PURPOSETo observe the effect of gene therapy mediated by oral administration of attenuated Salmonella carrying tip30 and IFN-γ genes in human tongue carcinoma nude mouse model.METHODS25 four-week-old BALB/C male nude mice were divided randomly into 5 groups based on the differently harboring genes in the recombinant attenuated Salmonella typhimuriums SL7207, which included blank group (PBS), control group (SL7207), IFN treatments group (SL7207-pCI-IFN), tip30 treatment group (SL7207-pCI-tip30) and the combination of IFN and tip30 treatment group (SL7207-pCI-tip30/IFN). On 10d after submandibular subcutaneous injection of Tca8113 cells into the mice, the recombinant attenuated Salmonella typhimuriums were orally administrated 3 times at 1 week interval. The treatment effect indicators included the growth curve, the tumor inhabitation rate, the survival rate, the apoptosis typical DNA ladder and the protein expressions of tip30 and IFN-γ in tumor cells. Statistical analysis was performed with SPSS15.0 software package.RESULTSThe growth curve, the tumor inhabitation rate and the survival rate indicated that the tumor in the PBS group constantly grew up and induced the animal death in earlier time, while the SL7207 treatment had a slight inhibiting effect compared with PBS group. However, the SL7207-pCI-IFN group and the SL7207-pCI-tip30 had a moderate inhibiting effect compared with the SL7207 group, while the combination of IFN and tip30 had the strongest inhibiting effect. The protein expressions of tip30 and IFN-γ were detected in tumor cells while the typical DNA ladders of apoptosis were observed only in the tip30 gene transfer groups (SL7207-pCI-tip30 and SL7207-pCI-tip30/IFN).CONCLUSIONSThe recombinant attenuated Salmonella typhimurium possesses a capacity of gene transfer in vivo through oral administration. The combination of the harboring tip30 and IFN-γ genes have synergistic inhibiting tumor effect for tongue carcinoma.

目的 观察重组减毒鼠伤寒沙门菌作为基因载体在人涎腺腺样囊性癌裸鼠移植瘤中的富集情况和对外源基因的呈递能力.方法 以绿色荧光蛋白基因(GFP)为报告基因,以减毒鼠伤寒沙门菌SL7207为转基因载体,分别构建原核启动GFP表达的重组减毒鼠伤寒沙门菌SL7207-pUC-GFP和真核启动GFP表达的重组减毒鼠伤寒沙门菌SL7207-pEGFPN1.原核菌SL7207-pUC-GFP在体外连续传代,观察GFP基因表达的稳定性；同时,对荷人涎腺腺样囊性癌皮下移植瘤裸鼠模型口服给予原核菌SL7207-pUC-GFP(0.1 mL,1×109cfu/mL),在口服后24 h、48 h、5d、10d、20 d、30 d处死荷瘤裸鼠,获取肝、脾及肿瘤组织并制成匀浆液进行重组菌培养及GFP表达检测,观察重组菌在瘤体细胞内富集情况.对荷瘤裸鼠模型口服给予真核菌SL7207-pEGFP-N1,5d后取肝、脾及肿瘤组织进行冰冻组织切片,荧光显微镜下观察GFP的表达,了解重组菌携带外源基因在肿瘤细胞内的表达.结果 携带GFP原核表达的重组减毒鼠伤寒沙门菌SL7207-pUC-GFP在体外连续传代10次未见表达减少或缺失.荷瘤裸鼠口服原核表达GFP基因重组菌SL7207-pUC-GFP菌液实验表明,重组菌SL7207-pUC-GFP在肝、脾及肿瘤组织中能长期存活,以肿瘤组织中聚集明显(P＜0.05)且维持时间较长.荷瘤裸鼠口服真核表达GFP基因重组菌SL7207-pEGFP-N1菌液实验表明,相对肝脏和脾脏组织,外源基因在肿瘤细胞内表达量最高.结论 重组减毒鼠伤寒沙门菌可以在瘤体细胞内富集存活,并且携带的外源基因可以释放到肿瘤细胞内表达,具有作为基因治疗载体的双重优势.

PURPOSE:To explore the effectiveness of Osteopontin siRNA(OPNsiRNA) on viability and invasiveness of Tca 8113 by attenuated Salmonella typhimurium.METHODS: Synthesised OPNsiRNA and nonsense ControlsiRNA were separately constructed into expression vector pIRES2-EGFP,and were transferred into attenuated Salmonella SL7207.The tongue cancer cell line Tca 8113 was infected by the recombinant Salmonella SL7207,named SL-OPNsiRNA and SL-ControlsiRNA respectively.The infection rate,growth,apoptosis,migration and invasion were observed while the protein expressions of OPN,MMP-2 and uPA were detected by Western blot.SPSS15.0 software package was used for statistical analysis.RESULTS: The infection rates of the recombinant Salmonellas both were more than 80.0% without significant difference(P>0.05).The growth was significantly inhibited and the apoptosis was significantly induced after infection of SL-OPNsiRNA compared with SL-ControlsiRNA.After 24h-infection of SL-OPNsiRNA and SL-ControlsiRNA,the migration index was 0.56±0.10 and 0.82±0.05(n=12,P<0.01),the invasion index were 0.53±0.13and 0.83±0.06(n=12,P<0.01),respectively,and the intra-cell proteins of OPN,MMP-2 and uPA were also significantly down-regulated by OPNsiRNA compared with ControlsiRNA.CONCLUSIONS:OPNsiRNA could be effectively transferred into tongue cancer cells through attenuated Salmonella,and inhibited caner cell growth and metastasis.

PURPOSE To culture primary mouse odontoblast and to provide a base for study on inducing ES cells to odontoblast. METHODS Lower incisor germs were removed from 1-week-old mouse. The dental papillae were isolated in microscope, and the dental papillae cells were dispersed using 0.25% collagenase and 0.25% trypsin. The cell clones, which had similar morphology to odontoblasts, were selected for further culturing. The primary cultured cells were identified by light and electron microscopes and mRNA expression of mouse dentin sialophoprotein. RESULTS The cultured cells had the same morphology and ultrastructure. They were rich in Golgi's complex, ribosome and rough endoplasmic reticulum. These cells expressed DSPP at mRNA levels. CONCLUSION The cultured cells were mouse odontoblast-like cells. The method could be used for the study of odontal cells in vitro.

To evaluate the clinical results of ceramic optimized polymer (Targis) posterior inlays.345 type I cavities were divided into two groups: 170 in Targis inlays group (42 premolars and 128 molars) and 175 in control group (composite resin inlays, 45 premolars and 130 molars). The clinical effects of Targis inlays and composite resin inlays in posterior teeth were compared in marginal discoloration, marginal adaption, secondary caries, anatomic shape integrity, fractures of tooth and color match 3 years later. The data were analyzed using Chi-square test.3 years after treatment, 166 Targis inlays were followed up, 2 had marginal discoloration, 158 had good marginal adaption, 162 had anatomic shape integrity, 1 had secondary caries, 161 had color match, and 5 had fractures of teeth. 169 composite resin inlays were followed up, 27 had marginal discoloration, 134 had good marginal adaption, 150 had anatomic shape integrity, 20 had secondary caries, 164 had color match,5 had fractures of teeth. The marginal discoloration, marginal adaption, secondary caries, anatomic shape integrity of Targis inlays were better than that of composite resin inlays (P < 0.05), but fractures of teeth was not different between inlays in the two group (P > 0.05).Targis posterior inlays is a good newly developed prosthesis.

PURPOSE:To investigate the methods of induction of mouse embryonic stem cells to differentiate into odontoblast-like cells by co-culture with pulp fibroblast.METHODS:By suspension culture, embryonic stem cells were induced to form embryoid bodies. Then the cells from embryoid bodies were co-cultured with pulp fibroblast in Transwell system for differentiation toward odontoblast-like cells.RESULTS:By RT-PCR analysis, embryoid body cells gave rise to the cell population expressing DSPP, a specific odontoblast cell marker, after 10 days of co-culture with pulp fibroblast and the expression of DSPP was enhanced after 15 days of co-culture. On the other hand, the expression of DSPP was not detected in the isolately cultured embryoid body cells.CONCLUSIONS:Embryonic stem cells can be induced into odontoblast-like cells by co-culture with pulp fibroblast. Supported by Science and Technology Development Fund of Shanghai Municipality (Grant No. 03ZR14027).

PURPOSE:To evaluate the clinical effect of two light cured pit and fissure sealants for preventing pit and fissure caries.METHODS:416 health permanent teeth in 162 children aged 6 to 8 years old children were divided into two groups, enameloplasty sealant technique (EST) and cup-shaped brush sealant technique (CST) were used in two groups, respeitively. 90 and 112 contralateral healthy teeth were used as self controls respectively. The rate of the sealant reservation and the incidence of caries were compared in the two groups, and the clinical effect of the two groups was compared using SPSS10.0 software package for Chi-square test.RESULTS:After 3 years, the rate of the sealant reservation and the incidence of caries of EST were 85.4% and 0.9% respectively, the rate of the sealant reservation and the incidence of caries of CST were 63.8% and 6.7% respectively. Statistical analysis revealed a significant difference (P<0.05) between the two groups.CONCLUSION:EST and CST both have good clinical effects, While EST is superior to CST in preventing pit and fissure caries.

Objective:To observe the inhibitory effect of recombinant attenuated Salmonella typhimurium(AST) harboring eukaryotic expression plasmids encoding (Tip30, IFN-g and Tip30 fused with human IFN-g gene) on tumor angiogenesis in BALB/c mice with human salivary adenoid cystic carcinoma. Methods: Twenty-five 4-week-old BALB/c male nude mice were equally randomized into 5 groups: control group (PBS group), simple attenuated Salmonella group (SL7207 group), AST harboring plasmid pCI-IFN group (SL7207/pCI-IFN group), AST harboring plasmid pCI-Tip30 group(SL7207/pCI-Tip30 group), and AST harboring co-expressing plasmid pCI-Tip30/IFN group (SL7207/pCI-Tip30/IFN group). BALB/c mice in all groups received submandibular administration of ACC-2 cells suspension. The control group received oral PBS 10 d after injection, and the other groups received different recombinant Salmonella typhimuria 3 times weekly. Then the tumor size, tumor-bearing survival time and tumor microvascular density (MVD) in ACC-2 tumor were determined. Results: Compared with the control group, the tumor sizes of the 4 treatment groups were smaller and the tumor-bearing survival times were longer (P 0.05). Compared with the SL7207 group, tumor sizes of Tip30 and human IFN-g gene treatment groups were smaller and the tumor-bearing survival times were longer (P0.05). Compared with the single gene groups, the tumor size of the fusion genes treatment group were smaller and the tumor-bearing survival time were longer (P0.05). Compared with the other 3 groups, the microvascular of density the 2 treatment groups harboring Tip30 gene was lower. Compared with the treatment group harboring Tip30 gene, the microvascular density of the fusion gene treatment group was lower (P0.05). Conclusion: Both AST-mediated Tip30 gene treatment and IFN-g gene treatment can inhibit tumor growth, and the fusion gene treatment of the 2 has better inhibitory effect than the single gene, indicating that there is synergy between the 2 genes.

Objective: To observe inhibitor of 4 kinds constructed attenuated Salmonella typhimurium on the lungs metastasis transplant tumor growth in BALB/C nude mice with human salivary adenoid cystic carcinoma. Methods: 25 four-week-old BALB/C male nude mice were divided randomly into 5 groups, each group had 5 mice. They were the control group (PBS), the simple attenuated Salmonella group (SL7207), the recombinant attenuated Salmonella typhimuriums haboring plasmid pCI-IFN therapy group(SL7207/pCI-IFN), the recombinant attenuated Salmonella typhimuriums haboring plasmid pCI-Tip30 therapy group (SL7207/pCI-Tip30) and the recombinant attenuated Salmonella typhimuriums haboring co-expressing plasmid pCI-Tip30/IFN therapy group(SL7207/pCI-Tip30/IFN). All groups were injected ACC-M cells suspension at the tail vein. The control group accepted PBS by oral administration 10 days after injection, and the other groups accepted the different recombinant Salmonella typhimuriums respectively, once every 2 weeks and 3 times in all. Then the ACC-M lung metastasis tumor were detected with the expressions of Tip30 and human IFN-γ gene, tumor size, tumor weight, and mice survival period with tumor. Results: There were Tip30 and human IFN-γ gene expression in cytoplasm of ACC-M lungs metastasis transplant tumor cells in all therapy groups. The tumor size and weight of the therapy groups were lower than that of the control group, the rate of suppressing of the therapy groups were higher than that of the control group and mice survival period of therapy groups were longer than the control group. In the therapy groups haboring Tip30 and human IFN-γ gene, the tumor size, tumor weight were lower than that of the SL7207 group, the rate of suppressing were higher than that of SL7207 group and mice survival period were longer than that of the SL7207 group. In the united genes therapy group, the tumor size, tumor weight were lower than that of the single gene groups, the ratio of suppressing tumor were higher than that of the single gene group and mice survival period were longer than that of the single gene groups. Conclusion: Both the attenuated Salmonella typhimurium and Salmonella typhimurium had evident inhibitor effect on the lungs metastasis transplant tumor growth in BALB/C nude mice with human salivary adenoid cystic carcinoma. The united genes may have better inhibitor effect than single gene, because there are synergism between them.

Objective: To investigate the object analysis of lymph node pathologic lesion by using magnetic resonance imaging. Methods: For 22 patients suffering various kinds of disorders involving lymph nodes in the head and neck, magnetic resonance imaging was applied in assessment of lymph nodes status and compared with histological sections. Results: In 22 cases, the coincidence rate of MRI with the pathologic diagnosis was 95%(21/22). Conclusion: MRI could provide objective evidence for diagnosis of lymph node in the head and neck, which was one of the important examinations in detecting various kinds of disorders involving lymph node in the head and neck.

涎腺肿块可以是良性肿瘤或恶性肿瘤,也可是囊肿或炎性肿块,既可来自腺体,也可来自腺外组织,给诊断造成了困难.现总结上海长海医院口腔科从1998年3月～2003年3月间,收入病房行手术治疗的521例患者的临床资料,报告如下.

本实验利用减毒沙门菌的肿瘤靶向性,以其为载体,携带抑癌基因Tip30和人IFN-γ基因,通过口服方式对舌癌进行基因治疗动物实验,以期在临床上为舌癌的基因治疗提供动物实验依据.

AIM:To provide theoretical basis and experimental evidence for diagnosis and therapy of indirect injuries of dental pulp in gunshot wounds.METHODS:The levels of IL-1 in different stages were studied using immunohistochemical method to evaluate its functions in inflammatory process of dental pulps.RESULTS:IL-1 expression was detected by immunohistochemical assay in postwound conditions.The results showed that positive staining of IL-1 was observed predominantly on infiltrating inflammatory cells,vasculars endothelial cells and some fibroblasts,which was consistent to the recruitment and activation of inflammatory cells,and with the severity duration of pulpal tissue injury and response.CONCLUSION:These results indicate that the IL-1 is involved in regulating physiologic responses of dental pulp injuries in maxillofacial trauma by a projectile.

The ameloblasts are the key cells during odontogenesis,a nd they have been already cultured in vitro . The amelogenin is the extracellul ar matrix synthesized and secreted by the ameloblasts, and it plays a crucial role during amelogenesis. The multiplication and differentiation of the ameloblasts w ere controlled by many factors. This article reviews the culture of ameloblasts, the study of amelogenin and the controlling factors of the multiplication and d ifferentiation of the ameloblasts.

Objective: To construct attenuated salmonella typhimurium haboring an eukaryotic co-expression plasmid encoding TIP30 and human IFN-γ gene and observing its effect on the growth of adenoid cystic carcinoma. Methods: The TIP30 and human IFN-γ genes were amplified by PCR and inserted into eukaryotic expression vector pCI-neofor the construction of expression plasmids pCI-TIP30 and pCI-IFN, respectively. A co-expression plasmid pCI-TIP30/IFN was constructed by linking TIP30 and human IFN-γ gene using the sequence of internal ribosome binding sequence (IRES). Three recombinant expression plasmids were transformed into an attenuated AroA'autotrophic mutant of salmonella typhimurium SL7207, the resultant bacteria were used to infected murine macrophage in vitro and the expressed products were detected by Western blot and ELISA, respectively. Tumor growth was observed by oral administration of the recombinant salmonella typhimuriums to the nude mouse with adenoid cystic carcinoma. Results: Murine macrophage infected with recombinant salmonella transformed with both plasmids pCI-TIP30 and pCI-TIP30/IFN could express TIP30 protein, and murine macrophage infected with recombinant salmonella transformed with pCI-IFN or pCI-TIP30/IFN could secret human IFN-γ in the culture supernatant. Attenuated salmonella typhimurium and three constructed recombinant salmonella typhimuriums all had evident inhibition onthe tumor growth in nude mouse with adenoid cystic carcinoma. Conclusion: The recombinant attenuated salmonella typhimuriums haboring plasmid pCI-TIP30, plasmidpCI-IFN and co-expressing plasmidp-CI-TIP30/IFN were successfully constructed, which could inhibit the growth of adenoid cystic carcinoma in nude mouse.

目的:研究白细胞介素-8（IL-8）在狗颌面部火器伤间接损伤牙髓中的表达情况。方法:采用直径5.80mm制式枪弹致伤狗下颌部的模型,并利用免疫组化染色及定量分析方法检测颌面部火器伤后不同时间、下颌不同牙齿的牙髓组织中IL-8的动态变化。结果:IL-8在伤后牙髓组织中有阳性表达,并随时间的改变,呈动态变化过程。结论:狗牙髓的血管内皮细胞能分泌合成IL-8,可能参与火器性牙髓炎症发生、发展,并与牙髓损伤、修复有关。

自1998年人类多能干细胞首次被成功分离以来,干细胞的研究就一直受到公众的关注,一方面是由于它极为广阔的发展前景,另一方面则是由于它的合法性和伦理方面的争议.但国内外在口腔医学领域中研究才刚刚起步,本文对目前干细胞研究及在口腔医学领域中的进展作一综述.

目的比较替硝唑碘仿根管糊剂和甲硝唑碘仿根管糊剂在治疗慢性化脓性根尖周炎伴异位窦道疗效上的差异。方法选择慢性化脓性根尖周炎伴异位窦道的患者127例,随机分为2组,替硝唑碘仿根管糊剂组（实验组）和甲硝唑碘仿根管糊剂（对照组）。应用两次法与牙胶尖充填根管,记录临床症状和体征,6月后拍摄X线片,观察根尖阴影是否消失。结果实验组临床疗效优于对照组,且有统计学差异（P＜0.05）。结论替硝唑碘仿根管糊剂在治疗慢性化脓性根尖周炎伴异位窦道的疗效优于甲硝唑碘仿根管糊剂。

Objective To analyze the epidemiological characters of the original mass in the carotid triangle area. Methods This article had analyzed 267 cases of the original mass in the carotid triangle area. Results The incidence rate of the original benign tumor was 49. 06% , among which the branchial cleft cyst was the first rank, accounted for 23. 97% ; the vascular tumor was the second rank, accounted for 15. 73% . The incidence rate of the inflammation was 30. 71% , among which chronic lymph node reactive hyperplasia was the first rank, accounted for 18. 73% ; the tuberculous lymphadenitis was the second rank, accounted for 10. 01% . The incidence rate of the original malignant tumor was 20. 22% , among which the metastasizing malignant tumor was the first rank, accounted for 10. 49% . Conclusion The incidence rate of the original benign tumor in the original mass in the carotid triangle area was the first rank, the inflammation was the second rank, the malignant tumor was the third rank, the ratio was about 5:3:2.

髁状突骨折在下颌骨骨折中占有很高的比例,其发生率约为25%.它对牙颌功能以及儿童的面部生长发育影响最大,尤其是存在着发生真性颞下颌关节强直、颞下颌关节紊乱综合征的潜在危险,因此日益受到重视[1].我们在近5年中对35例髁状突骨折患者采取了保守治疗,取得了良好的临床疗效,现作一分析.