OBJECTIVETo evaluate the morphological characteristics of alveolar bone defects of the patients with chronic periodontitis using cone-beam CT (CBCT).

METHODSSixty patients with chronic periodontitis were included in this study. CBCT was used to scan the alveolar bone and NNT software to measure the alveolar bone defects and bone loss types in different regions.

RESULTSSeventy-five percent (45/60) of the alveolar bone defect was the generalized type, 25% (15/60) was the localized type. In incisor and canine area, the defect of the mandibular alveolar bone was more severe than in the same sites of maxilla. There was less bone loss in the premolar area of mandible than in the same site of maxilla. In the mesial and buccal sites of mandibular molars and in the lingual site of maxillary molars, the most severe alveolar bone loss was found.

CONCLUSIONSThe obvious alveolar bone defect areas in chronic periodontitis were the palatal side of maxillary molars and the lingual side of mandibular incisors. CBCT can clearly demonstrate the degree of alveolar bone defects in different regions of chronic periodontitis.

Adult ; Alveolar Bone Loss ; diagnostic imaging ; Chronic Periodontitis ; diagnostic imaging ; Cone-Beam Computed Tomography ; Female ; Humans ; Male ; Middle Aged

Objective: To investigate the preparation of bioactive denatured acellular dermal matrix (DADM) from burn mice riched in mice bone marrow mesenchymal stem cells. Methods: Twelve BALB/c mice were collected and 20% total body surface area scalds (hereinafter referred to as burns) with deep partial thickness were inflicted on the back skin of each mouse. After removing epidermis, the burned skin were collected and divided into Triton X-100 group and elhylene diamine tetraacetic acid (EDTA) group according to the random number table, with 15 samples in each group. Samples in Triton X-100 group and EDTA group were respectively placed in mixture of 2.5 g/L Triton X-100 and 2.5 g/L trypsin solution and mixture of 0.2 g/L EDTA and 2.5 g/L trypsin solution for sustained vibration and elution for 24 hours to make mice DADM. The general appearance of DADM was observed. The structure and arrangement of collagen fibers of DADM were observed by scanning electron microscope and tissue structure of DADM were observed by fluorescence microscope. Bone marrow mesenchymal stem cells (BMSCs) from mice were transplanted in mice DADM in the two groups with concentration of 2×105 cells per well to prepare bioactive mice DADM. After cultured for 3 days, tissue structure of bioactive mice DADM was observed by hematoxylin and eosin staining, distribution and number of BMSCs of bioactive mice DADM were observed by immunofluorescence staining. Proliferation of BMSCs of bioactive mice DADM after cultured for 2 h, 1 d, 3 d, and 5 d was detected by cell count kit-8. Data were processed with analysis of variance for repeated measurement and t test. Results: (1) Mice DADM in the two groups were white in appearance with certain tenacity and elasticity. Mice DADM in the two groups maintained good three-dimensional porous network structure. Collagen fibers of mice DADM in EDTA group were with good continuity, and collagen fibers of mice DADM in Triton X-100 group were fractured in varying degrees. Mice DADM in the two groups were decellularized completely, and the collagen fibers were loose and arranged disorderly. The continuity of tissue structure of mice DADM in EDTA group was better than that of mice DADM in Triton X-100 group. (2) After cultured for 3 days, the BMSCs in bioactive mice DADM in the two groups were evenly distributed. The number of bioactive BMSCs in mice DADM in EDTA group was 37±7, which was significantly more than that of mice DADM in Triton X-100 group (25±8, t=0.128, P<0.05). The proliferation of bioactive BMSCs in mice DADM in Triton X-100 group and EDTA group was similar at 2 hours and on day 1 after cultured (t=1.292, 0.656, P>0.05). On 3, 5 days after cultured, the proliferation of bioactive BMSCs in mice DADM in EDTA group was significantly higher than that of mice DADM in Triton X-100 group (t=2.309, 14.128, P<0.05 or P<0.01). Conclusions Mice DADM prepared by decellularization of EDTA has better three-dimensional porous network structure and good continuity of collagen fiber. The BMSCs in bioactive DADM from burn mice prepared by transplanting BMSCs are evenly distributed with large quantity and strong proliferative capacity, which has the potential to be good autologous dermal substitute.

The mutant population of Xanthomonas oryzae pv oryzae strain differential to rice bacterial blight resistance gene Xa23 has been constructed mediated by transposon in vivo . The results of PCR amplification with specific primers and analysis of flanking sequence of mutants indicated that the foreign DNA has been integrated into X. oryzae pv oryzae genome. Four mutants with changed avirulent activity to Xa23 gene have been identified by artificial inoculation. It is possible to clone genes that are required for AvrXa23 avirulence activity using this new strategy.
Bacterial Proteins ; genetics ; Base Sequence ; DNA Transposable Elements ; Gene Expression Regulation, Plant ; Genes, Plant ; Molecular Sequence Data ; Mutation ; Oryza ; genetics ; microbiology ; Plant Diseases ; microbiology ; Plants, Genetically Modified ; genetics ; microbiology ; Virulence ; Xanthomonas ; genetics ; pathogenicity ; physiology

OBJECTIVETo transform HPV E6/E7 immortalized human oral epithelial cell (HIOEC) line cells by benzo(a)pyrene [B(a)P] in vitro, and to establish a carcinogenesis model of oral squamous cell carcinoma.

METHODSHIOEC cells were treated with 0.1 mg/L-1.2 mg/L B(a)P for 6 months. The cells were cloned at 18th passage, and then the culture medium was changed into DMEM containing 10% FBS at 21th passage. Cells were cultured in vitro for half and one year and the cell line was named HIOEC-B(a)P. The morphological changes of the cells were observed with differential interference contrast microscope and HE staining. The soft agar colony forming ability and tumorigenicity of the cells in nude mice were identified to confirm the malignant characteristics of HIOEC-B(a)P cells.

RESULTS(1) After HIOEC cells were treated with B(a)P for 6 months, HIOEC-B(a)P cells could grow well in DMEM medium containing 10% FBS and physical concentration of calcium. (2) When HIOEC cells were treated with chemical carcinogens, the morphology of the cells was changed. Cells showed the character of polygon epithelial cells with much atypical mitosis. (3) The 93th passage of HIOEC-B(a)P cells had soft agar colony formation ability. (4) The 55th passage of HIOEC-B(a)P cells could develop parakeratosis mass. The 69th passage of HIOEC-B(a)P cells could develop typical well-differentiated squamous cell carcinoma. The 74th and the 96th HIOEC-B(a)P cells developed I-II grade squamous cell carcinoma-like clinical lesions in nude mice.

CONCLUSIONSB(a)P may induce HIOEC cells to be oral squamous cell carcinoma (OSCC) carcinogenetic cells. It will provide a multiple factors, multistage carcinogenesis model of OSCC for the further research.

Animals ; Benzo(a)pyrene ; toxicity ; Carcinoma, Squamous Cell ; chemically induced ; pathology ; virology ; Cell Line, Transformed ; Cell Line, Tumor ; Cell Transformation, Viral ; Epithelial Cells ; pathology ; Human papillomavirus 16 ; genetics ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; chemically induced ; pathology ; virology ; Neoplasms, Experimental

OBJECTIVETo investigate the expression of galectin-1 in oral squamous cell carcinoma(OSCC) and its clinical significance.

METHODSDetection of the mRNA and protein expression of galectin-1 in the in vitro cellular carcinogenesis model of OSCC, OSCC cell lines and tissue specimens from 30 primary OSCC patients were performed using real-time polymerase chain reaction (PCR), Western blotting and immunohistochemistry, respectively.

RESULTSThe value of galectin-1 mRNA and protein level in human immortalized oral epithelia cell (HIOEC) cell was 0.071 ± 0.023, 0.118 ± 0.046, Compared with the HIOEC, galectin-1 mRNA level and protein expression were increased significantly in all the cell lines (0.141 ± 0.049, 0.504 ± 0.33) (P < 0.01). The levels of mRNA and protein expression of galectin-1 were significantly higher in the cancerous tissue (0.059 ± 0.034, 1.5 ± 0.68) than in the normal adjacent tissues (0.029 ± 0.012, 0.4 ± 0.56) (P < 0.01).

CONCLUSIONSThe expression of galectin-1 gene up-regulated in carcinogenesis process of OSCC significantly may be related to the tumorigenesis and development of OSCC, which illustrates its potential clinical application as tumor marker for early diagnosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Transformed ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Epithelial Cells ; cytology ; metabolism ; Female ; Galectin 1 ; genetics ; metabolism ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; pathology ; Mouth Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Up-Regulation

BACKGROUNDThe present study was designed to examine and analyze the global gene expression changes during the tumorigenesis of a human immortalized oral epithelial cell line, and search for the possible genes that may play a role in the carcinogenesis of oral cancer associated with benzo (a) pyrene.

METHODSThe human immortalized oral epithelial cells, which have been established through transfection of E6/E7 genes of human papillomavirus type 16 and proved to be non-tumorigenic in nude mice, were treated with benzo (a) pyrene. Tumorigenicity of the treated cells were examined through nude mice subcutaneous injection. The global gene expression profiles of immortalized cells and the tumorigenic cells were acquired through hybridization of a microarray of Affymetrix U133 plus 2.0. The data were analyzed using Spring 7.0 software and treated statistically using one-way analysis of variance (ANOVA). The differentially expressed genes were classified using a Venn diagram and annotated with gene ontology. Several highlighted genes were validated in cells using a real-time polymerase chain reaction.

RESULTSThere were 883 differentially expressed genes during the tumorigenesis and most of them changed expression in the early stage of tumorigenesis. These genes mainly involved in macromolecule metabolism and signal transduction, possessed the molecular function of transition metal ion binding, nucleotide binding and kinase activity; their protein products were mainly integral to membranes or localized in the nucleus and cytoskeleton. The expression patterns of IGFBP3, S100A8, MAP2K, KRT6B, GDF15, MET were validated in cells using a real-time polymerase chain reaction; the expression of IGFBP3 was further validated in clinical oral cancer specimens.

CONCLUSIONSThis study provides the global transcription profiling associated with the tumorigenesis of oral epithelial cells exposed to benzo (a) pyrene; IGFBP3 may play a potential role in the initiation of oral cancer related to benzo (a) pyrene exposure.

Benzo(a)pyrene ; toxicity ; Cell Transformation, Neoplastic ; Cells, Cultured ; Connexin 43 ; genetics ; Gene Expression Profiling ; Growth Differentiation Factor 15 ; genetics ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor Binding Proteins ; genetics ; Mouth Neoplasms ; chemically induced ; metabolism ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo interfere in the Tca8113-CDDP cell line with siRNA of cyclin D1 and to investigate time and dose dependent gene silencing effect of siRNA of cyclin D1.

METHODSsiRNA of cyclin D1 was transfected into Tca8113-CDDP cells Fluorescent CY3 dye labeled siRNA GAPDH was used as the control. The transient transfecting efficiency was examined at 4, 24, 48 and 72 h. The relative quantity of the target RNA of cyclin D1 was analyzed with SYBR Green fluorescent dye kit by the Real-time PCR assay. The protein level of cyclin D1 was examined with Western blot. The changes of cisplatin sensitivity after treatment with siRNA cyclin D1 were examined with methyl thiazolyl tetrazolium (MTT) assay.

RESULTSThe optimized transfecting efficiency with CY3 labeled siRNA GAPDH in Tca8113-CDDP cells was over 90%. The silencing rate of cyclin D1 siRNA was 81.6% at 24 h, 80.7% at 48 h and 94.3% at 72 h. Dose-dependent manner of gene silencing effect was observed when the siRNA concentration was lower than 100 nmol/L, however, gene silencing effect reached its platform when the concentration was higher than 100 nmol/L. The protein levels of cyclin D1 at 24, 48 and 72 h after transfection decreased significantly, and so did the growth of cells. Inhibition rates of cell growth induced by cisplatin after administration with or without cyclin D1 siRNA were 58.4% and 34.8%, respectively.

CONCLUSIONSChemical synthesized cyclin D1 siRNA effectively silenced the expression of cyclin D1 gene in Tca8113-CDDP cells in vitro, with a time- and dose-dependent manner and target gene silence in cells increased its sensitivity to cisplatin.

Antineoplastic Agents ; pharmacology ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Cyclin D1 ; genetics ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Neoplastic ; drug effects ; Gene Silencing ; Humans ; RNA, Small Interfering ; pharmacology ; Tongue Neoplasms ; genetics ; metabolism ; Transfection

OBJECTIVETo clarify the relationship between cyclin D1 and cisplatin resistance of Tca8113/cis diamminedichloroplatinum (CDDP) in vitro and in vivo.

METHODSWe applied the transfection method with plasmids pcDNA3.1-antisense-cyclin D1 by Lipofectamine 2000. Tca8113/CDDP cells were used as control. MTT assay was used to identify the proliferation and sensibility of those cells to cisplatin. Subsequently, 18 nude mice were subcutaneously injected by those cells and divided into 3 groups with 6 mice in each group. Every mouse was treated by cisplatin with 5 mg . kg(-1) . d(-1) for 5 days. The sizes of tumor were measured every other day and were described with the growth curves. After 20 days, tumors were anatomized and weighed. The tumor inhibition ratios were calculated and the HE slides were observed to determine the cell sensibility to cisplatin.

RESULTSThe transfected cells with pcDNA3.1-antisense-cyclin D1grew more slowly than other cells and showed higher sensibility to cisplatin in vitro. The tumors developed by cells with pcDNA3.1-antisense-cyclin D1 were smaller than the The tumor inhibition ratio was 74% (P < 0.05). The necrosis area was larger in the tumors developed by the transfected cells with pcDNA3.1-antisense-cyclin D1 than other groups.

CONCLUSIONSAntisense oligonucleotides of cyclin D1 can improve the sensibility of Tca8113/CDDP cells to cisplatin and inhibit the growth of tumors.

Animals ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Cyclin D1 ; genetics ; Drug Resistance, Neoplasm ; drug effects ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mouth Neoplasms ; drug therapy ; pathology ; Neoplasms, Squamous Cell ; drug therapy ; pathology ; Oligonucleotides, Antisense ; genetics

Toxoplasma gondii is a serious foodborne zoonosis,which can not only affect the development of animal husbandry and the safety of meat products,but also cause great harm to public health.Therefore,the prevention of toxoplasmosis is crucial.Toxoplasma gondii vaccine which is regarded as an important measure to prevent toxoplasmosis has significant value to both public health and economics.This paper mainly summarizes advances in the study of Toxoplasma gondii vaccine in order to provide references for the further development of it,so as to control the toxoplasmosis more effectively.

Severe periodontitis is the main cause of tooth loss in adults, with varying degrees of horizontal and vertical alveolar bone loss. In view of the complex alveolar bone defect, a suitable surgery planning should be made on the basis of fully nuderstanding the characteristics of alveolar bone defect in severe periodontitis and the key points of bone augmentation technique, so as to choose an appropriate method for reconstruction of alveolar bone and complete the implantation and restoration to ensure the integrity of dentition, which are important for the long-term stability of periodontal health. Based on clinical experiences and literature review, we summarizes the characteristics of alveolar bone loss in patients with severe periodontitis and the timing of implant placement after bone augmentation surgery, in order to provide reference for implant treatment of severe periodontitis.