Activation Toll-Like Receptor7 (TLR7) Responsiveness Associated with Mitogen- Activated Protein Kinase (MAPK) Activation in HIOEC Cell Line of Oral Squamous Cell Carcinoma.

STATEMENT OF THE PROBLEM: Oral squamous cell carcinoma is the most common oral malignancy. Toll-like receptor (TLR) activation led to alterations in the levels of mRNA encoding the TLR accountable for recognizing the inducing agonist and cross-regulation of other TLR. PURPOSE: The purpose of this study is determination of mitogen-associated protein kinase (MAPK) activation in human immortalized oral epithelial cell (HIOEC) line via up regulating of TLR7. MATERIALS AND METHOD: expression of TLR7 was measured in HIOEC and normal cells by quantitative real-time polymerase chain reaction (qRT-PCR) and samples were calibrated by ß-actin. RESULTS: Western blot analysis discovered high expression of TLR7 and MAPK in HIOEC cell lines. TLR7 was over-expressed in HIOEC cell line. Imiquimod-induced expression of interleukin (IL)-6, IL-8, and vascular endothelial growth factor (VEGF) was inhibited by TLR7 siRNA in HIOEC cells as determined by reverse transcription polymerase chain reaction (RT-PCR). Mean fluorescence intensity of nuclear p38 expression was determined in HIOEC cell lines (p< 0.05). RT-PCR analysis of IL-6, IL-8, and VEGF mRNA expression in HIOEC cells stimulated with imiquimod (1 µg/ml) for indicated time points. CONCLUSION: TLR7 is functionally over-expressed in HIOEC cell line of oral squamous cell carcinoma and development of resistance to cisplatin in human oral squamous cell carcinoma might occur through the mechanism involving activation of TLR7 and its dependent signaling pathway.

Quercetin Decreases Th17 Production by Down-Regulation of MAPK- TLR4 Signaling Pathway on T Cells in Dental Pulpitis.

STATEMENT OF THE PROBLEM: Quercetin is a pharmacological flavonoid that can inhibit high mobility group box1 (HMGB1) protein, a non-histone nuclear protein that is implicated in inflammation. Th17 cells are important cells in the pathogenesis of inflammation. Pulpitis is the inflammation of dental pulp, which usually is accompanied by pain. Quercetin may alleviate this inflammation. PURPOSE: The current study aimed to compare blocking of HMGB1 function and stimulation of HMGB1 function with quercetin and investigate the effects of the blockage on T helper 17 (Th17) cells and mitogen-activated protein kinase Toll-like receptor 4 (MAPK-TLR4) signaling pathway. MATERIALS AND METHOD: T cells isolated from the pulp involved with pulpitis and the normal pulp were cultured. The cells suspensions were plated in 6-wells culture plates and stimulated with 0.5 µg/ml of HMGB1 for 2, 4, 8, and 12 hours. For blocking TLR4, 10 µg/ml rabbit anti-human TLR4 antibody was added 1 hour before treatment with HMGB1. RESULTS: The level of these cytokines decreased; moreover, western blot data showed that quercetin could decrease MAPK signaling pathway by means of inhibition of HMGB1 on T cells. The results showed the reduction of TLR4 pathway and Th17 cell polarization. CONCLUSION: Our results indicated that the levels of IL-17, IL-33, and IL-6 in supernatants from patients' cultured T cells were increased after stimulation with HMGB-1 following employing quercetin. It also could inhibit MAPK signaling pathway, which subsequently could decrease Th17 production and IL-17. Quercetin could decrease pro-inflammatory cytokines and IL-17 production.

Correlation between optic nerve involvement and chronic obstructive pulmonary disease.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a major public health problem worldwide. The aim of this study was to evaluate the rate of optic neuropathy in COPD patients. METHODS: Forty patients diagnosed with COPD and 60 healthy subjects as control group enrolled. After examination by a pulmonary subspecialist, patients were ranked by Global initiative for chronic Obstructive Lung Disease (GOLD) criteria, and patients with zero grades on GOLD criteria were excluded. Visual evoked potential by checkerboard (raster background) method with a frequency of 2 Hz were done for all participants. P-values less than 0.05 were considered as significant. RESULTS: Fifty-five percent of COPD patients had visual evoked potential abnormalities. Mean P100 latency in both eyes was significantly longer in COPD patients. Average P100/N140 amplitude in both eyes were insignificantly higher in COPD. CONCLUSION: Higher P100 latency in COPD patients shows demyelinating type of optic nerve involvement; however, further investigation in this area is needed.

The effect of interleukin 36 gene therapy in the regression of tumor.

BACKGROUND: Cancer immunotherapy attempts to stimulate the immune system to reject and destroy tumors and is one of the cancer treatment strategies. Recently, interluekin36 (IL36) has been used as immunotherapeutic agents in cancer gene therapy. Present study investigated that the IL36 gene therapy effects on the regression of tumor masses in mouse model. Aim of this study is determination of the gene therapy effects by IL36 in the regression of tumor masses in mouse model. METHODS: To study the therapeutic efficacy of this cytokine, WEHI-164 tumor cells were transected with mIL36 plasmids. ELISA test was used to check cytokine production by transected cells. To establish fibro sarcoma mouse model, Tumoral transfected cells were injected subcutaneously to inoculate tumor in BALB/C mice. Tumor volumes were measured by caliper. Mice were sacrificed and tumors were extracted. The expression of IL36 and IFN-γ was studied with Real-time PCR and immunoblotting. The expression of Ki-67 (a tumor proliferation marker) in tumor masses was studied by immunohistochemistry staining. In this study we had 2 groups which are treated with IL-36 and Untreated with IL-36 as a blank. RESULTS: The group treated with IL36 indicated decrease of tumor mass volume (p<0.001). The results of western blotting and real-time PCR showed the IL36 expression increased in the group treated with IL36 (with relative expression of 1.9). CONCLUSION: Immunohistochemistry staining indicated that the Ki-67expression has been reduced in the group interfered with IL36. IL36 gene therapy has therapeutic effects on the regression of tumor masses in fibro sarcoma mouse model.

Application of crestal anesthesia for treatment of class I caries in posterior mandibular teeth.

BACKGROUND AND AIMS: Current infiltration techniques for achieving anesthesia in dental procedures are not applicable in posterior mandibular region because of its dense cortical bone. The aim of this study was to evaluate the efficacy of a specific infiltration anesthesia in posterior mandibular teeth instead of inferior alveolar nerve block for restorative procedures. MATERIALS AND METHODS: Crestal anesthesia (CA) was assessed both clinically and by computed tomography scan for its efficacy and side effects. A combination of an opaque material (Ultravist) and 2% lidocaine was used to trace the anesthetic solution. The combination was primarily injected in the gingival tissue of rabbit and was followed-up regularly for two weeks to assess any possible injury. After confirming its safety, a combination of these materials was injected to volunteers to assess efficacy and diffusion route. A total of 154 patients (77 female, 77 male) with matched bilateral posterior teeth in mandible were selected randomly and an IANB and CA were performed randomly and separately in different sessions for the contra lateral teeth. The onset of anesthesia, anesthesia duration, pain, blood pressure, pulse rate, and consumed volume of anesthetic solution was recorded for each technique. Data were analyzed using paired t-test. RESULTS: There were no significant differences in clinical attachment loss, pocket depth, bone level, plaque index, and free gingival margin between the two flaps (p>0.05). CONCLUSION: CA could be considered as a reliable and safe primary injection in posterior mandibular teeth for restorative treatments.