Is OLP potentially malignant? A clue from ZNF582 methylation.

OBJECTIVE: Whether oral lichen planus (OLP) was potentially malignant remains controversial. Here, we examined associations of ZNF582 methylation (ZNF582m ) with OLP lesions, dysplastic features and squamous cell carcinoma (OSCC). MATERIALS AND METHODS: This is a case-control study. ZNF582m was evaluated in both lesion and adjacent normal sites of 42 dysplasia, 90 OSCC and 43 OLP patients, whereas ZNF582m was evaluated only in one mucosal site of 45 normal controls. High-risk habits affecting ZNF582m such as betel nut chewing and cigarette smoking were also compared in those groups. RESULTS: OLP lesions showed significantly lower ZNF582m than those of dysplasia and OSCC. At adjacent normal mucosa, ZNF582m increased from patients of OLP, dysplasia, to OSCC. In addition, ZNF582m at adjacent normal sites in OLP patients was comparable to normal mucosa in control group. Dysplasia/OSCC patients with high-risk habits exhibited significantly higher ZNF582m than those without high-risk habits. However, ZNF582m in OLP patients was not affected by those high-risk habits. CONCLUSIONS: OLP is unlikely to be potentially malignant based on ZNF582m levels. ZNF582m may also be a potential biomarker for distinguishing OLP from true dysplastic features and OSCC, and for monitoring the malignant transformation of OLP, potentially malignant disorders with dysplastic features and OSCC.

Use of magnetic powder to effectively improve the denitrification employing the activated sludge fermentation liquid as carbon source.

Nitrogen removal is often limited in municipal wastewater treatment due to the lack of sufficient carbon source. Utilizing volatile fatty acids (VFAs) from waste activated sludge (WAS) fermentation broth as a carbon source is an ideal alternative to reduce the cost for wastewater treatment plants (WWTPs) and improve denitrification efficiency simultaneously. In this study, an anaerobic system was applied for simultaneous denitrification and WAS fermentation and the addition of magnetic microparticles (MMP) were confirmed to enhance both denitrification and WAS fermentation. Firstly, the addition of MMP increased the nitrate reduction rate by over 25.36% and improve the production of N2. Additionally, the equivalent chemical oxygen demand (COD) of the detected VFAs increased by 7.06%-14.53%, suggesting that MMP promoted the WAS fermentation. The electron transfer efficiency of denitrifies was accelerated by MMP via electron-transporting system (ETS) activity and cyclic voltammetry (CV) experiments, which might result in the promotional denitrification and WAS fermentation performance. Furthermore, the high-throughput sequencing displayed that, MMP enriched key microbes capable of degrading the complex organics (Chloroflexi, Synergistota and Spirochaetota) as well as the typical denitrifies (Bacteroidetes_vadinHA17 and Denitratisoma). Therefore, this study provides a novel strategy to realize simultaneous WAS utilization and denitrification for WWTPs.

Improved fertilization efficiency of manure compost obtained by using oil shale semicoke as the bulking agent.

Oil shale semicoke is a kind of solid waste produced during the retorting process of oil shale, which could cause environmental pollution without reasonable disposing. In our previous study, the abandoned semicoke was recycled as bulking agent to reduce the nitrogen loss and greenhouse gases emission during composting. But influences of the obtained semicoke-blended compost on soil properties and plant growth remained unclear, which would be discussed in this study. Through leaching experiments, it was found that the N/P/K retention capacity of soil mixed with semicoke-blended compost significantly increased for the good nutrients sorption capacity of oil shale semicoke. Subsequently, germination test showed the germination index of semicoke-blended compost could attain 120%, implying its low phytotoxicity. And pot experiments exhibited the biomass of cress and Brassica rapa significantly increased by 2-4 times when applying semicoke-blended compost as fertilizer, exhibiting its great benefits to plants. For the increase of crop yield, it was closely related to their elevated nutrients uptake efficiency, also might be related to the improved soil microbial community and activity as the microbial analysis indicated. Finally, results of pollutant detection showed the concentration of polycyclic aromatic hydrocarbons, Cr, As, Cd and Pb in the mature semicoke-blended compost obtained through composting was 2.82, 95.30, 5.95, 0.34 and 14.45 mg kg-1 respectively, meeting the standard for soil application. The research suggests composting could be an effective method for the harmless disposing and resource recycling of oil shale semicoke waste.

Effective dose and radiation risk under 640-slice abdominal computed tomography examination without contrast medium injection.

BACKGROUND: Medical imaging plays a crucial role in modern medicine. In order to provide fast and accurate medical diagnosis, computed tomography (CT) is a commonly used tool in radiological examinations, and 640-slice CT is the most advanced CT imaging modality. OBJECTIVE: To evaluate the radiation dose and the risk under 640-slice abdominal CT examination. METHODS: Examinations were performed using a 640-slice CT scanner on an Alderson-Rando anthropomorphic phantom. The used scanning acquisition parameters were the same as those used on abdominal examination without contrast medium injection in clinical practice. To measure the absorbed doses, optically stimulated luminescence dosimeters (OSLDs) were put into liver, stomach, bladder, gonads, colon, small intestine, bone marrow, and skin. RESULTS: According to the 1990 Recommendations of the International Commission on Radiological Protection (ICRP Publication 60), the calculated effective doses received from this examination were 0.90 mSv in males and 0.89 mSv in females. According to the 2007 Recommendations of the International Commission on Radiological Protection (ICRP Publication 103), the calculated effective dose received from this examination was 0.83 mSv in both sexes. CONCLUSIONS: Radiation doses obtained from the abdominal 640-slice CT examination are lower than the yearly cumulative doses received from natural radiation, revealing there is no deterministic effect and radiation risk is relatively low; therefore, this CT examination is considered safe.

Oral microbial dysbiosis and its performance in predicting oral cancer.

Dysbiosis of oral microbiome may dictate the progression of oral squamous cell carcinoma (OSCC). Yet, the composition of oral microbiome fluctuates by saliva and distinct sites of oral cavity and is affected by risky behaviors (smoking, drinking and betel quid chewing) and individuals' oral health condition. To characterize the disturbances in the oral microbial population mainly due to oral tumorigenicity, we profiled the bacteria within the surface of OSCC lesion and its contralateral normal tissue from discovery (n = 74) and validation (n = 42) cohorts of male patients with cancers of the buccal mucosa. Significant alterations in the bacterial diversity and relative abundance of specific oral microbiota (most profoundly, an enrichment for genus Fusobacterium and the loss of genus Streptococcus in the tumor sites) were identified. Functional prediction of oral microbiome shown that microbial genes related to the metabolism of terpenoids and polyketides were differentially enriched between the control and tumor groups, indicating a functional role of oral microbiome in formulating a tumor microenvironment via attenuated biosynthesis of secondary metabolites with anti-cancer effects. Furthermore, the vast majority of microbial signatures detected in the discovery cohort was generalized well to the independent validation cohort, and the clinical validity of these OSCC-associated microbes was observed and successfully replicated. Overall, our analyses reveal signatures (a profusion of Fusobacterium nucleatum CTI-2 and a decrease in Streptococcus pneumoniae) and functions (decreased production of tumor-suppressive metabolites) of oral microbiota related to oral cancer.

Gases emission during the continuous thermophilic composting of dairy manure amended with activated oil shale semicoke.

NH3 and greenhouse gases emission are big problems during composting, which can cause great nitrogen nutrient loss and environmental pollution. This study investigated effects of the porous bulking agent of oil shale semicoke and its activated material on the gases emission during the continuous thermophilic composting. Results showed addition of semicoke could significantly reduce the NH3 emission by 74.65% due to its great adsorption capacity to NH4+-N and NH3, further the effect could be enhanced to 85.92% when utilizing the activated semicoke with larger pore volume and specific surface area. In addition, the CH4 emission in the semicoke and activated semicoke group was also greatly mitigated, with a reduction of 67.23% and 87.62% respectively, while the N2O emission was significantly increased by 93.14% and 100.82%. Quantification analysis of the functional genes found the abundance of mcrA was high at the massive CH4-producing stage and the archaeal amoA was dominant at the N2O-producing stage in all the composting groups. Correlation and redundancy analysis suggested there was a positive correlation between the CH4 emission and mcrA. Addition of semicoke especially activated semicoke could reduce the CH4 production by inhibiting the methanogens. For the NH3 and N2O, it was closely related with the nitrification process conducted by archaeal amoA. Addition of semicoke especially activated semicoke was beneficial for the growth of ammonia-oxidizing archaea, causing the less NH4+-N transformation to NH3 but more N2O emission.

LncRNA LINC00974 activates TGF-ß/Smad signaling to promote oral fibrogenesis.

BACKGROUND: Oral submucous fibrosis (OSF) is a progressive scarring disease and has been considered as a premalignant condition of the oral cavity. However, the detailed molecular mechanisms underlying the pathogenesis of OSF are still unclear. METHOD: Here, we examined the expression of a novel long non-coding RNA LINC00974 in OSF and investigated its function role in myofibroblast transdifferentiation. Phenotypic analyses, including collagen gel contraction, migration, invasion and wound healing assays, were used to assess the myofibroblast activities following overexpression or inhibition of LINC00974. RESULTS: We found that the expression of LINC00974 in OSF tissues or myofibroblasts was aberrantly upregulated, and there was a positive correlation between LINC00974 and myofibroblast markers. Our results showed that inhibition of LINC00974 suppressed the myofibroblast activities, while overexpression of LINC00974 increased the activation. We demonstrated that the expression levels of α-SMA, α-1 type I collagen, fibronectin were downregulated in the LINC00974-inhibited myofibroblasts. Additionally, the TGF-ß secretion and phosphorylated Smad2 expression were also repressed in the LINC00974-inhibited myofibroblasts. We further demonstrated that silence of LINC00974 prevented the arecoline-induced myofibroblast activation, and LINC00974-increased myofibroblast activities were via TGF-ß pathway. CONCLUSION: Altogether, these findings suggested that arecoline-increased myofibroblast transdifferentiation was via LINC00974-mediated activation of TGF-ß signaling.

Arctigenin Reduces Myofibroblast Activities in Oral Submucous Fibrosis by LINC00974 Inhibition.

Oral submucous fibrosis (OSF) is an oral precancerous condition associated with the habit of areca nut chewing and the TGF-ß pathway. Currently, there is no curative treatment to completely heal OSF, and it is imperative to alleviate patients' symptoms and prevent it from undergoing malignant transformation. Arctigenin, a lignan extracted from Arctium lappa, has been reported to have a variety of pharmacological activities, including anti-fibrosis. In the present study, we examined the effect of arctigenin on the cell proliferation of buccal mucosal fibroblasts (BMFs) and fibrotic BMFs (fBMFs), followed by assessment of myofibroblast activities. We found that arctigenin was able to abolish the arecoline-induced collagen gel contractility, migration, invasion, and wound healing capacities of BMFs and downregulate the myofibroblast characteristics of fBMFs in a dose-dependent manner. Most importantly, the production of TGF-ß in fBMFs was reduced after exposure to arctigenin, along with the suppression of p-Smad2, α-smooth muscle actin, and type I collagen A1. In addition, arctigenin was shown to diminish the expression of LINC00974, which has been proven to activate TGF-ß/Smad signaling for oral fibrogenesis. Taken together, we demonstrated that arctigenin may act as a suitable adjunct therapy for OSF.

Chemosensitizing effect of honokiol in oral carcinoma stem cells via regulation of IL-6/Stat3 signaling.

Oral squamous cell carcinoma (OSCC) is one of the most common cancers worldwide with poor prognosis. Numerous studies have attempted to explore alternative regimens aimed at reducing cancer stem cells (CSCs) without compromising the efficacy of conventional chemoradiotherapy. The present study sought to assess the effect of a natural compound honokiol on the reduction of elevated cancer stemness, metastatic capacity, and chemoresistance of oral carcinoma stem cells (OCSCs). Our results demonstrated that honokiol attenuated the cell survival and self-renewal of OCSCs in a dose-dependent manner. Moreover, honokiol downregulated the expression of 2 selective markers of OCSCs, ALDH1, and CD44, as well as the migration and invasion abilities, indicating its potential to suppress cancer stemness. We showed that honokiol reduced the secretion of IL-6 and phosphorylation of STAT3, and the honokiol-inhibited self-renewal, invasion and colony formation were reversed by administration of IL-6. Most importantly, our data demonstrated that honokiol was able to potentiate the effect of Cisplatin, leading to a lower proportion of OCSCs and the decreased cancer stemness features. Taken together, this study demonstrated the benefits of utilizing honokiol as an adjunct therapy for OSCC treatment.

Hinokitiol ablates myofibroblast activation in precancerous oral submucous fibrosis by targeting Snail.

Oral submucous fibrosis (OSF) is a precancerous condition with symptoms of limited mouth opening and areca nut chewing habit has been implicated in its pathogenesis. Hinokitiol, a natural tropolone derived from Chamacyparis taiwanensis, has been reported to improve oral lichen planus and inhibit various cancer cells. Here, we showed that hinokitiol reduced the myofibroblast activities in fBMFs and prevented the arecoline-induced transdifferentiation. Treatment of hinokitiol dose-dependently downregulated the myofibroblast markers as well as various EMT transcriptional factors. In particular, we identified that Snail was able to bind to the E-box in the α-SMA promoter. Our data suggested that exposure of fBMFs to hinokitiol mitigated the hallmarks of myofibroblasts, while overexpression of Snail eliminated the effect of hinokitiol. These findings revealed that the inhibitory effect of hinokitiol on myofibroblasts was mediated by repression of α-SMA via regulation of Snail and showed the anti-fibrotic potential of hinokitiol in the treatment of OSF.

miR-200c inhibits the arecoline-associated myofibroblastic transdifferentiation in buccal mucosal fibroblasts.

BACKGROUND/PURPOSE: MicroRNA-200c (miR-200c) recently emerged as an important regulator of tumorigenesis and cancer metastasis, however, its role in regulating oral submucous fibrosis (OSF) remains unknown. In this study, we investigated the functional role of miR-200c in myofibroblastic differentiation activity and identified its potential target. METHODS: qRT-PCR was applied to assess the expression of miR-200c in OSF tissues and fibrotic buccal mucosal fibroblasts (fBMFs). Arecoline, a major areca nut alkaloid, was utilized to explore whether the expression of miR-200c would alter following stimulation. Collagen gel contraction, migration and invasion capabilities were examined in arecoline-stimulated BMFs as wells as in fBMFs. Luciferase reporter assay was conducted to show the relationship between miR-200c and ZEB1. RESULTS: Our results showed that the expression of miR-200c was downregulated in OSF specimen and fBMFs. Arecoline treatment dose-dependently reduced the relative expression of miR-200c in normal BMFs. Overexpression of miR-200c impeded the arecoline-induced collagen gel contraction, migration, invasion and wound healing capacities. Moreover, ectopic expression of miR-200c in fBMFs successfully reduced the increased collagen gel contractility and invasion abilities. Our results demonstrated that ZEB1 was a direct target of miR-200c, and overexpression of miR-200c inhibited the expression of ZEB1 and α-SMA. CONCLUSION: These findings suggest that downregulation of miR-200c in OSF may be involved in the pathogenesis of areca nut-associated OSF through regulation of ZEB1.

miR-145 mediates the anti-cancer stemness effect of photodynamic therapy with 5-aminolevulinic acid (ALA) in oral cancer cells.

5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) has been used in the treatment of various precancerous and malignant lesions. Our previous work has demonstrated that ALA-PDT possesses the potential to serve as an adjuvant therapy against head and neck cancer via eliminating the cancer stem cells (CSCs) property. This study aimed to further investigate the possible molecular mechanism underlying the effect of ALA-PDT. Our results revealed that ALA-PDT upregulated the expression of microRNA-145 (miR-145) in two oral cancer cell lines. Overexpression of miR-145 in oral CSCs further enhanced the treatment effect of ALA-PDT with lower self-renewal, invasion capacities and reduced CD44 expression, while inhibition of miR-145 exhibited the opposite phenomena. These findings suggest that the anti-CSCs effect of ALA-PDT is due to an elevation of miR-145.

LncRNA GAS5-AS1 inhibits myofibroblasts activities in oral submucous fibrosis.

BACKGROUND/PURPOSE: Emerging research findings suggest that long non-coding RNAs (lncRNAs) are key regulators to fibrosis formation. Nevertheless, the role of lncRNA GAS5-AS1 in the progression of precancerous oral submucous fibrosis (OSF) remains to be elucidated. METHODS: Quantitative real-time PCR were used to examine the expression of GAS5-AS1 in OSF tissues. The activities of myofibroblasts, including collagen contractility and cell migration, as well as the marker α-smooth muscle actin (SMA) were assessed following overexpression of GAS5-AS1. Also, we analyzed the expression of Smad activity in order to gain insight into the downstream regulator. RESULTS: The level of GAS5-AS1 was found significantly downregulated in the OSF tissues and fibrotic buccal mucosal fibroblasts (fBMFs). Ectopic expression of GAS5-AS1 significantly reduced the abilities of collagen gel contraction and migration in fBMFs or arecoline-treated BMFs. Moreover, we have shown that overexpression of GAS5-AS1 inhibited the expression of p-Smad and the marker of myofibroblasts. CONCLUSION: We showed the reduced expression of GAS5-AS1 in OSF tissues and demonstrated its effect on the myofibroblast activities and the level of p-Smad and α-SMA, indicating its potential contribution in OSF pathogenesis.

Downregulation of miR-1 enhances tumorigenicity and invasiveness in oral squamous cell carcinomas.

BACKGROUND/PURPOSE: Cumulative evidence suggest that microRNAs (miRNAs) function as biosignatures of oral squamous cell carcinomas (OSCC). However, the functional roles of miR-1 as well as its downstream targets in the regulation of tumorigenicity in OSCC remain unclear. METHODS: miRNAs RT-PCR analysis was performed to identify miR-1 as a putative candidate on mediating invasiveness of OSCC cells. Consequently, we elucidated the tumorigenicity of OSCC cells with miR-1 downregulation or overexpression, respectively. Finally, miR-1 on OSCC tumor tissues was examined. RESULTS: miR-1 levels were significantly downregulated in the malignant OSCC cells. Overexpression of miR-1 significantly reduced migration/invasiveness of OSCC cells. In addition, overexpression of miR-1 decreased cancer stem cells properties. Conversely, downregulation of miR-1 promotes migration and invasiveness in OSCC cells. We have shown that miR-1 is able to target Slug, suppressing their expression. Clinically, lower miR-1 expression was found in patients with advanced nodal metastasis OSCC. CONCLUSION: miR-1 as novel biosignatures in OSCC lymph node metastatic patients, supporting the development of novel strategies for OSCC treatment.

Sulforaphane targets cancer stemness and tumor initiating properties in oral squamous cell carcinomas via miR-200c induction.

BACKGROUND/PURPOSE: Cancer stem cells (CSCs) are deemed as the driving force of tumorigenesis in oral squamous cell carcinomas (OSCCs). In this study, we investigated the chemotherapeutic effect of sulforaphane, a dietary component from broccoli sprouts, on targeting OSCC-CSCs. METHODS: The effect of sulforaphane on normal oral epithelial cells (SG) and sphere-forming OSCC-CSCs isolated from SAS and GNM cells was examined. ALDH1 activity and CD44 positivity of OSCC-CSCs with sulforaphane treatment was assessed by flow cytometry analysis. In vitro and in vivo tumorigenicity assays of OSCC-CSCs with sulforaphane treatment were presented. RESULTS: We observed that the sulforaphane dose-dependently eliminated the proliferation rate of OSCC-CSCs, whereas the inhibition on SG cells proliferation was limited. Cancer stemness properties including self-renewal, CD44 positivity, and ALDH1 activity were also decreased in OSCC-CSCs with different doses of sulforaphane treatment. Moreover, sulforaphane treatment of OSCC-CSCs decreased the migration, invasion, clonogenicity, and in vivo tumorigenicity of xenograghts. Sulforaphane treatment resulted in a dose-dependent increase in the levels of tumor suppressive miR200c. CONCLUSION: These lines of evidence suggest that sulforaphane can suppress the cancer stemness and tumor-initiating properties in OSCC-CSCs both in vitro and in vivo.

Intraosseous verrucous carcinoma arising from an infected dentigerous cyst-A case report.

Intraosseous verrucous carcinoma (IOVC) arising from an odontogenic cyst is extremely rare. We report a case of intraosseous verrucous carcinoma in a 74-year-old male who presented with a left mandibular swelling with recurrent pus discharge from gingiva of tooth #35. Panoramic radiography revealed an impacted tooth #34 and a large well-defined, radiolucent lesion surrounding the crown of tooth #34. The clinical diagnosis was an infected dentigerous cyst. Surgical excision of the cyst together with extraction of tooth #34 was performed. Histopathological examination showed proliferation of hyperparakeratotic stratified squamous cyst lining epithelium and down-growth of broad and bulbous epithelial ridges with pushing border invasion into the fibrous cystic wall. A verrucous carcinoma arising from an infected dentigerous cyst was diagnosed. There was no recurrence of the tumor 5 months after surgery.

Novel flocking materials as biocarriers in moving bed biofilm reactor for improving simultaneous nitrification and denitrification performance.

Biocarrier is the key factor for the stable operation of moving bed biofilm reactor (MBBR). To achieve efficient simultaneous nitrification and denitrification (SND), this study provided novel flocking materials as biocarriers. The biofilm formation experiment showed that longer flocking carrier was more conducive to biomass accumulation, resulting in greater oxygen uptake rate. The continuous operation results showed that the total nitrogen removal and SND performance of the MBBR with the addition of 5.0 mm flocking carriers reached 52.0 % and 70.5 %, respectively, which were 29.1 % and 33.3 % greater than those of the control. Compared with those in suspended sludge, the extracellular polymeric substances and protein components in the biocarrier were more abundant. Furthermore, the relative abundance of genera related to denitrification and the nitrogen metabolic sequence improved with the addition of the novel flocking biocarriers. This study demonstrated the effectiveness of novel flocking fillers in improving the performance of MBBR.

Simultaneous denitrification enhancement and sludge reduction based on novel suspended carrier modified using activated carbon and magnetite at low carbon/nitrogen ratio.

A novel suspended carrier was prepared by sticking activated carbon (AC) and magnetite (Fe3O4) onto polypropylene slices. Although this carrier could not reverse the decreased denitrification capacity trends under anoxic conditions at an influent carbon/nitrogen (C/N) ratio of 2, it enhanced denitrification by stimulating sludge reduction and accelerating electron transfer to certain extent. The carrier stuck by mixed AC/Fe3O4 exhibited better performance in terms of sludge reduction, extracellular polymeric substances (EPS) secretion, and denitrification than that merely stuck by AC and Fe3O4 at an influent C/N ratio of 2. The carrier stuck by mixed AC/Fe3O4 increased the total nitrogen removal efficiency by 24.6 % ± 12.5 % in a 72-h denitrification batch experiment compared to the common polypropylene carrier. Moreover, the carrier improved EPS secretion and nitrogen metabolism and promoted the growth of Trichococcus and some denitrifying genera. This study provides a reference for the treatment of low C/N ratio sewage.

Use of reconstruction plates prebent on three-dimensional models to reduce the complications of mandibular reconstruction.

Background/purpose: Though the gold standard method for mandible reconstruction of the defect from segmental mandibulectomy is by osseous flap or graft, using reconstruction plates is still indicated in some cases. Traditionally, the plate is bended immediately after the segmental mandibulectomy by freehand. However, it's difficult to fit well to the original position of mandible, which may result in more complications. This study therefore aimed to investigate whether using prebent plates on computer-aided 3D printing models could reduce the complication rate. Materials and methods: Patients who received mandible reconstruction by reconstruction plate from 2018 to 2022 were enrolled and evaluated in this study. The data, including demographics, indications for surgery, pre-existed preoperative and postoperative therapies, classification of defects, and postoperative outcomes were collected and analyzed. Results: A total of 52 patients were enrolled in our study. The prebent group exhibited a significantly lower complication rate than that of the immediately bent group (P = 0.012). Other risk factors of plate complications included postoperative adjuvant radiotherapy (P = 0.017) and previous surgery (P = 0.047). The complication-free survival rate was also better in the prebent group in a 3-year follow-up period (P = 0.012). Conclusion: Prebent plates on computer-aided printing models proved to be an effective approach to reduce the complications for mandibular reconstruction in segmental mandibulectomy.