properly when managed by a specialist or team focused on these conditions. Recent insight into the etiopathogenesis of infantile hemangioma and vascular malformations may lead to novel therapies for these lesions in the near future.