Cyclosporin A suppresses Porphyromonas gingivalis lipopolysaccharide induced matrix metalloproteinases activities in the co-culture of human gingival fibroblasts and monocyte cell line THP-1.

Cyclosporine-A (CsA) is a widely used immunosuppressant. In this study, we explore the pathway through which CsA suppressed the Porphyromonas gingivalis lipopolysaccharide (P.g-LPS)-induced increase in matrix metalloproteinase (MMP) activities in co-cultured human gingival fibroblasts (HGFs) and THP-1 monocytes. In the co-culture, we found that CsA inhibited the expression of cyclophilin A (CyPA), CD147 and the activities of MMPs, which were all induced by P.g-LPS. We also found that P.g-LPS and recombinant human CyPA increased activation of ERK1/2 and IκB (an NF-κB inhibitory protein), but CsA and the anti-CD147 antibody significantly inhibited these effects. Taken together, CsA in the presence of P.g-LPS might suppress MMP activities by blocking the CyPA/CD147 interaction that results in the inhibition of ERK1/2 and NF-κB signaling by interfering with the phosphorylation of ERK1/2 and IκB.

The Pros1/Tyro3 axis protects against periodontitis by modulating STAT/SOCS signalling.

Periodontitis, an oral inflammatory disease caused by periodontal pathogen infection, is the most prevalent chronic inflammatory disease and a major burden on healthcare. The TAM receptor tyrosine kinases (Tyro3, Axl and Mertk) and their ligands (Gas6 and Pros1) play a pivotal role in the resolution of inflammation and have been associated with chronic inflammatory and autoimmune diseases. In this study, we evaluated the effects of exogenous Pros1 in in vitro and in vivo models of periodontitis. We detected higher Pros1 but lower Tyro3 levels in inflamed gingival specimens of periodontitis patients compared with healthy controls. Moreover, Pros1 was mostly localized in the gingival epithelium of all specimens. In cultured human gingival epithelial cells (hGECs), Porphyromonas gingivalis LPS (p.g-LPS) stimulation down-regulated Pros1 and Tyro3. Exogenous Pros1 inhibited p.g-LPS-induced production of TNF-α, IL-6, IL-1ß, MMP9/2 and RANKL in a Tyro3-dependent manner as revealed by PCR, Western blot analysis, ELISA and gelatin zymography. Pros1 also restored Tyro3 expression down-regulated by p.g-LPS in hGECs. In rats treated with ligature and p.g-LPS, administration of Pros1 attenuated periodontitis-associated gingival inflammation and alveolar bone loss. Our mechanistic studies implicated SOCS1/3 and STAT1/3 as mediators of the in vitro and in vivo anti-inflammatory effects of Pros1. Collectively, the findings from this work supported Pros1 as a novel anti-inflammatory therapy for periodontitis.

The role of large-conductance, calcium-activated potassium channels in a rat model of trigeminal neuropathic pain.

BACKGROUND: Trigeminal neuralgia is a disorder of paroxysmal and severely disabling facial pain and continues to be a real therapeutic challenge. At present there are few effective drugs. Here the aim of this study was to investigate the role of BKCa channels in trigeminal neuropathic pain. METHODS: Rats were divided into two groups: a sham and a chronic constriction injury of infraorbital branch of trigeminal nerve (ION-CCI) group. Nociceptive behavior testing, immunohistochemistry, RT-PCR, Western blotting and whole-cell patch clamp recording were used. RESULTS: Relative to the sham group, rats with ION-CCI consistently displayed lower mechanical pain thresholds in the vibrissal pad region from day 6 to 42 after ION-CCI operation. ION-CCI induced a significant down-regulation of BKCa channels both in mRNA and protein levels in the ipsilateral trigeminal ganglion (TG), a lower threshold intensity of action potential, and decreased total BKCa currents in cultured TG neurons. TG target injection of NS1619 (20-100 µg), an opener of BKCa channels, dose-dependently increased the mechanical pain threshold, which was blocked by the BKCa channel inhibitor iberiotoxin (IbTX, 20 µg). NS1619 (10 µM) significantly increased the mean threshold intensities of action potentials in ION-CCI rats, while failing to affect those in the sham rats. The levels of phosphorylated extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinases (JNK) in TG were significantly increased after ION-CCI operation. The ERK1/2 antagonist U0126, p38 antagonist SB203580 and JNK antagonist SP600125 significantly reversed the facial mechanical allodynia in ION-CCI rats. However, the ERK1/2 antagonist U0126, p38 antagonist SB203580 but not JNK antagonist SP600125 significantly increased BKCa currents in ION-CCI TG neurons. CONCLUSIONS: Our results indicate the important involvement of mainly ERK and p38 MAPK pathways in modulating BKCa channels in ION-CCI TG neurons. BKCa channels represent a new therapeutic target for the clinical treatment of trigeminal neuropathic pain.

[BK(Ca) channel agonist NS1619 and Kv channel antagonist 4-AP on the facial mechanical pain threshold in a rat model of chronic constriction injury of the infraorbital nerve].

Trigeminal neuralgia is a paroxysmal disorder with severely disabling facial pain and thus continues to be a real therapeutic challenge. At present there are few effective drugs for treatment of this pain. The present study was aimed to explore the involvement of BK(Ca) channels and Kv channels in the mechanical allodynia in a rat model of trigeminal neuropathic pain. Here the effectiveness of drug target injection at the trigeminal ganglion through the infraorbital foramen was first evaluated by immunofluorescence and animal behavior test. Trigeminal neuropathic pain model was established by chronic constriction injury of the infraorbital nerve (ION-CCI) in rats. BK(Ca) channel agonist and Kv channel antagonist were administered into the trigeminal ganglion in ION-CCI rats and sham rats by the above target injection method, and the facial mechanical pain threshold was measured. The results showed that the drug could accurately reach the trigeminal ganglion by target injection which was more effective than that by the normal injection around infraorbital foramen. Rats suffered significant mechanical allodynia in the whisker pad of the operated side from 6 d to 42 d after ION-CCI. BK(Ca) channel agonist NS1619 significantly and dose-dependently attenuated the facial mechanical allodynia and increased the facial mechanical pain threshold in ION-CCI rats 15 d after operation. Kv antagonist 4-AP was able to reduce the threshold in ION-CCI rats when facial mechanical threshold was partly recovered and relatively stable on the 35th day after operation. These results suggest that BK(Ca) channel agonist NS1619 and Kv channel antagonist 4-AP can significantly affect the rats' facial mechanical pain threshold after ION-CCI. Activation of BK(Ca) channels may be related to the depression of the primary afferent neurons in trigeminal neuropathic pain pathways. Activation of Kv channels may exert a tonic inhibition on the trigeminal neuropathic pain.

Deletion of C9orf53 promotes the growth of head and neck squamous cell carcinoma and is associated with poor prognosis of patients with head and neck squamous cell carcinoma.

Head and neck squamous cell carcinoma (HNSCC) is the fifth most common carcinoma worldwide, and accounts for ~600,000 new cases every year. The information on the molecular carcinogenesis of HNSCC is very limited. In the present study, the role of C9orf53 in HNSCC was investigated. The levels of C9orf53 were assayed by reverse transcription-quantitative polymerase chain reaction. The levels of C9orf53 in cells were overexpressed by overexpression plasmid and inhibited by small-interfering RNA. Cell proliferation was assayed by MTT, and cell apoptosis was assessed by FACS analysis. It was demonstrated that C9orf53 deletion was associated with a decreased survival of patients. The level of C9orf53 in HNSCC tissues was lower compared with the matched normal tissues adjacent to tumors. A lower expression of C9orf53 promoted cell proliferation, and the overexpression of C9orf53 induced cell apoptosis. In conclusion, a low level of C9orf53 in HNSCC promoted the growth of HNSCC cells, which might be associated with the low survival rate of patients with HNSCC.

A novel videographic method for quantitatively tracking vibrissal motor recovery following facial nerve injuries in rats.

BACKGROUND: Previous studies of vibrissal movements employ either optoelectronic recording techniques in the head fixed rodent, or videographic recordings in freely moving animals. However, both approaches have shortcomings for quantitatively tracking the process of vibrissal motor recovery. NEW METHOD: A critical feature of our videographic method is to measure tagged vibrissae movements while leaving all others intact in body restrained rats without head fixation. Thirty two adult rats underwent facial nerve manipulation and testing. All animals underwent baseline preoperative whisking testing. In the experimental groups, the right facial nerve was either crushed, or transected and sutured. In the control groups, the left facial nerve underwent either sham surgery, or transection denervation. Whisking function was measured for the ensuing 2 to 12 weeks. Data were analyzed for whisking recovery. RESULTS: Baseline preoperative whisking testing showed that majority of free whisking on the both sides is synchronous and symmetric, which allows us to compare vibrissal motor data between intact and manipulated side after facial nerve injury. As expected, the recovery of whisking function following crush is better and earlier than that with transection and suture. COMPARISON WITH EXISTING METHOD(S): To our knowledge, this novel videographic method is a significant simplification over currently employed optoelectronic recording techniques and videographic methods. CONCLUSIONS: Our novel videographic method may be a powerful tool to investigate motor recovery from facial nerve manipulation in the rat model.

Serum cyclophilin A concentrations in renal transplant recipients receiving cyclosporine A: clinical implications for gingival overgrowth.

OBJECTIVE: The aim of this study was to investigate the clinical factors in relation to the cyclosporine A (CsA) induced gingival overgrowth (GO). STUDY DESIGN: Seventy-three participants were assigned as GO+ and GO-. Factors including demographic, pharmacological, gingival variables and the serum cyclophilin A (CyPA) concentration were analyzed. RESULTS: The occurrence of GO was 39.72%. Papillary bleeding index (PBI) had a significantly higher risk of GO than plaque index (PI), the ratio of CsA to CyPA, and serum CyPA concentration (odds ratio = 364.323, 25.791, 1.002, 0.096, respectively). The severity of GO correlated with PI, the ratio of CsA to CyPA, PBI, serum concentrations of CsA and CyPA (r = 0.366, 0.355, 0.344, 0.305, and -0.232, respectively). CONCLUSIONS: Since a cross-sectional study is not able to explain whether plaque and inflammation are the cause or consequence of GO, the ratio of CsA to CyPA may be a valuable marker for predicting GO.

Synchronous bilateral pleomorphic adenomas of the parotid gland.

Bilateral pleomorphic adenomas of the parotid gland are extremely rare, accounting for less than 0.2% of all parotid gland tumors. We present a rare case of a patient with synchronous bilateral pleomorphic adenomas of the parotid gland. A 27-year-old woman presented with a 5-month history of a slowly-growing, painless swelling of the right parotid gland. After a thorough bilateral parotid examination and magnetic resonance imaging evaluation, bilateral tumors of the parotid gland were found. The patient underwent surgical excision of the bilateral tumors in one surgical setting. Histopathological examination showed that both tumors were pleomorphic adenomas. The patient has been followed for 24 months without recurrence of tumors. Careful preoperative examination and radiological evaluation of bilateral parotid glands might be necessary for the early diagnosis of synchronous bilateral tumors. We suggest that treatment be individualized depending on the sizes and locations of tumors, and the surgical and neurological risks.

[Connection of trigeminal nerve and facial nerve branches and its clinical significance].

In recent years, many anatomical researches have showed that there are common and extensive connections between the trigeminal nerve and the facial nerve.They are briefly outlined as follows: (1) The infraorbital nerve communicates with buccal branch of the facial nerve. (2) The auriculotemporal nerve of the trigeminal nerve communicates with the buccal, zygomatic,temporal branches and the upper divisions of the facial nerve. (3) The supraorbital nerve communicates with the zygomatic and temporal branches of the facial nerve. (4) The mental nerve communicates with the marginal mandibular branch of the facial nerve. (5) The buccinator nerve communicates with the zygomatic, buccal and marginal mandibular branches. These communications between the trigeminal nerve and facial nerve are probably related to several clinical signs, for example,some trigeminal neuralgia patients are complicated by facial spasm, some patients appeared spontaneous partial functional recovery of mimetic muscles following surgical resection of a considerable segment of the facial nerve (including a portion of its main trunk and the peripheral plexus), etc. The purpose of this article was to review the anatomical features and clinical significance of the communications between the trigeminal nerve and the facial nerve.

[The neuromuscular toxicity and clinical application of adriamycin].

Adriamycin (doxorubicin) is commonly used in the treatment of malignant tumours. In recent years,retrograde adriamycin sensory ganglionectomy has been used in treatment of trigeminal neuralgia and has obtained good therapeutic effect. The mechanism of action, particularly, of the toxic effects with different medication methods and choice of doses for muscle cells and nerve cells is still unclear. This article reviewed the mechanism and feature of the toxicity of adriamycin effects on these cells and its advance in experimental study. The damage of adriamycin was highly selective and self-limited. The different effects of adriamycin with different administration routes and doses is also described. Adriamycin shows great potentiality in treatment of trigeminal neuralgia, facial spasm and some other neuromuscular diseases.

[Local drug injection for the treatment of trigeminal neuralgia].

Trigeminal neuralgia is the most common facial pain syndrome,which remains a difficult condition to manage due to lack of knowledge on pathophysiological mechanisms. Local drug injection is a minimally invasive and useful technique to manage trigeminal neuralgia, with a high success rate and low incidence of morbidity. Although a large number of drugs have been used to treat trigeminal neuralgia, there still remains a great challenge to reduce the recurrent rates and complications both intra- or post- injections.

[The expression and significance of osteopontin and its receptor CD44v6 in oral squamous carcinoma].

OBJECTIVE: To study the clinical significance of osteopontin (OPN) and its receptor CD44v6 expression in oral squamous cell carcinoma (OSCC). METHODS: OPN and CD44v6 expression were examined in OSCC (n=59) and normal oral mucosa (n=12) by EnVision method, the staining-grade were quantitatively studied by multiple functional image analyzer. Their expression grade of different clinical and pathological index were statistically studied. RESULTS: OPN expression grade was significantly higher in OSCC than that in normal oral mucosa (P<0.05). Significant deviation of OPN expression grade was found between different clinical stages, as well as between the groups with or without cervical lymph node metastasis. The group with cervical lymphnode metastasis had higher expression than that of the group without lymph node metastasis (P<0.05). However, there was no significant deviation between the expression grade in well-differentiated group and moderate or poorly differentiated group. The expression of CD44v6 showed no correlation with that of OPN, nor any difference between OSCC and normal oral mucosa. CONCLUSION: OPN over expression was found in OSCC, and the expression level has correlation with the clinical staging and with cervical lymph node metastasis status. CD44v6 expression showed no difference between OSCC and normal oral mucosa nor any correlation with that of OPN.

[Microvessel density and expression of vascular endothelial growth factor in adenoid cystic carcinoma of salivary gland].

OBJECTIVE: To discern whether the microvessel density (MVD) and the expression of vascular endothelial growth factor (VEGF) could be prognostic indicators for the metastasis and survival of patients with SACC after radical surgery. METHODS: After the follow-up of 31 primary patients with SACC treated with radical surgeries for at least 60 months, the paraffin-bedded sections of the those patients were stained immunohistochemically with anti-CD34-McAb and anti-VEGF-McAb, respectively. Subsequently, two pathologists double-blindly evaluated the sections stained with anti-CD34-McAb to determine the MVD values individually, as well as, measured the sections stained with the ani-VEGF-McAb to determine the OD values. Consequently the data were analyzed with the statistic software package detailed. RESULTS: The univariate statistical analysis showed that the pathological types of SACC, the TNM stages, MVD and OD values were all statistically significant variables for the survival of the patients with SACC (P = 0.047, 0.000, 0.000, 0.024). Furthermore, among those reliable variables, only MVD was more significant in the Cox proportional hazard model for the multivariate analyses (P = 0.000). The MVD values were statistically significantly higher, in the group with either death, or metastasis, or tumor-relapse, or worse pathological types, or advanced TNM stages, than their counterparts respectively(P = 0.029, 0.045, 0.019, 0.031, 0.00). On the other hand, the OD values were also statistically significantly higher, in the group with either death, or worse pathological types, or advanced TNM stages, than their counterparts respectively(P = 0.037, 0.013, 0.014). The results of Fisher exact test showed that the incidence of distant metastasis differed statistically significantly between the subgroups divided with the median of the MVD values (P = 0.032). The stepwise linear regression equation showed that the MVD value correlated positively with the OD value (P = 0.029). CONCLUSIONS: The MVD value and the expression level of VEGF all have statistically significant correlations with the survival of the patients with SACC, moreover, the MVD value is more significant as a prognostic indicator. The MVD value could also be a prognostic indicator for the incidence of the distant metastasis of patients with SACC. The over-expression of VEGF in SACC might contribute to the MVD values increasing.