Case report: postradiation chondrosarcoma with a short latency period of 6 months.

We describe a case of postradiation chondrosarcoma after basal cell carcinoma treatment. At the time he presented, the patient was a 35-year-old man who had received radiotherapy at a dose of 70 Gy for 8 weeks. Six months after radiation treatment, a rapidly growing mass at the upper right alveolar ridge of the gums, where radiation had been given, was diagnosed as chondrosarcoma. Generally, chondrosarcoma occurs after a latency period of several years following radiation. However, there are a few relevant reports indicating that maxillofacial chondrosarcoma can develop after radiotherapy for basal cell carcinoma, with a short latency of 6 months. We hypothesize that the dosage and treatment time of radiation may have played a role in the opening/closing of the Hh-signaling pathway in the case of this patient.

Epigenetic changes of TIMP-3, GSTP-1 and 14-3-3 sigma genes as indication of status of chronic inflammation and cancer.

OBJECTIVES: This study aimed to compare the epigenetic changes via hypermethylation status of TIMP-3, GSTP-1 and 14-3-3σ genes, between healthy subjects and patients with reversible chronic inflammatory disease, and between healthy subjects and patients with irreversible malignant disease, to highlight the genetic changes that occur in the progression from an inflammatory condition to irreversible genetic changes commonly observed in cancer patients. METHODS: DNA was extracted from the blood of 680 healthy subjects, and tissues and blood of 110 patients with chronic inflammation disease of the gums, as well as neoplastic tissues of 108 breast cancer patients. Methylation-specific polymerase chain reaction (PCR) for TIMP-3, GSTP-1 and 14-3-3σ was performed, and hypermethylation status was analyzed and compared between the 3 groups. RESULTS: The hypermethylation frequencies of TIMP-3 and GSTP-1 of reversible chronic inflammatory gum disease and the control group were similar, but both were significantly lower than those for malignant disease patients (p<0.0001). The methylation frequency of 14-3-3σ in chronic inflammatory gum disease was higher than in the cancer and control groups (p<0.0001). The methylation of CpG islands in TIMP-3 and GSTP-1 in chronic inflammation patients occurred as frequently as in the control group, but less frequently than in breast cancer patients. However, the epigenetic silencing of 14-3-3σ occurred more frequently in the chronic inflammation group than in cancer patients and healthy controls. CONCLUSIONS: The epigenetic silencing of 14-3-3σ might be essential for chronic inflammatory gum disease. The epigenetic changes presented in chronic inflammation patients might demonstrate an irreversible destruction in the tissues or organs similar to cancer.

Angiotensin II induces mitochondrial dysfunction and promotes apoptosis via JNK signalling pathway in primary mouse calvaria osteoblast.

OBJECTIVES: This present study was designed to investigate the effects of Angiotensin II on mitochondrial functions, ROS generation and c-jun N-terminal kinases (JNK) signalling pathway-mediated cell apoptosis in mouse calvaria osteoblasts. METHODS: Calvaria osteoblast were isolated and cultured. The cells were separated into two groups-control and treated groups-where the latter was stimulated with angiotensin II (Ang II). Mitochondrial reactive oxygen species (ROS) and superoxide production were measured. Intracellular ATP levels were also detected. The cell proliferation rate was determined for the two groups. Protein production such as Anti-Bax, Bcl-2, COX IV and activation of c-jun N-terminal kinases signal (JNK) pathway was measured by enzyme-linked immunosorbent assay (ELISA) methods and Western blotting in this study. RESULTS: Ang II treated cells showed significantly higher levels of superoxide production compared to the control group (p<0.05). Conversely, Ang II induced inhibitory effects on mitochondrial respiratory enzyme complexes, cause membrane potential dissipation, ATP loss and promote ROS generation, cell apoptosis in cultured osteoblasts. In addition, JNK phosphorylations were involved in activating the mitochondria-dependent apoptotic pathway following Ang II stimulation, as pre-treatment of JNK-specific inhibitor SP600125 could rescue osteoblast cells from apoptosis by enhancing the anti-apoptotic protein Bcl-2 expressions, suppressing the translocation of Bax from cytosol into mitochondria, blocking cytochrome C release and caspase-3 activation. CONCLUSIONS: Ang II stimulates osteoblast apoptosis via suppression of the mitochondrial respiratory enzymes, membrane potential and cellular ATP productions. Clinical application with Ang II-stimulated osteoblast could be used for modelling or bone resorption in the oral region.

Status of oral ulcerative mucositis and biomarkers to monitor posttraumatic stress disorder effects in breast cancer patients.

BACKGROUND: This study was designed to assess oral ulcerative mucositis, C-reactive protein, blood pressure, heart rate and thyroid function in breast cancer patients in relation to the occurrence of posttraumatic stress disorder (PTSD). METHODS: A total of 120 female breast cancer patients and women 100 healthy subjects were enrolled in this study. PTSD status was assessed by questionnaire. Before and after treatment (modified radical mastectomy and chemotherapy), serum samples were collected and measured for levels of triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH) and high-sensitivity C-reactive protein (hs-CRP) by ELISA. Oral ulcerative mucositis was evaluated by the number and duration of oral ulcers and the degree of pain. RESULTS: Breast cancer patients experienced long-term PTSD and had elevated serum T3 and T4 levels. Patients experienced more severe pain and longer duration of oral ulcers compared with the healthy group. Oral ulcers were significantly associated with PTSD score in terms of the number of ulcers (p=0.0025), the degree of pain (p<0.0001) and the duration of ulcers (p<0.0001). CONCLUSION: These findings support that thyroid function is altered in breast cancer patients with PTSD. Elevation of T3 and T4 and oral ulcerative mucositis might be indicative of the emotional status of breast cancer patients.

Association of cytokines, high sensitive C-reactive protein, VEGF and beta-defensin-1 gene polymorphisms and their protein expressions with chronic periodontitis in the Chinese population.

PURPOSES: This study aimed at investigating the association between interleukin-6 (IL-6), interleukin-12 (IL-12), C-reactive protein (CRP), vascular endothelial growth factor (VEGF) and ß-defensin-1 polymorphisms and the susceptibility to periodontitis in the Chinese population. METHODS: DNA was extracted from the blood samples of 532 healthy individuals and 122 chronic periodontitis (CP) patients enrolled in the study. The genes encoding IL-6, IL-12, CRP, VEGF and ß-defensin-1 were amplified using PCR and digested with restriction enzymes. The protein expression of the abovementioned genes was determined by ELISA. Differences in the allele/genotype frequencies were assessed with the chi-square test. RESULTS: The frequencies of the C/C genotypes of IL-6, IL-12, and VEGF were higher in CP patients than healthy controls (66.3% vs 25.9%; 27.8% vs 19.9%; and 64.8% vs 52.1%, respectively). In the patients' group we also recorded frequencies of the A/A genotypes of CRP and VEGF higher than in healthy controls (63.1% vs 58.1% and 64.8% vs 35.2%, respectively). Protein production evaluated by ELISA demonstrated significant differences between CP patients and healthy controls for IL-6, IL-12, CRP, VEGF and ß-defensin-1. CONCLUSIONS: The genotypes of IL-6, IL-12, VEGF and ß-defensin-1 and their protein productions were associated with CP in a Chinese population. Genotypes and serum levels of CRP were associated with CP, but alleles frequency showed no difference between CP patients and healthy controls.

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.

Application of interleukin-1 genes and proteins to monitor the status of chronic periodontitis.

It has been widely reported that periodontitis may lead to bone tissue and teeth loss and result in failure of prosthodontics or implants. Interleukin-1 (IL-1) is a potent proinflammatory cytokine that plays an essential role during the pathogenesis of periodontitis. However, the gene polymorphisms of IL-1α, IL-1ß and IL-1RN and the relationship between these protein expressions in healthy people and patients with chronic periodontitis (CP) in China have not been fully elucidated. We investigated the gene polymorphisms and protein expression of IL-1α, IL-1ß and IL-1RN in healthy subjects and CP patients, and our data suggest that these gene polymorphisms are associated with CP. The frequency of the C/C genotype of IL-1α was 55% in CP patients, while in the control group it was 20% (p<0.0001). The C/C genotype of IL-1ß was also higher in CP patients (51%) than in controls (21%) (p<0.0001). For the 2/2 genotype of IL-1RN, CP patients showed a 30% frequency, while in controls this was 15% (p<0.0001). Protein levels evaluated by enzyme-linked immunosorbent assay demonstrated a significant difference in secretion between patients and controls for IL-1α and IL-1ß. These results indicate that genotype and protein production of IL-1α, IL-1ß and IL-1RN are associated with CP in a Chinese population, and might be putative risk indicators for chronic periodontitis.

Evaluation of the psychological and biological changes of patients diagnosed with benign and malignant breast tumors.

PURPOSE: Posttraumatic stress disorder (PTSD) is a severe anxiety disorder developed by exposure to any incident or circumstance that results in psychological trauma. In this study we compared the psychological and physiological changes between patients with malignant and benign breast tumors. METHODS: We selected 150 Chinese women with a breast mass, aged 20 to 45 years, from the Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital between 2009 and 2011 for this study; 30 healthy participants were enrolled into the control group. All subjects were examined and had their tumor mass aspirated for diagnosis. Equal numbers of patients with benign and malignant tumors were recruited. Patients with malignant tumors presented with low grade, minimal tumor invasion and non-involved lymph nodes. Questionnaires regarding anxiety, depression and PTSD were conducted 2 hours before getting the diagnostic result and 1 month after the diagnosis. Serum levels of IL-6, TNF-α, cortisol and high-sensitivity C-reactive protein before and after diagnosis were investigated and compared. The number of occurrences of oral ulcerative mucositis was also recorded. RESULTS: All patients experienced a certain degree of anxiety and their biomarkers were elevated compared with the normal reference range before the pathological report was disclosed. However, 1 month after the operation, the benign tumor group showed significantly lower levels of biomarkers and anxiety scores than patients with a malignant breast tumor. The results were consistent throughout 12 months of study. CONCLUSION: Study subjects with a benign tumor returned to their normal condition after being diagnosed, while patients with a malignant tumor suffered from a certain degree of PTSD or depression.

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.

Application of a nanotechnology antimicrobial spray to prevent lower urinary tract infection: a multicenter urology trial.

BACKGROUND: Catheter-associated urinary tract infection (CAUTI) is a common nosocomial device-associated infection. It is now recognized that the high infection rates were caused by the formation of biofilm on the surface of the catheters that decreases the susceptibility to antibiotics and results in anti-microbial resistance.In this study, we performed an in vitro test to explore the mechanism of biofilm formation and subsequently conducted a multi-center clinical trial to investigate the efficacy of CAUTI prevention with the application of JUC, a nanotechnology antimicrobial spray. METHODS: Siliconized latex urinary catheters were cut into fragments and sterilized by autoclaving. The sterilized sample fragments were randomly divided into the therapy and control group, whereby they were sprayed with JUC and distilled water respectively and dried before use.The experimental standard strains of Escherichia coli (E. coli) were isolated from the urine samples of patients. At 16 hours and 7 days of incubation, the samples were extracted for confocal laser scanning microscopy.A total of 1,150 patients were accrued in the clinical study. Patients were randomized according to the order of surgical treatment. The odd array of patients was assigned as the therapy group (JUC), and the even array of patients was assigned as the control group (normal saline). RESULTS: After 16 hours of culture, bacterial biofilm formed on the surface of sample fragments from the control group. In the therapy group, no bacterial biofilm formation was observed on the sample fragments. No significant increase in bacterial colony count was observed in the therapy group after 7 days of incubation.On the 7th day of catheterization, urine samples were collected for bacterial culture before extubation. Significant difference was observed in the incidence of bacteriuria between the therapy group and control group (4.52% vs. 13.04%, p < 0.001). CONCLUSIONS: In this study, the effectiveness of JUC in preventing CAUTI in a hospital setting was demonstrated in both in vitro and clinical studies.

Comparing serum levels of cardiac biomarkers in cancer patients receiving chemotherapy and subjects with chronic periodontitis.

BACKGROUND: Chronic periodontitis (CP) is a chronic inflammation associated with elevations of several inflammatory and cardiac markers. Studies implicated CP as one of the etiologies in coronary heart disease (CHD). Cardiotoxicity is a major complication of anticancer drugs, including anthracyclines and 5-fluorouracil (5FU). The most severe cardiac complications are heart failure, arrhythmia and coronary heart disease (CHD). In this study, we compared the level of inflammatory factors and cardiac markers between chronic periodontitis patients and cancer patients receiving chemotherapy. METHODS: 108 blood samples of periodontally healthy subjects were obtained on random from Hong Kong Red Cross, and these represented the controlled population. Forty-four patients diagnosed with chronic periodontitis were recruited from the West China Hospital of Stomatology, Sichuan University. They have received scaling and root planning with mean pocket depths of 6.05 mm. Thirty breast cancer patients diagnosed with invasive ductal carcinoma from UNIMED Medical Institute, Hong Kong gave consent to participate in this study. They received 4 cycles of 500mg/m2 5-fluorouracil, 75 mg/m2 epirubicin and 500mg/m2 cyclophosphamide at a 3-week interval between each cycle. Peripheral venous blood from each group was taken for measurement of blood cells, inflammatory marker (P-selectin, high sensitvity C-reactive protein) and cardiac markers (troponin T; troponin I; N-terminal pro brain natriuretic peptide (Nt-proBNP) and Lactate dehydrogenase (LDH). RESULTS: The lymphocyte count was higher (p < 0.05) in periodontitis patients than the other two groups, and more neutrophils (p < 0.05) were seen in cancer patients receiving chemotherapy. The two test groups demonstrated higher levels (p < 0.01) of inflammatory and cardiac markers than the control group. CONCLUSIONS: The elevated cardiac markers found in periodontitis patients suggested that they may carry potential risks in developing cardiac lesions. Troponin T, troponin I, pro-BNP, LDH and high sensitvity C-reactive protein may be used as markers to monitor cardiac lesions in chronic inflammatory patients.

Gene polymorphism and protein of human pro- and anti-inflammatory cytokines in Chinese healthy subjects and chronic periodontitis patients.

BACKGROUND: Periodontal disease is thought to arise from the interaction of various factors, including the susceptibility of the host, the presence of pathogenic organisms, and the absence of beneficial species. The genetic factors may play a significant role in the risk of periodontal diseases. Cytokines initiate, mediate and control immune and inflammatory responses. The aim of this study is to compare genotypes and soluble protein of pro and anti-inflammatory cytokines (IL-1α, IL-1ß, IL-6, IFN-γ, IL-10, TNF-α and IL-4) in subjects with or free of chronic periodontitis. METHODS: A total of 1,290 Chinese subjects were recruited to this clinical trial: 850 periodontally healthy controls and 440 periodontal patients. All subjects were free of systemic diseases. Oral examinations were performed, and the following parameters were recorded for each subject: supragingival/subgingival calculus, gingival recession, bleeding on probing (BOP), probing depth (PD), clinical attachment loss (CAL), gingival recession and tooth mobility. The peripheral blood samples were collected for genetic and enzyme linked immunosorbent assay (ELISA) analysis. Restriction enzymes were used for digestion of amplified fragments of IL-1α, IL-1ß, IL-6, IFN-γ, IL-10, TNF-α and IL-4. RESULTS: The protein expressions of patient and control samples for IL-1α, IL-1ß, IL-6, TNF-α, IFN-γ, IL-10, and IL-4 measured by ELISA confirmed a statistically significant difference (p < 0.001). The digestion of fragments of various genes showed that the pro-inflammatory cytokines IL-1α and TNF-α, and the anti-inflammatory cytokines IL-4 and IL-10 demonstrated a correlation with chronic inflammation in patients (X2: p < 0.001). The remaining genes investigated in patients and healthy subjects (IL-1ß, IL-6, IFN-γ and IL-10) did not show any significant difference. CONCLUSIONS: The cytokine gene polymorphisms may be used as a marker for periodontitis susceptibility, clinical behaviour and severity. This detection offers early diagnosis and induction of prophylaxis to other family members against disease progression.

Clinical application of human ß-defensin and CD14 gene polymorphism in evaluating the status of chronic inflammation.

BACKGROUND: Periodontitis is a common disease that affects the periodontal tissue supporting the teeth. This disease is attributed to multiple risk factors, including diabetes, cigarette smoking, alcohol, pathogenic microorganisms, genetics and others. Human beta-defensin-1 (hBD-1) is a cationic antimicrobial peptide with cysteine-rich ß-sheets and broad-spectrum antimicrobial activity. CD14 is a protein involved in the detection of bacterial lipopolysaccharide (LPS) and has also been associated with periodontitis. This study investigates the single nucleotide polymorphic (SNP) region, -1654(V38I), of the human beta-defensin-1 (hBD-1) gene as well as the -159 region of the CD14 gene in subjects with chronic periodontitis. METHODS: Blood samples from periodontally healthy subjects and periodontitis patients were obtained. DNA was extracted from the blood and was used to perform restriction digest at the polymorphic G1654A site of DEFB1 with the enzyme HincII. The polymorphic site 159TT of CD14 was digested with the enzyme AvaII. Enzyme-linked immunosorbent assay (ELISA) was performed on soluble samples to determine the protein expressions. RESULTS: The control and patient groups expressed 35% and 38% 1654 A/A genotype of DEFB1, respectively. The A allele frequency of the control group was 40%, while the patient blood group was 54%. The mean hBD-1 protein levels of the control and patient samples were 102.83 pg/mL and 252.09 pg/mL, respectively. The genotype distribution of CD14 in healthy subjects was 16% for C/C, 26% for T/T and 58% for C/T. The genotype frequencies of CD14 in periodontitis patients were 10% for C/C, 43% for T/T and 47% for C/T. The CD14 protein expression determined by ELISA showed a mean protein level of the control samples at 76.28ng/mL and the patient blood samples at 179.27ng/mL with a p value of 0.001.Our study demonstrated that patients suffering from chronic periodontitis present more commonly with the 1654A/A genotype on the DEFB1 gene and the 159T/T genotype on the CD14 gene. CONCLUSIONS: This study purely investigated the association between periodontitis and one polymorphic site on both DEFB1 and CD14 gene, with the purpose of expanding knowledge for the future development in diagnostic markers or therapeutic interventions to combat this disease.

Association of matrix metalloproteinase (MMP)-1, 3, 9, interleukin (IL)-2, 8 and cyclooxygenase (COX)-2 gene polymorphisms with chronic periodontitis in a Chinese population.

Chronic periodontitis (CPs) could result in damage of periodontal tissues, loss of teeth and impose troublesome hindrance to restore teeth satisfyingly as well. Functional gene polymorphisms of matrix metalloproteinases, cytokines and cyclooxygenase-2 have been found to play important roles in periodontitis. This study was to investigate the association between MMP-1-1067, MMP-3-1171, MMP-9-1562, IL-2-330, IL-8-251, COX-2-765 polymorphisms, and the susceptibility to CP in a Chinese population. A total of 122 patients with CP were evaluated for MMP-1, MMP-3, MMP-9, IL-2, IL-8 and COX-2 genetic polymorphisms and were compared with 532 healthy control subjects using PCR-RFLP analysis. Clinical periodontal parameters were recorded. Serum levels of MMP-1, MMP-3, MMP-9, IL-2, IL-8 and COX-2 were measured by ELISA. The data were analyzed by chi-square, logistic regression and Mann-Whitney-U-tests and t test. There were significant differences between CP patients and healthy subjects in the genotype distribution and allele frequency of MMP-3-1171, MMP-9-1562, IL-2-330, IL-8-251 and COX-2-765 genetic polymorphisms. Significant difference between patients and controls were also observed for MMP-1-1067 genotype frequency, but not for allele frequency. Differences between rare allele carriage rates of CP and healthy groups regarding all the genetic polymorphisms in our study were significant (p<0.05). Serum levels of all the cytokines were higher in the CP patients compared to healthy subjects. These data show that MMP-1-1067, MMP-3-1171, MMP-9-1562 and IL-8-251 polymorphisms are associated with susceptibility to CP. MMP-1-1067 2G, MMP-3-1171 6A, MMP-9-1562 T and IL-8-251 A allele are associated with decreased susceptibility to CP in Chinese population.

Association of matrix metalloproteinase (MMP-1, MMP-3 and MMP-9) and cyclooxygenase-2 gene polymorphisms and their proteins with chronic periodontitis.

AIMS: The objective of this study was to investigate the association amongst the single nucleotide polymorphisms of genes encoding for matrix metalloproteinase (MMP) 1, 3, 9 and cyclooxygenase-2 (COX-2) of subjects. Protein production of MMPs, COX-2 and Vascular Endothelial Growth Factor (VEGF) were also investigated. METHODS: 280 chronic periodontitis patients and 250 periodontitis-free subjects were selected. DNA was extracted from blood samples of all patients, the polymorphic sites of the genes that encode for metalloproteinases and cyclooxygenase-2 were amplified using PCR, and digested with restriction enzymes. ELISA was used to determine the protein production of MMPs, COX-2 and VEGF. RESULTS: The mean probing depth (PD) was 5.4mm and the clinical attachment loss (CAL) was 6.4mm in patients group with at least 2 years history. 2G/2G genotype of MMP-1, the periodontitis patients presented frequency of 28% and the control only showed 3%. 5A/5A genotype of MMP-3, the periodontitis patients presented higher frequency of 55% than the control 40%. C/C of genotype MMP-9, the periodontitis patients presented higher frequency of 51% than the control 17%. C/C of genotype COX-2, the periodontitis patients demonstrated 28% frequency and the control was 3%. ELISA analysis determined a significant difference (p<0.001) in protein production between patient and control samples for the bio-markers. 12 cases with suspicious genotype of MMPs and in COX-2 showed the serum level was the highest value between other C/C genotype. CONCLUSIONS: Combine genotype and serum expression of inflammatory mediators that may be a good bio-marker for diagnosis and prognosis of the periodontitis.

Angiotensin II induces interleukin-6 synthesis in osteoblasts through ERK1/2 pathway via AT1 receptor.

BACKGROUND: Interleukin-6 (IL-6) is a potent stimulator of osteoclastic bone resorption. Osteoblast secretion of IL-6 plays an important role in the regulation of bone metabolism. Angiotensin II (Ang II) has been shown to regulate the expression of potent inflammatory factors, including MCP-1 and IL-6, by stimulating endothelial cells, vascular smooth muscle cells (VSMC) and monocytes. However, of the mechanism by which Ang II regulates IL-6 expression in osteoblasts is unknown. AIMS: The present study was designed to investigate the effect of Ang II on IL-6 expression in osteoblasts isolated from mice. The receptor(s) required and the potential role of extracellular signal-regulated kinase 1/2 (ERK1/2) activation in Ang II-induced IL-6 synthesis was also examined in these cells. METHODS: The osteoblasts were isolated from the calvaria of mice and cultured in α-MEM medium. IL-6 mRNA expression and protein synthesis was determined by qPCR and ELISA analyses. ERK1/2 kinase activation was determined by western blot. RESULTS: The results indicate that Ang II induced IL-6 mRNA expression and protein synthesis in cultured osteoblasts. However, these effects were abolished by pre-treatment with Ang II type 1 (AT1) receptor antagonist, losartan, and the ERK1/2 inhibitor, U0126, inhibited Ang II-mediated IL-6 expression and the phosphorylation of ERK1/2. CONCLUSION: Ang II induces the synthesis of IL-6 in osteoblasts through activation of the ERK1/2 pathway via the AT1 receptor.

Epigenetic change in E-cadherin and COX-2 to predict chronic periodontitis.

BACKGROUND: DNA methylation of certain genes frequently occurs in neoplastic cells. Although the cause remains unknown, many genes have been identified with such atypical methylation in neoplastic cells. The hypermethylation of E-Cadherin and Cyclooxygenase 2 (COX-2) in chronic inflammation such as chronic periodontitis may demonstrate mild lesion/mutation epigenetic level. This study compares the hypermethylation status of E-Cadherin and COX-2 genes which are often found in breast cancer patients with that in chronic periodontitis. METHODS: Total DNA was extracted from the blood samples of 108 systemically healthy non-periodontitis subjects, and the gingival tissues and blood samples of 110 chronic periodontitis patient as well as neoplastic tissues of 106 breast cancer patients. Methylation-specific PCR for E-Cadherin and COX-2 was performed on these samples and the PCR products were analyzed on 2% agarose gel. RESULTS: Hypermethylation of E-Cadherin and COX-2 was observed in 38% and 35% of the breast cancer samples, respectively. In chronic periodontitis patients the detection rate was 25% and 19% respectively, and none was found in the systemically healthy non-periodontitis control subjects. The hypermethylation status was shown to be correlated among the three groups with statistical significance (p < 0.0001). The methylation of CpG islands in E-Cadherin and COX-2 genes in periodontitis patients occurs more frequently in periodontitis patients than in the control subjects, but occurs less frequently than in the breast cancer patients. CONCLUSIONS: This set of data shows that the epigenetic change in E-Cadherin and Cyclooxygenase-2 is associated with chronic periodontitis. The epigenetic changes presented in chronic inflammation patients might demonstrate an irreversible destruction in the tissues or organs similar to the effects of cancer. Chronic periodontitis to some extent might be associated with DNA hypermethylation which is related to cancer risk factors.

Combination of radiological and biochemical methods to assess bone mineral density of mandible in fully edentulous patients after chemotherapy: a 5-year prospective study.

BACKGROUND: Osteoporosis is of concern in breast cancer patients who are undergoing chemotherapy. This study compared the bone mineral density (BMD) index of the mandible and hip hinge between patients who were undergoing chemotherapy or breast cancer, and fully edentulous Chinese patients without cancer over a period of 5 years. METHOD: 120 fully edentulous patients with an average age of 69 who had undergone mastectomy for grade two or three non-metastatic invasive breast ductal carcinoma. This was followed by administration of 5-fluorouracil, cyclophosphamide, and doxorubicin. The 118 fully edentulous cancer-free patients were included as a control group. The first assessment point was 6 months after chemotherapy treatment. The BMD and panoramic and side views of the mandible were measured by gamma-ray and the BMD of left hip hinge was measured using dual energy X-ray absorptiometry. The serum and urine level of bone-specific alkaline phosphatase (BAP), carboxyterminal cross-linked telopeptide of type I collagen (ICTP), as well as liver and renal function tests were determined. These examinations were performed annually for 5 years. RESULT: The cancer patients demonstrated a statistically significant (p < 0.05) increase in bone resorption of mandible and hip, and an increase in BAP and ICTP levels when compared with the control group. Although data were collected annually there was no statistical significance for the first 3 years. CONCLUSION: Breast cancer patients undergoing chemotherapy displayed significant resorption of mandibular bones compared with the healthy control, which might result in difficulties in fitting dentures, as it would cause pain and mucosal friction. Thus, concurrent therapy to decrease mandibular bone loss and special considerations in dentures are warranted for these patients.

Rhodiola algida improves chemotherapy-induced oral mucositis in breast cancer patients.

OBJECTIVE: Oral ulcerative mucositis, a common adverse effect due to mainstream cytotoxic drugs, limits the nutritional intake of cancer patients. Rhodiola algida is widely used in traditional Chinese medicine to stimulate the immune system. The aim of this study is to investigate the effect of this herbal extract on healthy human lymphocytes in vitro, the homeostasis of cancer patients and the healing time of oral ulcers. METHOD: The constituents of Rhodiola algida were analyzed by RP-HPLC. Lymphocytes isolated from 462 healthy subjects were treated with 100 ug/ml Rhodiola algida for 48 h. The activity of the cells was measured by cell proliferation reagent and ATP assay. The level of various cytokines and mRNA content of lymphocytes were determined. Rhodiola algida demonstrated no toxicity in animals, which had been orally fed with 1 mg/ml Rhodiola algida for 30 days. 130 breast cancer patients from Huaxi Hospital of Sichuan University were recruited between 2006 and 2007. They received four cycles of 5-fluorouracil, epirubicin and cyclophosphamide after modified total mastectomy. These patients were randomly assigned to test and control groups. Rhodiola algida mixture was consumed by the test group for 14 consecutive days after each cycle of chemotherapy. All patients were given 0.2% chlorohexidine mouth wash to be used every day. Complete blood counts, liver and renal function tests together with the number and size of oral ulcerations were analyzed after each cycle. Weight loss, complaints of nausea or vomiting and degree of pain were noted. RESULTS: The optimal concentration of Rhodiola algida favored the proliferation of lymphocytes. The levels of IL-2, IL-4, granulocyte-macrophage colony-stimulating factor and the mRNA content of these cytokines were also enhanced. White blood cell (WBC) levels returned to normal range in both groups 1 week after every cycle of chemotherapy. WBC count increased faster in patients using Rhodiola algida; they presented with smaller and fewer oral ulcers. There were no liver or renal complications observed in any patients. CONCLUSION: Rhodiola algida increases immunity of patients who are receiving chemotherapy post mastectomy and decreases the quantity of oral ulcers. Thus Rhodiola algida has the potential to be used concurrently with chemotherapy to alleviate the occurrence of oral ulcers.

Fructus schisandrae (Wuweizi)-containing compound inhibits secretion of HBsAg and HBeAg in hepatocellular carcinoma cell line.

Chronic hepatitis B is probably the major cause of cirrhosis and hepatocellular carcinoma. The detection of the hepatitis B virus surface antigen (HBsAg) and HBV core protein, the e antigen (HBeAg), indicates infection with the hepatitis B virus and replication activity, respectively. Fructus schisandrae containing compound (KY88) may affect the elimination of HBV, strengthen the immune system, as well as stimulate liver cell regeneration. The present study was conducted to demonstrate the ability of KY88 in inhibiting hepatocellular carcinoma cell proliferation and secretions of HBsAg and HBeAg. The hepatocellular carcinoma cell line HB-8064 was treated by KY88 followed by the measurement of cell proliferation rate and secretions of HBsAg and HBeAg on days 1, 3, 5, and 7. A semi-quantitative RT-PCR method was used to quantify the expression of the mRNA. Seventy Sprague-Dawley rats were fed for 28days with purified KY88 for a toxicity test. The expression of surface and e antigens was lower when the cells were treated for a longer time with KY88 or when the doses were higher. One-way ANOVA analysis confirmed the mRNA content of HBsAg to be significantly less than control. The body weight did not show a significant difference compared to the control group. Fructus schisandrae-containing compound (KY88) was potentially effective in suppressing the proliferation of hepatocellular carcinoma cells. The decreased secretion and gene expression of HBsAg and HBeAg might restrict the growth of tumour masses.