P63 and Ki-67 expression in radicular cyst.

Objectives: The aim of the current study was to identify the expression of P63 and its relation to odontogenic epithelial cell proliferation, severity of the inflammatory infiltrate and size of radicular cysts (RCs). Methods: In this retrospective cross-sectional study, 30 cases of paraffin-embedded RCs were randomly selected from the archive. P63 and Ki-67 expression was assessed by immunohistochemistry. Results: Epithelial P63 expression was absent in four (13.3%), weak in 10 (33.3%), and moderate in 16 (53.3%) cases. In the connective tissue wall of RC, P63 expression was absent in two (6.7%) cases, weak in 24 (80.0%) cases, and moderate in four (13.3%) cases. Ki-67 was found to be weakly expressed in 12 (40.0%) cases, moderately expressed in 13 (43.3%), and strongly expressed in five (16.7%) cases. No correlation was found between Ki-67 expression in odontogenic epithelium and P63 expression in the odontogenic epithelium (rho = 0.110, p = .563) or fibrous capsule (rho = 0.160, p = .399). Nevertheless, we found a positive correlation between Ki-67 expression in the odontogenic epithelium and the size of the RC (rho = 0.450, p = .013). The inflammatory infiltrate was negatively correlated with P63 expression in the odontogenic epithelium (rho = -0.428, p = .018), and with the size of cysts (rho = -0.728, p < .001). Conclusions: There is a high expression of P63 throughout the odontogenic epithelium and connective tissue capsule of the RC. P63 expression in the odontogenic epithelium is negatively correlated with the degree of the inflammatory infiltrate but not with epithelial cell proliferation or the size of the cyst.

The Prevalence of EBV and KSHV in Odontogenic Lesions.

OBJECTIVES: Odontogenic lesions evolve as a result of altered dental development. This study aimed to evaluate the prevalence and the coinfection of Epstein-Barr virus (EBV) and Kaposi sarcoma-associated herpesvirus (KSHV) in radicular cysts, dentigerous cysts, odontogenic keratocysts, and ameloblastomas. METHODS: Polymerase chain reaction (PCR) was used to analyse 66 cases of odontogenic lesions for the presence of EBV-DNA and KSHV-DNA. These lesions were 15 radicular cysts, 16 dentigerous cysts, 18 odontogenic keratocysts, and 17 ameloblastomas. RESULTS: EBV-DNA was detected in 24 (36.4%) of the studied samples as follows: 6 samples (40.0%) of radicular cysts, 4 (25.0%) of dentigerous cysts, 10 (55.6 %) of odontogenic keratocysts, and 4 (23.5%) of ameloblastomas (P = .168). KSHV-DNA was found in 16 (24.2%) of the studied samples as follows: 1 sample (6.7%) of radicular cysts, 6 (37.5%) of dentigerous cysts, 8 (44.4 %) of odontogenic keratocysts, and 1 (5.9%) of ameloblastomas (P = .001). Additionally, EBV and KSHV were positively correlated in all studied samples (P = .002). CONCLUSIONS: Both EBV and KSHV are found in odontogenic cysts and ameloblastomas. KSHV and EBV are more prevalent in odontogenic keratocysts than in other studied odontogenic lesions. Further, there is a high prevalence of EBV and KSHV coinfection in odontogenic cysts and ameloblastomas.

The correlated expression of COX-2 and keratin 15 in radicular cysts.

Background: The expression of cyclooxygenase-2 (COX-2) and Keratin-15 (K15) in radicular cysts (RCs) is poorly understood. Identifying the expression of these two markers may modify the current treatment of RC. The objective of this study was to evaluate the expression of COX-2 and its relationship to K15 expression in the odontogenic epithelial cells of the RC. Material and Methods: A total of 18 RCs were immunohistochemically analyzed for COX-2 and K15 expression. The cellular inflammatory reaction in the cyst wall was also assessed by measuring the percentage of inflammatory cells to the total number of cells. Results: COX-2 expression in the odontogenic epithelium of RC was absent in 11.1 % (n=2), mild in 27.8 % (n=5), moderate in 22.2% (n=4) and strong in 38.9% (n=7). Meanwhile, K15 expression was absent in 27.8% (n=5), mild in 16.7% (n=3), moderate in 44.4% (n=8), and strong in 11.1% (n=2) of the cases. The inflammatory infiltrate was mild in 2 cases (11.1%), moderate in 6 cases (33.3%), and high in 10 cases (55.6%). Spearman's correlation test revealed significant correlation (rho= .533; p= .023) between COX-2 and K15 expression in the odontogenic epithelium of RC. However, no correlation was noted between inflammation and expression of COX-2 (rho= 0.248, p=.321) or K15 (rho= -0.162, p= .520). Conclusions: There is high and correlated expression of COX-2 and K15 in the odontogenic epithelium of RC. COX-2 could therefore be involved in epithelial cell differentiation of the cyst. Additionally, the expression of K15 in RC may be an indicator of epithelial cell differentiation. Key words:Cyclooxygenase, COX-2, Keratin-15, K15, Radicular cyst.

[The expression and clinical significance of Bmi-1 gene in oral leukoplakia with different prognosis].

Objective: To detect the expression of Bmi-1 in oral leukoplakia (OL) cells and tissues, and analyze its role and clinical significance in the malignant transformation of OL. Methods: Immunohistochemistry was used to evaluate Bmi-1 expression in OL samples from 109 patients (51 males, 58 females, age range: 18-74 years) who were treated in the Department of Stomatology, the Affiliated Wuxi People's Hospital of Nanjing Medical University and the Center of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, between 1996 and 2018. The correlation between Bmi-1 expression level and clinicopathological parameters and prognosis in patients with OL was analyzed. The mRNA and protein expression levels of Bmi-1 gene in normal oral mucosal epithelial cells, OL cells, oral squamous cell carcinoma (OSCC) cells, OL tissues and paired adjacent normal tissues were detected by real-time PCR and Western blotting. The effects of Bmi-1 on the proliferation, colony formation and apoptosis were investigated by silencing expression of Bmi-1 in OL cell lines Leuk-1. Results: The protein level of Bmi-1 in OL tissue with severe and mild dysplasia was statistically different (6 819±994 vs 4 713±372, P=0.017). The OSCC-free survival rate of OL patients with high Bmi-1 expression was 65.5% (36/55), which was lower than that of OL patients with low Bmi-1 expression (88.9%, 48/54, P=0.003). Multivariate Cox proportional analysis indicated that Bmi-1 expression was the independent predictor for malignant transformation of OL (HR=2.522, 95%CI: 1.128-5.640, P=0.024). The mRNA level of Bmi-1 in OL specimens was 0.455±0.120, which was higher than that in paired adjacent normal tissues (0.063±0.009, P=0.014). The Bmi-1 mRNA level in malignant transformed OL specimens was (1.405±0.397), which was higher than that in untransformed OL specimens (0.145±0.017, P<0.001). After transfection of Bmi-1-shNC and Bmi-1-shRNA2 adenovirus into OL cell line Leuk-1, there were significant differences in the number of clone formation (824±40 vs 414±38, P=0.002) and apoptosis rate (17.7%±2.3% vs 36.0%±2.0%, P=0.004). Conclusions: The up-regulation of Bmi-1 expression promotes the malignant biological behavior of OL cells. Bmi-1 expression can be used as a predictor for malignant transformation of OL.

Human cytomegalovirus is present in the odontogenic epithelium of ameloblastoma.

Background: The factor behind the activation of the remnant odontogenic tissues and development of odontogenic cysts and tumors is poorly understood.This study aimed to investigate the presence of human cytomegalovirus (HCMV) in dentigerous cyst (DC), odontogenic keratocyst (OKC), and ameloblastoma (AB). Methods: The study included 41 samples, which distributed into DC (n=13), OKC (n=12), and AB (n=16). Conventional PCR assay and IHC analysis were used to detect the HCMV-DNA and HCMV glycoprotein B (HCMV-gB) respectively. Results: HCMV-DNA was detected in 10 samples (62.5%) of AB, four samples (30.8%) of DC, and three samples (25 %) of OKC respectively (χ2 test = 1.195, p= 0.247). Meanwhile, HCMV-gB was found in 12 (75%) of AB, in 2 (15.4%) of DC, and absent in OKC (0.0%) (χ2 test = 4.122, p= 0.042). Conclusions: The high prevalence of HCMV inside the odontogenic epithelium of AB could indicate a possible role of the virus in the oncogenesis and/or oncomodulation of the AB. Additionally, we recommend the IHC for the detection of HCMV in the odontogenic tumors like AB.

Accuracy and Effects of Clinical Decision Support Systems Integrated With BMJ Best Practice-Aided Diagnosis: Interrupted Time Series Study.

BACKGROUND: Clinical decision support systems (CDSS) are an integral component of health information technologies and can assist disease interpretation, diagnosis, treatment, and prognosis. However, the utility of CDSS in the clinic remains controversial. OBJECTIVE: The aim is to assess the effects of CDSS integrated with British Medical Journal (BMJ) Best Practice-aided diagnosis in real-world research. METHODS: This was a retrospective, longitudinal observational study using routinely collected clinical diagnosis data from electronic medical records. A total of 34,113 hospitalized patient records were successively selected from December 2016 to February 2019 in six clinical departments. The diagnostic accuracy of the CDSS was verified before its implementation. A self-controlled comparison was then applied to detect the effects of CDSS implementation. Multivariable logistic regression and single-group interrupted time series analysis were used to explore the effects of CDSS. The sensitivity analysis was conducted using the subgroup data from January 2018 to February 2019. RESULTS: The total accuracy rates of the recommended diagnosis from CDSS were 75.46% in the first-rank diagnosis, 83.94% in the top-2 diagnosis, and 87.53% in the top-3 diagnosis in the data before CDSS implementation. Higher consistency was observed between admission and discharge diagnoses, shorter confirmed diagnosis times, and shorter hospitalization days after the CDSS implementation (all P<.001). Multivariable logistic regression analysis showed that the consistency rates after CDSS implementation (OR 1.078, 95% CI 1.015-1.144) and the proportion of hospitalization time 7 days or less (OR 1.688, 95% CI 1.592-1.789) both increased. The interrupted time series analysis showed that the consistency rates significantly increased by 6.722% (95% CI 2.433%-11.012%, P=.002) after CDSS implementation. The proportion of hospitalization time 7 days or less significantly increased by 7.837% (95% CI 1.798%-13.876%, P=.01). Similar results were obtained in the subgroup analysis. CONCLUSIONS: The CDSS integrated with BMJ Best Practice improved the accuracy of clinicians' diagnoses. Shorter confirmed diagnosis times and hospitalization days were also found to be associated with CDSS implementation in retrospective real-world studies. These findings highlight the utility of artificial intelligence-based CDSS to improve diagnosis efficiency, but these results require confirmation in future randomized controlled trials.

p63 Expression and its Relation to Epithelial Cells Proliferation in Dentigerous Cyst, Odontogenic Keratocyst, and Ameloblastoma.

The current controversy about the classification of odontogenic keratocyst reflects the shortage in the understanding of the odontogenic cysts and tumors. The aim of the present study was to investigate p63 immunoexpression and its relation to the proliferation of the epithelial lining in dentigerous cyst (DC), odontogenic keratocyst (OKC), and follicular type of ameloblastoma (AB). The study involved 36 samples, which are DC (n = 12), OKC (n = 9), and AB (n = 15). p63 protein expression was evaluated by immunohistochemistry. The results on Ki-67 expression were obtained from our previous studies and correlated with p63 expressions. p63 was expressed differently in the studied lesions with various distribution in different study samples. Statistical analysis using Kruskal-Wallis test showed a significant difference in the expression of p63 protein among DC, OKC, and AB (p = 0.048). Subsequently, Mann-Whitney U test revealed the expression of p63 protein was significantly higher in OKC than DC (p = 0.018). Interestingly, Spearman's correlation analysis showed a positive correlation between the expression of p63 and Ki-67 in the odontogenic epithelium of DC (σ = 0.757, P = 0.004) and OKC (σ = 0.741, P = 0.022). While no such a positive correlation was found between the two studied markers in AB group (σ = 0.006, P = 0.983). In conclusion, the present results indicated various expression and correlation of p63 with the proliferation of odontogenic epithelial cells in DC, OKC, and AB. This diversity could reflect a different role and pathways of ΔNp63 in odontogenic tumor than that in odontogenic cyst. These together will help in better understanding the pathogenesis and biological behavior of odontogenic cysts and tumors.

Expression of keratin 15 in dentigerous cyst, odontogenic keratocyst and ameloblastoma.

The etiology and pathogenesis of odontogenic lesions are poorly understood. Keratin 15 (K15) is a type I cytoskeletal protein that provides structural support to the cells and has been considered to be a stem cell marker. The aim of the present study was to evaluate the expression of K15 in the epithelial lining of dentigerous cysts (DCs), odontogenic keratocysts (OKCs) and ameloblastomas (ABs). The study included 41 samples of DCs (n=13), OKCs (n=12), and AB tissues (n=16). K15 protein expression was evaluated via immunohistochemistry and data were statistically analyzed using a Kruskal-Wallis test. K15 was expressed in the majority of the studied lesions with various distributions in the different study samples. The Kruskal-Wallis test revealed non-significant differences in the expression of K15 among the three odontogenic lesions (P=0.380). The present study confirmed the high expression of K15 in the different epithelial layers of DC, OKC and AB. This type of expression excludes the reliability of regarding K15 as a stem cell marker in DC, OKC and AB. However, K15 may reflect the abnormal differentiation of pathological epithelial cells in these lesions.

The Efficacy and Safety of Miconazole Nitrate Mucoadhesive Tablets versus Itraconazole Capsules in the Treatment of Oral Candidiasis: An Open-Label, Randomized, Multicenter Trial.

BACKGROUND: Oral candidiasis (OC) is a common oral fungal infection. Recently, miconazole mucoadhesive tablets have been gaining attention for OC treatment. Despite trials in patients with human immunodeficiency virus and cancer, evidence of its application in the large-scale, general population with OC is lacking. This study aimed to evaluate the efficacy and safety of miconazole nitrate mucoadhesive tablets in comparison with itraconazole capsules for OC treatment. METHODS: The study was a randomized, open-label, parallel-armed, multicenter clinical trial. Totally, 343 patients diagnosed with OC, who met the inclusion criteria, were randomly assigned to either a treatment group that received miconazole nitrate mucoadhesive tablets (10 mg) once daily or a control group that received itraconazole capsules (100 mg QD) for 2 weeks, and were followed up for 2 weeks. The clinical cure, improvement of clinical symptoms/signs, mycologic cure, and safety were evaluated. RESULTS: The mucoadhesive tablets (n = 171) did not show inferiority to itraconazole (n = 172) in the treatment of OC. At the end of the 14-day treatment, the clinical cure rates were 45.29% and 41.76% in the miconazole and itraconazole groups, respectively (P = 0.3472). At the end of the 14-day follow-up, the clinical cure rates were 51.18% and 41.76% in the miconazole and itraconazole groups, respectively (P = 0.0329). Adverse events occurred in 53 subjects (33 in the miconazole group and 20 in the itraconazole group). There was no statistical difference in the safety profile between miconazole and itraconazole (P = 0.0533). Thrombocytopenic purpura, although rare, occurred in one patient in the miconazole group and was considered a drug-related, severe adverse event. CONCLUSION: Miconazole nitrate mucoadhesive tablets may be as effective as systemic itraconazole capsule for OC treatment. Physicians should be cautious about thrombocytopenic purpura occurring as a rare and serious adverse event of miconazole nitrate. TRIAL REGISTRATION: Chinese Clinical Trial Register ChiCTR-TRC-13003935.

Salivary Heparanase Level Is a Potential Biomarker to Diagnose and Prognose the Malignant Salivary Gland Tumor.

BACKGROUND: Upregulation of heparanase has been reported in an increasing number of human cancer tissues. However, the level of salivary heparanase and its clinical significance in patients with salivary gland tumors remain unclear. METHODS: Salivary heparanase levels in patients with salivary gland tumors were detected using enzyme-linked immunosorbent assays (ELISAs) and the clinical significance was evaluated by analyzing the correlations among salivary heparanase levels, clinicopathological parameters, and clinical outcomes. RESULTS: The levels of salivary heparanase were significantly higher in patients with malignant salivary gland tumors than in benign tumors and normal controls (P<0.0001). High salivary heparanase levels were positively correlated with increased lymph node metastasis (P = 0.0235) and poorer tumor node metastasis stage (TNM) (P = 0.0183). Survival analyses revealed that high salivary heparanase levels were associated with worse overall survival (P = 0.0023) and disease-free survival (DFS) (P = 0.0025). CONCLUSIONS: The study shows that salivary heparanase levels, as detected by the ELISAs, can be used to diagnose and provide an accurate prognosis for malignant salivary gland tumors. Salivary heparanase level was an independent predictor in patients with malignant salivary gland tumors.

Estimation of the binding modes with important human cytochrome P450 enzymes, drug interaction potential, pharmacokinetics, and hepatotoxicity of ginger components using molecular docking, computational, and pharmacokinetic modeling studies.

Ginger is one of the most commonly used herbal medicines for the treatment of numerous ailments and improvement of body functions. It may be used in combination with prescribed drugs. The coadministration of ginger with therapeutic drugs raises a concern of potential deleterious drug interactions via the modulation of the expression and/or activity of drug-metabolizing enzymes and drug transporters, resulting in unfavorable therapeutic outcomes. This study aimed to determine the molecular interactions between 12 main active ginger components (6-gingerol, 8-gingerol, 10-gingerol, 6-shogaol, 8-shogaol, 10-shogaol, ar-curcumene, ß-bisabolene, ß-sesquiphelandrene, 6-gingerdione, (-)-zingiberene, and methyl-6-isogingerol) and human cytochrome P450 (CYP) 1A2, 2C9, 2C19, 2D6, and 3A4 and to predict the absorption, distribution, metabolism, excretion, and toxicity (ADMET) of the 12 ginger components using computational approaches and comprehensive literature search. Docking studies showed that ginger components interacted with a panel of amino acids in the active sites of CYP1A2, 2C9, 2C19, 2D6, and 3A4 mainly through hydrogen bond formation, to a lesser extent, via π-π stacking. The pharmacokinetic simulation studies showed that the [I]/[Ki ] value for CYP2C9, 2C19, and 3A4 ranged from 0.0002 to 19.6 and the R value ranged from 1.0002 to 20.6 and that ginger might exhibit a high risk of drug interaction via inhibition of the activity of human CYP2C9 and CYP3A4, but a low risk of drug interaction toward CYP2C19-mediated drug metabolism. Furthermore, it has been evaluated that the 12 ginger components possessed a favorable ADMET profiles with regard to the solubility, absorption, permeability across the blood-brain barrier, interactions with CYP2D6, hepatotoxicity, and plasma protein binding. The validation results showed that there was no remarkable effect of ginger on the metabolism of warfarin in humans, whereas concurrent use of ginger and nifedipine exhibited a synergistic effect on platelet aggregation in humans. Moreover, ginger components showed a rapid half-life and no to low toxicity in humans. Taken together, this study shows that ginger components may regulate the activity and expression of various human CYPs, probably resulting in alterations in drug clearance and response. More studies are warranted to identify and confirm potential ginger-drug interactions and explore possible interactions of ginger with human CYPs and other functionally important proteins, to reduce and avoid side effects induced by unfavorable ginger-drug interactions.

[Expressions of RORγT; and FOXP3 and clinical significance in patients with oral lichen planus].

PURPOSE: Oral lichen planus was considered as T cell mediated autoimmune disease affecting oral mucosa with unknown etiopathogenesis. Helper T lymphocytes played an important role in the pathogenesis of OLP. The purpose of this study was to investigate the possible role of Th17 and Treg cells in OLP lesions. METHODS: Forty three patients with OLP (24 patients with reticular OLP and 19 patients with atrophic-erosive OLP) and 13 healthy volunteers were recruited in this study. Real-time quantitative PCR was performed to detect the expressions of RORRORγT and FOXP3, which were the lineage-specific transcription factors for Th17 and Treg, respectively. Statistical difference was evaluated by GraphPad Prism 5 software. RESULTS: The results showed that the expressions of RORRORγT and FOXP3 in OLP lesions were significantly higher than that in normal oral mucosa, and correlated with the clinical classification of OLP. Additionally, it was found that RORRORγT/FOXP3 ratio in atrophic-erosive OLP was significantly higher than that in reticular OLP and control group; Moreover, increased RORRORγT/FOXP3 ratio in reticular OLP was found compared with control group, but the difference was not statistically significant. CONCLUSIONS: The results demonstrate that Th17 and Treg cells participate in the immune response in OLP lesions. Th17 predominance of Th17/Treg imbalance may implicate the immune response in atrophic-erosive OLP. These findings help to broaden our view on the pathogenesis of OLP.

Implications of Th1 and Th17 cells in pathogenesis of oral lichen planus.

Oral lichen planus (OLP) is considered a T cell-mediated autoimmune disease with unknown aetiology. T helper cells appear to play an important role in the pathogenesis of OLP. We investigated the possible role of T helper cells, Th1 and Th17, in the lesions and circulation of patients with OLP. Forty patients with OLP and 15 healthy volunteers were recruited. Double immunofluorescence staining was used to detect Th1 and Th17 cells in the OLP lesions, and intracellular cytokine staining and flow cytometry to evaluate the proportion of Th1 and Th17 cells in peripheral blood. The levels of serum interferon (IFN)-γ and interleukin (IL)-17 were assessed by using an enzyme-linked immunosorbent assay. It was found that Th17 cells, as well as Th1 cells, were present in OLP lesions. The proportion of peripheral Th1 and Th17 cells was significantly increased in patients with OLP. The proportion of Th17 cells in atrophic-erosive OLP was elevated as compared with that in reticular OLP. Serum IL-17 levels in OLP patients were significantly higher than in controls, and those in the atrophic-erosive OLP group were increased as compared with the reticular OLP group. However, the levels of serum IFN-γ were slightly decreased in OLP patients. Our data suggested that Th1 and Th17 cells in the local lesions and peripheral blood may be associated with the pathogenesis of OLP, and that IL-17 may be an important proinflammatory cytokine in OLP. These findings enhance our understanding of OLP pathogenesis.

Correlation between distribution of Helicobacter pylori in oral cavity and chronic stomach conditions.

In this study, the colonization and distribution of Helicobacter pylori (Hp) in patients with chronic gastric diseases were investigated and the relationship between the periodontal initial treatment and presence of Hp in oral cavity was examined to better understand the connection between Hp infection and chronic diseases. Primers for PCR amplification were designed according to ureC gene and cagA genes of Hp. Specimens were harvested from different sites of 96 patients with chronic gastric diseases and the specimens of dental plaques, gargles and dorsal mucosa were tested for Hp. The 96 patients were treated by bismuth triple therapy and among them, 52 subjects were additionally given periodontal initial therapy. The eradication rate of gastric Hp and oral Hp detection rate were determined 4 weeks and 1 year after the treatment. The results showed that the detection rates of oral specimens were in the order of dental plaques (82.3%), gargles (51.1%) and scrapings of dorsal mucosa of tongue (37.5%). One year after bismuth triple therapy or the triple therapy in combination with periodontal initial treatment, the eradication rate of gastric Hp was significantly higher in the combination treatment group than in group treated by the triple therapy alone (62.8% vs. 32.4%, P<0. 05). Moreover, the Hp detection rate was significantly lower in the combination group than in the group treated only with the triple therapy. We are led to conclude that Hp is present at various parts of oral cavity, oral Hp might be an important source of gastric Hp and the triple therapy plus periodontal initial treatment can enhance the long-term eradication rate of gastric Hp in patient with both chronic gastric diseases and chronic periodontitis.

Correlation between Distribution of Helicobacter Pylori in Oral Cavity and Chronic Stomach Conditions / 华中科技大学学报（医学）（英德文版）

In this study,the colonization and distribution of Helicobacter pylori (Hp) in patients with chronic gastric diseases were investigated and the relationship between the periodontal initial treatment and presence of Hp in oral cavity was examined to better understand the connection between Hp infection and chronic diseases.Primers for PCR amplification were designed according to ureC gene and cagA genes of Hp.Specimens were harvested from different sites of 96 patients with chronic gastric diseases and the specimens of dental plaques,gargles and dorsal mucosa were tested for Hp.The 96 patients were treated by bismuth triple therapy and among them,52 subjects were additionally given periodontal initial therapy.The eradication rate of gastric Hp and oral Hp detection rate were determined 4 weeks and 1 year after the treatment.The results showed that the detection rates of oral specimens were in the order of dental plaques (82.3％),gargles (51.1％) and scrapings of dorsal mucosa of tongue (37.5％).One year after bismuth triple therapy or the triple therapy in com bination with periodontal initial treatment,the eradication rate of gastric Hp was significantly higher in the combination treatment group than in group treated by the triple therapy alone (62.8％ vs.32.4％,P＜0.05).Moreover,the Hp detection rate was significantly lower in the combination group than in the group treated only with the triple therapy.We are led to conclude that Hp is present at various parts of oral cavity,oral Hp might be an important source of gastric Hp and the triple therapy plus periodontal initial treatment can enhance the long-term eradication rate of gastric Hp in patient with both chronic gastric diseases and chronic periodontitis.

[The level of PPTA mRNA and its proteins expression in trigeminal ganglia in rats with occlusal recovery].

PURPOSE: To study the level of substance P(SP) and preprotachykinin A(PPTA) mRNA in trigeminal ganglia(TG)in rats with occlusal recoveryìand to discuss the mechanism of temporomandibular disorders. METHODS: 48 Wistar male rats were randomly divided into 3 experimental groups and 3 control groupsì8 rats in each group. The right maxillary and mandibular molars in the experimental groups were ground to the gingival level without occlusal contact. The occlusal contact was recovered by stoping grinding the molars gradually. The section of trigeminal ganglia were used for immunohistological and in situ hybridization study. Light microscope and microscoic photo analytic software were employed to detect the percentage of SP and PPTA mRNA positive neurons in the frozen section of TGs in 48 rats. SPSS10.0 software package were used for statistical analysis. RESULTS: The percentage of SP positive neurons in TG with unilateral chew experimental group decreased significantly compared to the control group (P<0.01,P<0.05).The decreased extent in the non-chewing side was much higher than that in the chewing side(P<0.05).The percentage of PPTA mRNA positive neurons was significantly higher in both of the chewing and non-chewing sides of the unilateral chew experimental group than that in the control group (P<0.05,P<0.01)ì and that non-chewing side was significantly higher than that in the chewing side(P<0.01).There was no significant difference in the percentage of SP and PPTA mRNA positive neurons between the early occlusal reconstruction the experimental group and the control group(P>0.05), and there was no significant difference of those between the non-chewing side and the chewing side(P>0.05). The results of later occlusal recovery in the experimental group was same to that of the unilateral chew experimental group. CONCLUSIONS: The expressions of SP and PPTA mRNA in TG can recover normal in the early occlusal recovery but can not in the later occlusal recovery. SP and PPTA mRNA might participate in the histopathologic mechanism of temporomandibular disorders.

Third-molar development in relation to chronologic age in young adults of central China.

The estimation of chronologic age based on the stages of third-molar development was evaluated by using the eight stages (A-H) method of Demirjian and the third-molar development was compared, in terms of sex and age, with results of previous studies. The samples consisted of 291 orthopantomograms from young Chinese subjects of known chronologic age and sex (including 139 males with a mean age of 14.67+/-3.62 y and 152 females with a mean age of 14.85+/-3.70 y). Statistical analysis was performed by employing the Mann-Whitney U-test and the t-test. Regression analysis was conducted to obtain regression formulas for calculating dental age from the chronologic age. Our results showed statistically significant differences (P<0.05) in third-molar development between males and females, at the calcification stages D, E and H. And a strong correlation was found between age and third-molar development in both males (r (2)=0.65) and females (r (2)=0.61). New equations (Age=8.76+1.32 Development stage) for estimating chronologic age were derived. It is concluded that third-molar genesis took place earlier in males than in females. The use of third molars as a developmental marker is appropriate in young adults of Central China. The formula obtained in the present study can be used as a guide for estimation of dental maturity and a standard for age estimation for young adults of Central China.

[Effects of basic fibroblast growth factor on the proliferation of human salivary adenoid cystic carcinoma cell line ACC-2 and extracellular signal-regulated kinase, Cyclin D1, p2waf/cip1 signaling pathway].

OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) on the proliferation of human salivary adenoid cystic carcinoma (ACC) cell line ACC-2 in vitro. RESULTS: The effect of ectogenic bFGF on proliferation of ACC-2 was observed by MTT assay. Extracellular signal-regulated kinase (ERK) activity was measured by immuno-precipitation. p-ERK1/2, Cyclin D1 and p21waf/cip1 expression were assessed by Western blot. RESULTS: bFGF could enhance the proliferation of ACC-2. Stimulated by bFGF, the proliferation ratio increased significantly. The intracellular ERK activity, p-ERK1/2 and Cyclin D1 expression were increased, while p21waf/cip1 expression was inhibited by different concentrations of bFGF. The above effects of bFGF could be attenuated by MEK inhibitor U0126. CONCLUSION: bFGF stimulates the proliferation of ACC-2 in a dose dependent manner. The proliferation effect of bFGF may be due to up-regulating ERK, Cyclin D1 and p21waf/cip1 signaling pathway. This research can help us to explore a new pathogenesis and therapy of the ACC.

Effect of exogenous bFGF on the proliferation of human adenoid cystic carcinoma ACC-2 cells.

To observe the effects of basic fibroblast growth factor (bFGF) on human adenoid cystic carcinoma ACC-2 cell line proliferation and ERK, cyclin D1/p21(waf/cip1) signaling pathways, human adenoid cystic carcinoma cells (ACC-2) were cultured and the influence of bFGF of different concentrations on cell proliferation was determined by MTT. Protein was detected by immuno-precipitation and ERK activity by using ERK agent kit. p-ERK(1/2) and down-stream cyclin D1, p21(waf/cip1) expression were detected by Western blotting and the interfering role of mitogen protein-activated kinase (MEK) suppressor U0126 in the afore-mentioned indicators was examined. MTT demonstrated ACC-2 cell proliferation was substantially enhanced by bFGF, immuno-precipitation displayed ERK activity was up-regulated by bFGF, and immuno-imprinting also showed p-ERK(1/2), cyclin D1 expression was greatly enhanced and p21(waf/cip1) expression was inhibited by bFGF. U0126 suppressed the effect of bFGF. It is concluded that bFGF can promote the proliferation of human adenoid cystic carcinoma ACC-2 cells, and its pathways are associated with the up-regulated activity and expression of p-ERK(1/2), inhibited p21waf/cip1 expression and enhanced cyclin D1 expression.

Third-Molar Development in Relation to Chronologic Age in Young Adults of Central China / 华中科技大学学报（医学）（英德文版）

The estimation of chronologic age based on the stages of third-molar development was evaluated by using the eight stages (A-H) method of Demirjian and the third-molar development was compared, in terms of sex and age, with results of previous studies. The samples consisted of 291orthopantomograms from young Chinese subjects of known chronologic age and sex (including 139males with a mean age of 14.67±3.62 y and 152 females with a mean age of 14.85±3.70 y). Statistical analysis was performed by employing the Mann-Whitney U-test and the t-test. Regression analysis was conducted to obtain regression formulas for calculating dental age from the chronologic age. Our results showed statistically significant differences (P＜0.05) in third-molar development between males and females, at the calcification stages D, E and H. And a strong correlation was found between age and third-molar development in both males (r2=0.65) and females (r2=0.61). New equations (Age=8.76+1.32 Development stage) for estimating chronologic age were derived. It is concluded that third-molar genesis took place earlier in males than in females. The use of third molars as a developmental marker is appropriate in young adults of Central China. The formula obtained in the present study can be used as a guide for estimation of dental maturity and a standard for age estimation for young adults of Central China.