2普萘洛尔处理婴幼儿血管瘤内皮细胞的分子机制

Propranolol treatment of infantile hemangioma endothelial cells： A molecular analysis. Stiles J, Amaya C, Pham R, et al. Exp Ther Med, 2012,4（4）：594-604.婴幼儿血管瘤（Hs）是一种好发于皮肤的良性血管性肿瘤,新生儿发病率约为5％～10％。     

2004'全国口腔颌面部脉管性疾病学术研讨会纪要

Congenital vascular anomalies have been the subject of much controversy and confusion over the years, many remain to be investigated and resolved. Authorized by the Managing Director Board of Chinese Stomatological Association (CSA), 2004 Chinese National Conference on Oral and Maxillofacial Vascular Anomalies was successfully held on July 15 to 18, 2004 in Shenzhen Grand Hotel. The conference was sponsored by Chinese Society of Oral and Maxillofacial Surgery (CSOMS), and undertaken by the Second People‘s Hospital of Shenzhen city. Professor ZHANG Zhen-kang, president of CSA, Professor QIU Wei-liu, president of CSOMS and academician of Chinese Academy of Engineers, and Professor LIU Bao-lin, vice president of CSOMS were present and made important speech at the meeting. More than 120 delegates nationwide attended this conference. Through a 3-day of oral presentation and discussion, the terminology, classification,treatment selection and outcome measurement of oral and maxillofacial hemangioma and vascular malformations were concerted among the delegates. Hemangiomas and vascular malformations have been recognized as distinct diseases that exhibit unique properties and behavior that demand an appropriately tailored treatment plan. The classification of Waner and Suen was adopted and different treatment modalities were reviewed and advocated for different lesions. Delegates were most interested in intralesional injection of Pingyangmycin for venous malformations, Krypton laser photodynamic therapy of venular malformations, Nd:YAG laser therapy for deep head and neck venous malformations after surgical exposure of the lesions, as well as “double“ embolization of large venous malformations reported by Professor QIN Zhong-ping. The conference witnessed the foundation of the Division of Vascular Anomalies, Chinese Society of Oral and Maxillofacial Surgery.     

国际文摘

1婴幼儿血管瘤和脉管畸形β1肾上腺素受体mRNA水平的差异β(1)-adrenoceptor mRNA level reveals distinctions between infantile hemangioma and vascular malformations.R觟ssler J,Haubold M,Gilsbach R,et al.Pediatr Res,2013Jan 31[Epub ahead of print]婴幼儿血管瘤(IH)是婴幼儿时期最常见的血管肿瘤。近年来,普萘洛尔,一种非选择性β1和β(2)肾     

国际文摘

1局部应用噻吗洛尔治疗小型婴幼儿血管瘤Topical timolol for small hemangiomas of infancy.MoehrleM,Léauté-Labrèze C,Schmidt V,et al.Pediatr Dermatol,2012Apr 4.[Epub ahead of print].普萘洛尔已经成为治疗巨大复杂婴幼儿血管瘤的首选药物,但是,关于其用药的安全性尚存在争议。该文作者介绍了局     

侵袭性牙周炎患者外周血CD-4+CD-25+调节性T细胞的检测及功能分析

Objective To investigate the changes of proportion and suppression function of CD-4+ CD-25+ regulatory T cells in the peripheral blood of patients with aggressive periodontitis.Methods Flow cytometric analysis was used to detect the frequency of CD-4+ CD-25+ regulatory T cells in the peripheral blood of 16 patients with generalized aggressive periodontitis and 17 patients with chronic periodontitis,as well as 17 periodontal healthy controls.Furthermore,CD-4+ CD-25+ regulatory T cells and CD-4+ CD-25- T cells were separated from peripheral blood of each enrolling subject using magnetic cell sorting technology.The direct suppression effect of CD-4+ CD-25+ regulatory T cells on CD-4+ CD-25- T lymphocytes proliferation was performed by the mixed lymphocytes reaction and measured by 3H-thymidine radioactive assay.Results The patients with generalized aggressive periodontitis had a lower frequency of CD-4+ CD-25+ regulatory T cells(9.71±4.01)%in the peripheral blood than periodontal healthy controls [(14.72±3.51)%]and chronic periodontitis patients[(17.01±5.16 )%],P＜0.05.A significant decrease was found in the suppression function of CD-4+ CD-25+ regulatory T cells from peripheral blood of patients with generalized aggressive periodontitis when co-cultured with CD-4+ CD-25- T lymphocytes in the proportion of 2:1,1:1 and 1:2 as compared with chronic periodontitis patients and periodontal healthy controls(P＜0.05).Conclusions Diminished numbers and decreased suppression function of CD-4+ CD-25+ regulatory T cells were found in patients with generalized aggressive periodontitis.     

侵袭性牙周炎患者外周血CD-4+CD-25+调节性T细胞的检测及功能分析

Objective To investigate the changes of proportion and suppression function of CD-4+ CD-25+ regulatory T cells in the peripheral blood of patients with aggressive periodontitis.Methods Flow cytometric analysis was used to detect the frequency of CD-4+ CD-25+ regulatory T cells in the peripheral blood of 16 patients with generalized aggressive periodontitis and 17 patients with chronic periodontitis,as well as 17 periodontal healthy controls.Furthermore,CD-4+ CD-25+ regulatory T cells and CD-4+ CD-25- T cells were separated from peripheral blood of each enrolling subject using magnetic cell sorting technology.The direct suppression effect of CD-4+ CD-25+ regulatory T cells on CD-4+ CD-25- T lymphocytes proliferation was performed by the mixed lymphocytes reaction and measured by 3H-thymidine radioactive assay.Results The patients with generalized aggressive periodontitis had a lower frequency of CD-4+ CD-25+ regulatory T cells(9.71±4.01)%in the peripheral blood than periodontal healthy controls [(14.72±3.51)%]and chronic periodontitis patients[(17.01±5.16 )%],P＜0.05.A significant decrease was found in the suppression function of CD-4+ CD-25+ regulatory T cells from peripheral blood of patients with generalized aggressive periodontitis when co-cultured with CD-4+ CD-25- T lymphocytes in the proportion of 2:1,1:1 and 1:2 as compared with chronic periodontitis patients and periodontal healthy controls(P＜0.05).Conclusions Diminished numbers and decreased suppression function of CD-4+ CD-25+ regulatory T cells were found in patients with generalized aggressive periodontitis.     

颏部形态与正畸矫治拔牙设计的相关性研究

Objective To determine the influence of chin on extraction in borderline cases.Methods Pre-treatment lateral cephalograms of 50 treated patients (25 first premolar extraction cases, 25 non-extraction cases) were collected and digitized. The average pre-treatment age was 16.5 years (15～18 years). 19 cephalometric parameters were used for assessment. Comparison was made between extraction and non-extraction orthodontic treatment in borderline cases. The correlation analysis, t-test and Logistic regression analysis were used. Results Significant hard tissue changes were found between extraction and non-extraction cases. Extraction cases had smaller chin prominence. Logistic regression analysis showed that the probability of orthodontic extraction was significantly correlated with Pog-NB. Conclusions The important factor in determining extraction in borderline cases was the prominence of the chin. The probability of orthodontic extraction decreased with the increasing chin prominence.     

牙齿移动过程中牙槽骨显微结构动态变化的微型CT研究

Objective To observe the microstructure changes of rat alveolar bone around tooth root under orthodontic force loading. Methods Ten 10-week-old rats were included in the study. Upper first molars were moved mesially with 0.196 N of force. The alveolar bone around the root of upper first molar was scanned by a micro-computed tomography scan system(SkyScan-1076) in different period after the initiation of orthodontic force loading( on the 3rd, 7th, 14th day after force loading ) and analyzed by a speciallydesigned software to measure the microstructure parameters of alveolar bone ( bone volume fraction, bone surface to volume ratio, structure model index, trabecular thickness, trabecular separation, trabecular number). Results From the 7th day, bone volume fraction[(41±14)%], structure model index( 1.51 ±0. 52) and trabecular separation [(90 ± 30 ) μm] changed significantly in the compressive area compared with those[(64±15)%, (0.51 ±0.85), (56 ±-10) μm] on the 3rd day. From the 14th day, bone volume fraction[(78 ± 14)%], structure model index(0. 28 ± 0. 20) and trabecular separation[(29 ±13) μm] changed significantly in the tension area compared with those[(67 ± 14)%, (0.40 ±0.41 ),(48 ± 15) μm] on the 7th day. No difference on trabecular number was found between the compressive area and tension area(P ＞0. 05). Conclusions The significant resorption of the alveolar bone was observed on the 7th days in the compressive area and the deposition of the alveolar bone was observed on the 14th day in the compressive area after orthodontic force loading.     

Diagnostic template research of craniofacial characters of Class Ⅱ patients

Objective To analyze the morphological characters and mechanism of Class Ⅱpatients. Methods 894 Class Ⅱ patients were collected from the orthodontic department, Peking University School of Stomatology from 1997 to 2000. The standardized coordinates of thecephalometric radiograms were obtained by Procrusts Superimposition (PS). Cluster analysis was carried out to divide the patients into different subdivisions and the cephalometric character of each template was analyzed. Resulst 894 patients were divided into 11 subdivisions with specialcharacteristics. The morphological templates were constructed by computer. Conclusions Procrusts method was effective to study the classification of the malocclusion. According to the template and its cephalometric value, Class Ⅱ malocclusion were mainly due to the mandiblar retrusion, and verticaldiscrepancy present in 60percent of the patients.     

锥形束CT转化头颅侧位片定点精确性的研究

Objective In this study, we compared the precision of landmark identification using cephalograms from cone-beam computed tomographic (CBCT) volumes and conventional lateral cephalograms (Ceph). Methods Twenty pre-orthodontic patients were radiographed with conventional Ceph and CBCT techniques. CBCT data was uploaded into InvivoDental software 5.0 to generate cephalograms(CCB). Three observers plotted 23 landmarks using computer displays of CCB and Ceph views during separate sessions. Absolute differences between CCB and Ceph of all observers were measured. The absolute difference between each observer was also measured. ANOVA and paired t tests were used to analyze variability differences. Results The difference of landmark identification between CCB and Ceph were significant at P＜0.05 calculations. 8 landmarks variability was statistically greater than Ceph views. The variability of CCB for each observer was consistently greater than in Ceph. The overall correlation of CCB and Ceph measurements was excellent at 0.99.Conclusions The CCB displays of CBCT volume images provide generally more precise identification than Ceph. More precise location of basion, porion, orbitale, ANS, A point, Gonion and tooth apex overcomes the problem of superimposition of these bilateral landmarks seen in Ceph. Greater variability of certain landmarks is probably related to inadequate definition of the landmarks in the third dimension.     

血管内皮细胞体外培养及生物学特性的探讨

目的　分离培养脐静脉内皮细胞 ,鉴定其生物学特性。方法　采用 0 2 %胰蛋白酶行脐静脉两次酶灌注消化法获取内皮细胞。分别在倒置相差显微镜、透射电镜下观察 ,SP免疫组化法行Ⅷ因子抗原鉴定。观察血管内皮细胞体外培养的生长情况 ,并绘制细胞生长曲线。结果　 (1)经改良的胰蛋白酶两次消化法获取较高的活细胞百分数 ,且细胞纯度较高。 (2 )透射电镜下发现多数细胞间存在紧密连接结构 ,胞膜的一侧贴有基膜样成分。 (3)血管内皮细胞体外培养融合成单层后 ,形成类似体外血管内皮的内衬结构。结论　透射电镜下细胞特征可作为鉴定血管内皮细胞的辅助手段。     

16型人乳头状瘤病毒E6、E7诱导的人永生化口腔上皮细胞系的建立

目的：建立16型人乳头状瘤病毒（human papiloavirus,HPV16)E6,E7诱导的永生化口腔上皮细胞系，方法：正常口腔上皮细胞转染pLXSNHPV16E6,E7,G418筛选阳性细胞，连续培养建系；Western blot检测细胞HPV16E6，E7蛋白表达；生物学特性检测。集落实验；成瘤实验，结果：细胞连续培养达18个月余，命名为永生化口腔上皮细胞（human immortalized oral epithelial cell,HIOEC) ,HIOEC表达HPV16E6，E7蛋白，角蛋白阳性，透射电镜观察细胞有丰富的张力原纤维，细胞之间形成桥粒，克隆形成率0．77％，裸鼠未成瘤。结论：成功建立了HPV16E6，E7诱导的人永生化口腔上皮细胞系。     

Establishment of mesenchymal cell line derived from human developing odontoma

Oral Diseases (2012) 18, 756-762 Objectives: An odontoma, which shows proliferating odontogenic epithelium and mesenchymal tissue, is one of the most common odontogenic tumors encountered. These are commonly found in tooth-bearing regions, although the etiology remains unknown. There are no previous reports of an established line of immortalized human odontoma cells. Methods: Using odontoma fragments obtained from a girl treated at our department, we established an immortalized human odontoma cell line and investigated cell morphology, dynamic proliferation, the presence of contamination, and karyotype. Moreover, cell characterization was examined using osteogenic and odontogenic markers. Results: We successfully established a mesenchymal odontoma cell (mOd cells). The cells were found to be fibroblastic and had a high level of telomerase activity. Cell growth was confirmed after more than 200 population doublings without significant growth retardation. mOd cells expressed mRNA for differentiation markers, including collagen type I (COLI), alkaline phosphatase, bone sialoprotein, osteopontin, osteocalcin, cementum-derived protein (CP-23), dentin sialophosphoprotein (DSPP), and distal-less homeobox 3 (DLX3), as well as bone morphogenetic proteins (BMPs). In addition, they showed a high level of calcified nodule formation activity in vitro. Conclusions: We successfully established a cell line that may be useful for investigating the mechanisms of normal odontogenesis as well as characteristics of odontoma tumors.     

慢病毒介导过表达端粒酶逆转录酶的 人口腔黏膜上皮稳定细胞系的构建

目的 通过慢病毒法建立稳定表达外源性人端粒酶逆转录酶（hTERT）基因的口腔黏膜上皮细胞（OMECs）,探索构建高效、稳定的永生化OMECs细胞系的方法。方法 提取293T细胞总RNA,应用聚合酶链反应（PCR）法扩 增hTERT基因全长,构建重组慢病毒载体pLVX-puro-hTERT。包装慢病毒颗粒后感染人正常OMECs,经嘌呤霉素抗 性筛选获得阳性克隆,采用实时荧光定量PCR法和Western blot法检测hTERT基因mRNA和蛋白的表达水平。结果 成功构建了pLVX-puro-hTERT过表达慢病毒载体并感染到OMECs中;感染细胞与正常OMECs形态相似,呈铺路石 样生长;实时荧光定量PCR和Western blot结果均显示,hTERT在感染细胞中高表达,与正常细胞相比差异有统计学 意义（P<0.05）。结论 通过慢病毒法成功建立了过表达hTERT的OMECs稳定细胞系,为构建高效、稳定增殖的人 永生化OMECs细胞系奠定了实验基础。     

免疫脂质体鱼肝油酸钠作用于人血管瘤内皮细胞的实验研究

目的：探讨免疫脂质体鱼肝油酸钠的制备方法及其对人增殖期血管瘤内皮细胞的作用。方法：以N-琥珀酰-3-2-联硫基吡啶-丙酸盐（（N-Succinimidyl-3-（2-pyridyldithio））propionate,SPDP）作为耦联剂,将针对血管内皮生长因子受体2（VEGFR2/KDR）的单克隆抗体耦联到脂质体表面,运用吹膜挤压法制备免疫脂质体鱼肝油酸钠,通过激光共聚焦显微镜和倒置显微镜观察、Giemsa染色、透射电镜观察、MTT以及流式细胞仪检测,探讨其对人增殖期血管瘤内皮细胞的作用。结果：制备的免疫脂质体鱼肝油酸钠平均粒径为122.9nm,具有良好的稳定性;与普通脂质体制剂相比,其与人血管瘤内皮细胞结合能力更强、更早,促凋亡作用更为明显,而对人血管瘤内皮细胞的毒性作用与普通脂质体制剂类似,作用较为缓和。结论：成功制备免疫脂质体鱼肝油酸钠,其在体外具有良好的特异性主动靶向能力,可以诱导人增殖期血管瘤内皮细胞发生较为普遍的凋亡。     

裸鼠皮下种植性人头颈鳞癌模型的建立

目的：建立稳定的皮下种植性裸鼠人头颈鳞癌模型。方法：取10^9个对数期的PCI-37B细胞,种植于BALB／c裸鼠皮下。待成瘤后,从裸鼠体重、大体形态,肿瘤生长情况,病理学特征等方面进行研究。采用SPSS13．0软件包对数据进行统计学分析。结果：肿瘤形成的潜伏期约15d,实验组14只裸鼠成瘤13只,肿瘤成功率为92．8％。种植肿瘤组裸鼠体重较正常对照裸鼠明显减轻．肿瘤生长迅速．病理学检查有明显的细胞异形性。结论：成功建立了裸鼠皮下种植性头颈鳞癌模型。本模型具有在短期内易建立、稳定、重复性好、成癌率高等特点,为人头颈鳞癌的生物学、治疗学研究提供了良好的动物模型。     

Isolation and characterization of an immortalized oral keratinocyte cell line of mouse origin

OBJECTIVE: To establish an oral epithelial cell line of mouse origin for molecular and biochemical assays. DESIGN: Epithelial cells were isolated from the oral cavity of adult mice and established as a spontaneously immortalized cell line in culture, designated immortalized oral keratinocyte cells (IMOK cells). The cells were then characterized for growth characteristics, differentiation potential, karyotype, transfectability, susceptibility to viral infection and responses to siRNA. RESULTS: The IMOK cells exhibit robust growth in both serum-containing and serum-free medium for at least 100 population doublings. IMOK cells have a near diploid karyotype, express keratinocyte marker proteins and can be induced to undergo differentiation by the addition of high levels of calcium to the medium. The differentiation process is characterized by morphological changes and by the induction of oral epithelium specific differentiation marker proteins such as K4 and K13. Transient transfection analyses reveal that IMOK cells are highly transfectable and that several promoters of epithelial cells are active in these cells. Moreover, upon differentiation with calcium, there is an up-regulation of differentiation-specific K4 and Elf5 promoter activity. Finally, we show that the oral keratinocytes are also amenable to infection with retroviruses and to siRNA-based knockdown of gene expression. CONCLUSIONS: Our study is the first to establish an immortalized oral keratinocyte cell line of murine origin that can recapitulate the oral epithelium differentiation program and thus could serve as a useful tool for toxicological and molecular analyses of the oral tissue.     

复发性涎腺腺样囊性癌裸鼠移植瘤模型的建立及其生物学特性

目的建立人复发性涎腺腺样囊性癌裸鼠移植瘤动物模型。方法将人复发性涎腺腺样囊性癌组织块,进行裸鼠皮下接种、传代,建立裸鼠移植瘤模型,并对其进行光镜、电镜、免疫组织化学及染色体分析。结果人复发性腺样囊性癌组织块移植于裸鼠后,获原代移植成功。2年期间移植瘤在裸鼠之间共连续传代4代,移植BALB/C裸鼠5只,平均潜伏期44.33天。经光镜、电镜、免疫组织化学观察发现移植瘤与其原发肿瘤结构特征一致。染色体检查为亚中着丝点染色体,符合人类体细胞核型。结论成功建立人复发性涎腺腺样囊性癌裸鼠移植瘤模型并传代,移植瘤保留了原发肿瘤的组织学特征和染色体核型。     

组织块法建立婴幼儿血管瘤动物模型的实验研究

目的 ：探索建立婴幼儿血管瘤动物模型的方法。方法 ：将16只裸鼠随机分为2组,一组每周肌注雌二醇1次,每次0.1 mg;另一组为对照组,将不同类型的婴儿颌面部血管瘤标本分别植入16只裸鼠体内,观察其生长状况,测量其体积变化,并于90 d时进行病理学检查,进行CD31、CD34、Ki67免疫组织化学染色和CD34免疫荧光检测。应用SPSS13.0软件包对数据进行统计学分析。结果 ：实验组移植瘤体显著大于对照组（P<0.05）,其生长特点与人血管瘤相似。光镜下检测符合血管瘤增殖期的病理特点,CD31、CD34、Ki67检测呈强阳性,与裸鼠瘤周正常肌肉组织相比,差异显著（P<0.05）。结论 ：通过组织块移植法建立婴幼儿血管瘤模型是一种可靠的方法,为婴幼儿血管瘤的研究提供了平台。