The expression of Runx2 in the pathogenesis of periodontitis.

OBJECTIVE: Runt-related transcription factor 2 (Runx2) plays an important role in bone metabolism; however, the relationship between Runx2 and periodontitis remains unclear. We investigated Runx2 expression in the gingiva of patients to explore its role in periodontitis. METHODS: Gingival samples of patients were collected, including healthy samples (control group) and periodontitis samples (P group). Periodontitis samples were divided into three groups based on the periodontitis stage. Samples with stage I and grade B periodontitis were in the P1 group, stage II and grade B in the P2 group, and stage III or IV and grade B in the P3 group. Immunohistochemistry and western blotting were performed to detect Runx2 levels. The probing (PD) and clinical attachment loss (CAL) were recorded. RESULTS: Runx2 expression levels in the P and P3 groups were higher than those in the control group. In addition, Runx2 expression was positively correlated with CAL and PD (r1 = 0.435, r2 = 0.396). CONCLUSION: The high expression level of Runx2 in the gingiva of patients with periodontitis may be correlated with the pathogenesis of periodontitis.

Leptin regulates OPG and RANKL expression in Gingival Fibroblasts and Tissues of Chronic Periodontitis Patients.

Objective: Chronic periodontitis is a bone-destructive disease affecting periodontal support structures. Although leptin has a protective effect against periodontitis, the underlying mechanism remains unclear. Therefore, this study aimed to investigate the possible role of leptin by examining its relationship with OPG and RANKL in human gingival tissues obtained from patients with chronic periodontitis. Method: Twenty-two patients with chronic periodontitis were enrolled (10 with moderate periodontitis and 12 with severe periodontitis) in the experimental group, and 12 healthy individuals were enrolled in the control group. Gingival tissue samples were collected, and the protein levels and localization of leptin, OPG, and RANKL were studied using immunohistochemistry (IHC). The staining intensities of leptin, OPG, and RANKL were correlated with the periodontal clinical index. Moreover, real-time quantitative PCR (RT-qPCR) was used to determine OPG and RANKL mRNA levels in gingival fibroblasts stimulated with gradient concentrations of leptin protein in vitro. Result: Leptin, OPG, and RANKL were located in the cytoplasm of gingival epithelial cells and the connective tissue. Leptin was widely and significantly expressed in the control group, whereas it was lightly stained in the severe group. RANKL was lightly stained in the control group, whereas it was widely and significantly expressed in the severe group. The control and the moderate groups had similar OPG levels, which were significantly higher than that in the severe group. Leptin was positively correlated with OPG(r = 0.905, p < 0.01) and negatively correlated with RANKL (r = -0.635, p < 0.01). In vitro low concentrations of leptin led to an increased OPG/RANKL mRNA ratio, whereas the opposite effect was observed at high concentrations. Conclusion: Leptin can regulate OPG and RANKL expression in gingival fibroblasts and may thus play a role in the development of chronic periodontitis by modulating the OPG/RANKL ratio.

Expression and methylation of Dickkopf-1 in the pathogenesis and malignant transformation of oral submucous fibrosis.

BACKGROUND: Dickkopf-1 is an inhibitor of the Wnt/ß-catenin pathway, but the role of Dickkopf-1 in oral submucous fibrosis remains unclear. We evaluated the protein expression and gene methylation levels of dickkopf-1 to determine the mechanism underlying abnormal Wnt/ß-catenin pathway activation. METHODS: Healthy mucosa, oral submucous fibrosis, oral squamous cell carcinoma, and cancer-adjacent tissues were collected. The expression and promoter methylation levels of dickkopf-1 were analyzed. RESULTS: The expression levels of dickkopf-1 in oral submucous fibrosis and oral squamous cell carcinoma tissues were lower than those in healthy and cancer-adjacent tissues. The methylation levels of the dickkopf-1 gene in oral submucous fibrosis and oral squamous cell carcinoma tissues were higher than those in healthy and cancer-adjacent tissues. Dickkopf-1 expression was negatively correlated with dickkopf-1 gene methylation. CONCLUSIONS: High dickkopf-1 methylation levels in oral submucous fibrosis and oral squamous cell carcinoma tissues may decrease dickkopf-1 expression, which may induce an abnormal activation of the Wnt/ß-catenin pathway and oral submucous fibrosis pathogenesis.

[Hereditary gingival fibromatosis: a three-generation case report].

Hereditary gingival fibromatosis (HGF) is a familial hereditary disease; while it is rare and usually benign, it is also characterized by the slow and progressive development of gingival tissue. This paper reports on the clinical examina-tion and history of HGF in a family of patients.

[Clinical analysis for oral mucosal disease in 21 972 cases].

OBJECTIVE: To analyze the incidence and distribution of oral mucosal diseases in Hunan Province and provide reference for prevention and treatment.
 Methods: The clinical data for all patients, who were treated in Xiangya Hospital of Central South University from April 2013 to March 2017, were collected. After screening, weighing and classifying, sex and age distribution for the disease was analyzed.
 Results: The female with the age between 40 to 49 were in the majority among 21 972 patients. The ratio between men to women was 1:1.05. According to the classification of diseases, the most common diseases were as follows: recurrent aphthous ulcer (27.17%), burning mouth syndrome (15.72%), oral submucous fibrosis (14.75%), oral lichen planus (10.38%), oral leukoplakia (4.21%), traumatic ulceration (4.14%), chronic cheilitis (3.47%), oral fungal infection (3.26%), and atrophic glossitis (2.74%). Recurrent oral ulcer (28.65%), burning mouth syndrome (23.70%) and oral lichen planus (13.31%) were the most common 3 kinds of oral mucosal diseases during females in Hunan. Oral submucous fibrosis was the most common oral mucosal disease among males in Hunan (28.56%).
 Conclusion: Recurrent oral ulcer, burning mouth syndrome and oral lichen planus are very popular in women in Hunan Province, and oral submucous fibrosis is the most common disease in male in this region. It shows a high trend of incidence in the surrounding provinces.

[Expression and significance of secreted frizzled-related protein 1 and ß-catenin in gingival tissue of patients with chronic periodontitis].

OBJECTIVE: This study aimed to investigate the expression and correlation of secreted frizzled-related protein 1 (SFRP1) and ß-catenin in gingival tissues of patients with chronic periodontitis (CP). The role of the classical Wnt/ß-catenin signaling pathway in the development of periodontitis was also explored. METHODS: Twenty-eight patients with CP (CP group) were enrolled in this study. Among them, 16 cases were moderate CP, and 12 demonstrated severe CP. Twelve healthy cases comprised the controls (normal group). Gingival tissue was collected, and the probing depth, bleeding index, and clinical attachment loss were recorded. The expression levels of SFRP1 and ß-catenin were detected by immunohistochemistry, and staining intensity was evaluated by double scoring method. SPSS 19.0 was used for statistical analysis. RESULTS: The staining strength scores of SFRP1 and ß-catenin were 2.16±0.65 and 1.12±0.51 in the normal group, 3.57±0.45 and 2.36±0.49 in the CP group, 3.61±0.40 and 2.30±0.44 in the moderate CP group, and 3.52±0.52 and 2.45±0.55 in the severe CP group, respectively. The expression of SFRP1 and ß-catenin in the CP group was higher than that in the normal group (P<0.01). A significant difference was noted between the normal group and the moderate and severe CP groups (P<0.01) but none between the moderate and severe CP groups (P>0.05). A positive correlation was found between the expression of SFRP1 and ß-catenin (r=0.657, P<0.01). The expression levels of ß-catenin and SFRP1 were related to periodontal indexes. The correlation between the expression of SFRP1 and probing depth was most significant (r=0.723, P<0.01), as well as that between ß-catenin and bleeding index (r=0.697, P<0.01). CONCLUSIONS: Patients with CP exhibit elevated expression of SFRP1 and ß-catenin in gingival tissues, and this event is related to the degree of periodontal destruction. Abnormal expression of SFRP1 and ß-catenin may promote the development of periodontitis.

[Detection of Dickkopf-1 and alkaline phosphatase activity in gingival crevicular fluid from chronic periodontitis with Er:YAG laser as an adjunctive treatment].

PURPOSE: The aim of this study was to evaluate the clinical efficacy and the level of DKK1 and alkaline phosphatase (ALP) activity in gingival crevicular fluid (GCF) while taking Er:YAG laser as an adjunctive to scaling and root planning in the treatment of chronic periodontitis (CP). METHODS: Eleven patients with CP were included and there were nineteen pairs of homonym teeth(thirty-eight teeth) in this split-mouth design, and they were randomly assigned to experimental group or control group. In the experimental group, a combination of ultrasonic subgingigval scaling and root planning with hand instrument (SRP) were performed with Er: YAG laser as an adjunctive; in the control group, only SRP was performed. The main variables were bleeding index (BI), probing depth (PD), clinical attachment loss (CAL) which were assessed at baseline (1 week after ultrasonic subgingival scaling), l month and 3 months after treatment. GCF was collected at baseline, l week, l month and 3 months, and the levels of DKK1 and ALP activity were detected at the same time point. The data were analyzed with SPSS 19.0 software package. RESULTS: Both groups showed significant reduction of PD, CAL, BI values 1 month and 3months after treatment, but no significant difference in clinical parameters were found between the two groups. In the experimental group, the activity of ALP reduced to (386.69±146.42), (341.221±171.62), (249.27±98.72) from (396.191±150.55) U/L and the level of DKK1 dropped to (310.34±184.68), (270.04±55.14), (247.31±56.99) from (307.12±45.63) µg/L at the end of 1 week, 1 month, 3 months, respectively. Meanwhile, in the control group, the activity of ALP reduced to (374.72±131.27), (344.42±127.80), (252.36±90.4 ) from (394.09±120.25) U/L and the level of DKK1 dropped to (310.34±84.68), (270.04±55.14), (247.31±56.99) from (305.33±147.40) µg/L at the end of l week, l month, 3months, respectively. There is no significant difference between the two groups at any period for ALP or DKK1. CONCLUSIONS: Er:YAG laser was a safe no-surgical adjunctive therapy in treating chronic periodontitis, further observation is needed to determine its long-term effectiveness.

Quantitative proteomic analysis for novel biomarkers of buccal squamous cell carcinoma arising in background of oral submucous fibrosis.

BACKGROUND: In South and Southeast Asian, the majority of buccal squamous cell carcinoma (BSCC) can arise from oral submucous fibrosis (OSF). BSCCs develop in OSF that are often not completely resected, causing local relapse. The aim of our study was to find candidate protein biomarkers to detect OSF and predict prognosis in BSCCs by quantitative proteomics approaches. METHODS: We compared normal oral mucosa (NBM) and paired biopsies of BSCC and OSF by quantitative proteomics using isobaric tags for relative and absolute quantification (iTRAQ) to discover proteins with differential expression. Gene Ontology and KEGG networks were analyzed. The prognostic value of biomarkers was evaluated in 94 BSCCs accompanied with OSF. Significant associations were assessed by Kaplan-Meier survival and Cox-proportional hazards analysis. RESULTS: In total 30 proteins were identified with significantly different expression (false discovery rate < 0.05) among three tissues. Two consistently upregulated proteins, ANXA4 and FLNA, were validated. The disease-free survival was negatively associated with the expression of ANXA4 (hazard ratio, 3.4; P = 0.000), FLNA (hazard ratio, 2.1; P = 0.000) and their combination (hazard ratio, 8.8; P = 0.002) in BSCCs. CONCLUSION: The present study indicates that iTRAQ quantitative proteomics analysis for tissues of BSCC and OSF is a reliable strategy. A significantly up-regulated ANXA4 and FLNA could be not only candidate biomarkers for BSCC prognosis but also potential targets for its therapy.

[Effect of periodontal initial therapy on the amount of SFRP1 in gingival crevicular fluid in patients with periodontitis].

PURPOSE: To detect the change of secreted frizzled-related protein-1（SFRP1） in gingival crevicular fluid during periodontal initial treatment and explore the relationship between SFRP1 and the activity of chronic periodontitis. METHODS: Twenty-two patients with moderate to severe periodontitis were selected, and 5 healthy volunteers were enrolled into the study as control group. The bleeding index(BI),periodontal probing depth(PD)and clinical attachment loss(CAL) were recorded at 1 week after supragingival scaling, one month after subgingival scaling. Gingival crevicular fluid (GCF) was collected at 1 week after supragingival scaling, one week after subgingival scaling, one month after subgingival scaling. The level of SFRP1 in GCF samples was detected by ELISA. SPSS19.0 software package was used for data processing. RESULTS: The amount of SFRP1 in GCF of moderate to severe periodontitis group was (40.80±4.85) pg, and that of normal control was (33.42±2.24) pg at 1 week after supragingival scaling. The amount of SFRP1 in GCF was significantly higher in moderate to severe periodontitis compared to normal control group (P<0.05). In moderate to severe periodontitis group, the amount of SFRP1 in GCF significantly increased at l week after subgingival scaling (45.99±5.23) pg compared to 1 week after supragingival scaling and 1 month after subgingival scaling (36.92±4.00) pg (P<0.05); There was significant decrease in the amount of SFRP1 in GCF at 1 month after subgingival scaling,compared to 1 week after supragingival scaling and l week after subgingival scaling (P<0.05). CONCLUSIONS: The BI, PD in the periodontitis groups were significantly improved at 1 month after initial therapy,and the amount of SFRP1 in GCF changed with different periodontal inflammation state．

Comparative evaluation of cytokines in gingival crevicular fluid and saliva of patients with aggressive periodontitis.

OBJECTIVES: This study aims to evaluate and compare cytokines in gingival crevicular fluid (GCF) and saliva of patients with aggressive periodontitis (AP) before and after treatment. METHODS: Forty AP patients and 40 healthy volunteers were enrolled in this study. Clinical parameters included probing depth and sulcus bleeding index. GCF and saliva were collected from both groups. The levels of IL-1ß, IL-2, IL-4, IL-6, IFN-γ and TNF-α were measured using ELISA. RESULTS: The probing depth in AP patients was significantly deeper before treatment than after treatment. The concentrations of cytokines in GCF and saliva were significantly higher in AP patients than in the control group and decreased after periodontal treatment. Positive relationships were found between cytokine levels in GCF and clinical parameters. The reliability of cytokines in GCF and saliva was assessed by Cronbach's alpha analysis, which could be considered satisfactory. CONCLUSION: Cytokine levels in GCF and saliva correlated well with clinical parameters and AP. Measurements of cytokines in saliva may be regarded as a noninvasive and quick method for monitoring periodontal disease activity.

[Exploring the relationship between secreted frizzled-related protein 1 and chronic periodontitis].

OBJECTIVE: This study detects the expression of secreted frizzled-related protein 1 (SFRP1) in healthy patients and patients with chronic periodontitis (CP) and explores the relationship between SFRP1 and the occurrence and development of CP. METHODS: First, 28 patients forming the CP group were further divided into mild, moderate, and severe CP subgroups according to clinical attachment loss (CAL) data. Ten healthy volunteers were recruited in the control group. Gingival crevicular fluid (GCF) was collected from all of the patients, and the concentration of SFRP1 in the GCF samples was detected using enzyme-linked immunosorbent assay. Next, gingival lesions were obtained from 22 patients in the CP group and healthy gingival tissues were obtained from the 10 healthy patients in the control group. Immunohistochemical analysis for SFRP1 was used to analyze the correlation between the expression of SFRP1 and the severity of CP based on staining intensities. RESULTS: The concentration of SFRP1 in GCF samples taken from of the CP group (281.07 ng x L(-1) +/- 33.37 ng x L(-1)) was significantly higher than that in samples taken from the control group (245.30 ng x L(-1) +/- 35.69 ng x L(-1)) (P < 0.05). A significant positive correlation was observed between the concentration of SFRP1 in GCF and CAL (r = 0.651, P < 0.001). Furthermore, the SFRP1 scores in the CP groups (4.500 +/- 0.913) were significantly higher than those in the control group (2.800 +/- 1.135) (P < 0.001). SFRP1 scores did not vary significantly among the CP subgroups (P > 0.05). CONCLUSION: SFRP1 expression in the CP groups was significantly higher than that in the control group. Thus, SFRP1 may play a significant role in the development of CP.

Correlation of epigenetic change and identification of risk factors for oral submucous fibrosis.

BACKGROUND: DNA methylation of certain genes is an epigenetic change that is essential for tumorigenesis. Oral submucous fibrosis (OSF) is a precancerous condition of oral mucosa with inflammation and progressive fibrosis of the lamina propria and deeper connective tissue. The hypermethylation of E-cadherin and cyclooxygenase 2 (COX-2) in chronic inflammation may demonstrate a mild lesion/mutation at epigenetic levels. This study compares the hypermethylation status of E-cadherin and COX-2 genes in patients with oral cancer and patients with OSF and also aims to identify risk factors for the development of OSF. METHODS: DNA was extracted from blood samples of 50 healthy subjects, 50 patients with OSF and 60 patients with oral cancer. Methylation-specific polymerase chain reaction for E-cadherin and COX-2 was performed on these samples and the products were analyzed on 2% agarose gel. Surveys about oral health habits and clinical periodontal examinations in patients with OSF and healthy subjects were also conducted by well-trained dentists, and logistic regression was performed to identify risk factors for OSF. RESULTS: Hypermethylation of E-cadherin and COX-2 was observed in 36% and 22% of oral cancer samples, respectively. In patients with OSF, the rates were 52% and 30%, and in healthy controls the rates were 4% and 6%. Hypermethylation was shown to be correlated between the 3 groups with statistical significance (p<0.01). Methylation of CpG islands in E-cadherin and COX-2 occurred more frequently in patients with OSF than in the control group, but less frequently than in patients with oral cancer. In the logistic regression analysis, smoking, brushing more than twice daily, periodontal probing depth and plaque index were identified as 4 major risk factors for OSF. CONCLUSIONS: These data confirm that E-cadherin and COX-2 expressions are related to OSF. The epigenetic changes presented in patients with chronic inflammation might demonstrate an irreversible destruction in the tissues or organs similar to the effects of cancer. Chronic OSF was significantly associated with hypermethylation, a cancer risk factor.

Molecular property-affinity relationship of the interaction between dietary polyphenols and bovine milk proteins.

The relationship between the molecular properties of dietary polyphenols and their affinities for bovine milk proteins (BMP) was investigated. The affinities of polyphenols for BMP were determined by means of fluorescence titration. The affinities of polyphenols for BMP increased with increasing partition coefficient and decreased with increasing hydrogen bond acceptor number of the polyphenol. From this point, the hydrophobic force played an important role in the binding interaction between polyphenols. It was found that the topological polar surface area value decreases with increasing binding constant of the polyphenol for BMP, which illustrates that the glycosylation of hydroxyl groups in polyphenols weakens their binding affinity for BMP. A strong correlation between Mulliken electronegativity and binding affinity was found (R = 0.64626), and Mulliken electronegativity values were found to increase with increasing binding constant of polyphenols for BMP. This illustrates that electrostatic interactions play a key role in binding dietary polyphenols to BMP.

[Relationship between the chronic periodontitis and the depression anxiety psychological factor].

OBJECTIVE: To explore the relationship between the chronic periodontitis (CP) and the depression-anxiety psychological factors. METHODS: Thirty-one patients and 29 age, gender-matched volunteers were enrolled for this study. In order to assess the depression-anxiety psychological index, the subjects filled the questionnaire regarding the demographic and socioeconomic information, the oral hygiene habit, the Center for Epidemiologic Studies Depression Scale (CES-D) and the Self Rating Anxiety Scale(ASA). Calculus index (CI), bleeding on probing (BOP), probing depth (PD), clinical attachment loss (CAL), furcation involvement (FI) and tooth mobility were assessed at 6 sites per tooth of all erupted teeth by a manual periodontal probe. The data were analyzed by the analysis of variance, χ(2) test, and multivariable logistic step wise analysis via the software of SPSS 15.0. RESULTS: The mean CAL of the control group was 0.46 ± 0.16,the mean CAL of the moderate, high, and severe CP group was 2.84 ± 0.12, 3.51 ± 0.34, and 4.71 ± 0.51, respectively, which is significant difference between each other (P<0.01). The depression index of the volunteers, the moderate CP, the high CP, and the severe CP was 30.52 ± 3.73, 35.83 ± 7.76, 37.25 ± 6.16, 37.82 ± 5.94, respectively. The anxiety index among the 4 groups was 26.69 ± 3.55, 37.67 ± 6.31, 32.87 ± 5.54, and 35.94 ± 6.30, respectively. The depression and anxiety indexes of the periodontitis groups were higher than those of the control (P<0.01) while there was no significant difference among the 3 CP groups (P>0.05). The multivariate logistic analysis of the relationship between CP and the depression-anxiety psychological factors showed that the depression psychological factor was B=2.301,OR=9.988 while the optimistic coping style was B=-5.174,OR=0.006 in the equation of the regression. CONCLUSION: The depression psychological factor was related to the progression of CP. In addition, the optimistic coping style could prevent the progression of the CP.

Over-expression of CKS1B activates both MEK/ERK and JAK/STAT3 signaling pathways and promotes myeloma cell drug-resistance.

Here we demonstrate the crucial role of CKS1B in multiple myeloma (MM) progression and define CKS1B-mediated SKP2/p27(Kip1)-independent down-stream signaling pathways. Forced-expression of CKS1B in MM cells increased cell multidrug-resistance. CKS1B activates STAT3 and MEK/ERK pathways. In contrast, SKP2 knockdown or p27(Kip1) over-expression resulted in activation of the STAT3 and MEK/ERK pathways. Further investigations showed that BCL2 is a downstream target of MEK/ERK signaling. Stimulation of STAT3 and MEK/ERK signaling pathways partially abrogated CKS1B knockdown induced MM cell death and growth inhibition. Targeting STAT3 and MEK/ERK signaling pathways by specific inhibitors induced significant MM cell death and growth inhibition in CKS1B-overexpressing MM cells and their combinations resulted in synergy. Thus, our findings provide a rationale for targeting STAT3 and MEK/ERK/BCL2 signaling in aggressive CKS1B-overexpressing MM.

[Therapeutic effect of the combination of traditional Chinese medicine and western medicine on patients with oral submucous fibrosis].

OBJECTIVE: To observe the therapeutic effect of salvia miltiorrhiza and prednisolone on patients with oral submucous fibrosis (OSF). METHODS: A total of 60 medium-term OSF patients and 60 advanced stage OSF patients were randomly divided into the first group (treated with both salvia miltiorrhiza and prednisolone) and the second group (treated with prednisolone alone). The clinical effect was compared between each group after 3-month treatment. RESULTS: Difference was found in the lesion area of the medium-term cases and the advanced stage cases of the first group before the treatment [(10.37+/-3.40) cm2, (19.60+/-3.27) cm2] and after the treatment [(5.90+/-4.10) cm2, (16.33+/-4.02) cm2] (P<0.05). Significant difference was found in the mouth opening before the treatment [(3.41+/-0.77) cm, (1.98+/-0.39) cm] and after the treatment [(3.87+/-0.67) cm, (2.26+/-0.46) cm] (P<0.05) in the first group. There was significant difference in the lesion area and mouth opening of the medium-term cases of the second group before the treatment [(10.87+/-3.18) cm2, (3.57+/-0.75) cm] and after the treatment [(6.70+/-3.75) cm2, (3.97+/-0.69) cm] (P<0.05). No difference in the lesion area and mouth opening of the advanced stage cases of the second group was found (P>0.05). There was difference in the therapeutic efficacy between the first group (70%) and the second group (16.67%) of the advanced stage cases (P<0.05), but not in the clinical effect between the 2 groups of the medium-term cases (P>0.05). The side effect of prednisolone could be reduced while used together with salvia miltiorrhiza. CONCLUSION: There is obvious advantage in treating OSF by the combination of salvia miltiorrhiza and prednisolone.

[Periodontal status in patients with oral submucous fibrosis].

OBJECTIVE: To investigate the periodontal status in patients with oral submucous fibrosis (OSF), and to provide reference for the treatment and prophylaxis in patients with OSF and betel chewers. METHODS: Fifty samples clinically and pathologically diagnosed as OSF patients were selected as the OSF group, another 50 age-matched healthy volunteers in the similar living condition were compared with the OSF patients and non-betel nut chewers were classified as the control group. The 5 periodontal clinical parameters were collected and recorded, including plaque index, periodontal probing depth, clinical attachment loss, gingival index, and tooth count of bleeding of probing. RESULTS: There was a significant difference in plaque index (PLI) between the OSF group (2.14+/-0.64) and the control group (1.7+/-0.89) (P<0.01). Periodontal probing depth (PD) was (1.98+/-0.70) mm in the control group, and (5.57+/-2.39) mm in the OSF group, with significant difference in PD (P<0.01). There was no significant difference in clinical attachment loss, gingival index, and tooth count of bleeding on probing between the 2 groups (P>0.05). CONCLUSION: OSF patients tend to accumulate plaque, and have deep periodontal pocket, periodontal inflammation or severe periodontal damage.

[Expression of loricrin and cytochrome P450 3A5 in oral submucous fibrosis and their significance].

OBJECTIVE: To investigate the expression of loricrin (LOR) and cytochrome P450 3A5 (CYP 3A5) in oral submucous fibrosis (OSF) and to evaluate their roles in the defending ability of epithelium mucosae. METHODS: The expression of LOR and CYP 3A5 was examined in the specimens of 66 OSF and 14 normal buccal mucosa samples by immunohistochemistry, and the protein and mRNA expression of them was detected by Western blot and reverse transcriptase-PCR (RT-PCR). RESULTS: LOR was overexpressed in 42 (63.6%) cases of OSF, and showed a significant difference only between the early and moderately stages of OSF (P < 0.05), but no clear difference between moderately and advanced stages (P > 0.05). All normal buccal mucosa tissues showed positive immunoreactivity for CYP 3A5 protein in the membrane and cytoplasm of spinous epithelial cells and cytoplasm of endothelial cells, 5 (7.6%) cases of OSF showed weak staining of CYP 3A5 in spinous epithelial cells and 33 (50%) showed faint in cytoplasm of endothelial cells. A negative relationship between its expression and pathological stages was found in OSF (P < 0.05). RT-PCR results were fully consistent with the immunohistochemical data. But the results of Western blot only showed the expression of CYP 3A5 was significantly higher in normal buccal mucosa samples than OSF. CONCLUSION: The results suggest that the LOR and CYP 3A5 might play a vital role in the change of defending ability of epithelium mucosae as well as the pathopoiesis and carcinogenesis of OSF.

The phosphorylation of survivin Thr34 by p34cdc2 in carcinogenesis of oral submucous fibrosis.

Survivin is a crucial node molecule involved in apoptosis, cell division and drug discovery. Up-regulation of survivin in the tissues of oral submucous fibrosis (OSF) and oral squamous cell carcinoma (OSCC) originated from OSF has already been demonstrated. Survivin Thr34 phosphorylation is involved in the inhibition of apoptosis and cell division. To determine the potential involvement of survivin Thr34 phosphorylation in carcinogenesis of OSF, 40 OSFs, 42 OSCCs originated from OSF and 10 normal tissues from surgical specimens were studied. Immunohistochemistry showed that the positive staining rate of the survivin phosphorylation on Thr34 in OSCC originated from OSF group was significantly higher than that in OSF group (P<0.01), and none in the normal oral mucosa specimens. Survivin phosphorylation on Thr34 is predominantly located in the nucleus, which account for its function in apoptosis at cell division. Western blotting analysis showed increasing expression of survivin Thr34 phosphorylation, cyclin B1 and p34cdc2 in carcinogenesis of OSF. Furthermore, p34cdc2-cyclin B1 kinase was confirmed to phosphorylate survivin on Thr34 in carcinogenesis of OSF by immunoprecipitation and immunoblot. These results suggest that the phosphorylation of survivin on Thr34 critically regulate survivin and plays an important role during the malignant transformation of OSF, which will provide an indication to early diagnosis and therapy in carcinogenesis of OSF.

Discovery of novel biomarkers in oral submucous fibrosis by microarray analysis.

Oral submucous fibrosis (OSF) is a high-risk precancerous condition of the oral cavity. Areca nut chewing is its key etiologic factor, but the full pathogenesis is still obscure. In this study, microarray analysis was used to characterize the mRNA changes of 14,500 genes in four OSF and four normal buccal mucosa samples to identify novel biomarkers of OSF. Five candidate genes with the most differential changes were chosen for validation. The correlation between clinicopathologic variables of 66 OSF patients and the expression of each gene was assessed by immunohistochemistry. The microarray analysis showed that 661 genes were up-regulated (fold value >2) and 129 genes were down-regulated (fold value <0.5) in OSF (q < 0.01). The top three up-regulated genes [Loricrin, Cartilage oligomeric matrix protein (COMP), Cys-X-Cys ligand 9 (CXCL9)] with the largest fold changes and the top two down-regulated genes [keratin 19 (KRT19), cytochrome P450 3A5 (CYP 3A5)] with the most significantly differential changes in OSF were chosen as candidate biomarkers. In immunohistochemical results, the expression of Loricrin and COMP showed statistically significant association with histologic grade of OSF (P = 0.03 and 0.006, respectively). COMP was found to be overexpressed frequently in patients with the habit of areca nut chewing for more than 4 years (P = 0.002). CYP 3A5 was revealed an inverse correlation with histologic grade (P = 0.04). This pilot study showed that five novel genes might play important roles in the pathogenesis of OSF and may be clinically useful for early detection of OSF.