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.

miR-146a participates in the regulation of cancer stemness of oral carcinoma cells.

Background/purpose: Increasing evidence regarded the existence of cancer stem cells (CSCs) as a leading cause of therapy failure and tumor relapse due to their self-renewal and differentiation abilities. Although ectopic overexpression of micro-RNAs (miRNAs) can modulate the cancer stemness and tumor development in oral cancer, their molecular mechanism is still unclear. Therefore, in the present study, we attempt to uncover the role of miR-146a in the maintenance of oral CSCs. Materials and methods: The expression of miR-146a was determined using qRT-PCR analysis. Aldehyde dehydrogenase (ALDH) enzymic activity and sphere formation assays were used to evaluate the cancer stemness and self-renewal, respectively. Functional assays, including migration/invasion Transwell and colony formation assay, were used to evaluate the aggressive abilities. Luciferase reporter assay was performed to validate the relationship between miR-146a and Numb. Results: In the present study, we reported an increased expression of miR-146a in the oral squamous cell carcinoma (OSCC) specimen, primary OSCC cells sphere, and high ALDH1 activity population within OSCC cells. Inhibition of miR-146a significantly suppressed the ALDH1 activity, self-renewal capacity, and aggressive abilities, including migration, invasion, and colony formation. Moreover, we demonstrated that Numb is a functional target of miR-146a in OSCC-CSCs. Notably, silencing of Numb could retrieve the self-renewal and migration impaired by knockdown of miR-146a. Conclusion: Our results indicate that miR-146a can regulate the cancer stemness in OSCC by modulating Numb, and hence miR-146a/Numb axis can serve as a potential target for oral cancer therapy.

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.

Improving the Diagnostic Performance by Adding Methylation Marker to Conventional Visual Examination in Identifying Oral Cancer.

BACKGROUND: Visual oral examination (VOE) is a conventional oral cancer screening method. This study aimed to evaluate the value of methylation marker to assist VOE in identifying oral epithelial dysplasia and oral squamous cell carcinoma (OED/OSCC) from non-cancerous lesions in a real-world situation. METHODS: 201 patients with high-risk personal habits who self-perceived oral anomaly were VOE examined, ZNF582 methylation (ZNF582m) tested, and histologically diagnosed. RESULTS: Among them, 132 patients (65.7%) were histologically diagnosed OED/OSCC. Using VOE, 56.1% OED/OSCC patients had possible oral cancer, whereas 37.7% non-OED/OSCC patients had leukoplakia. ZNF582m-positive was detected in 90.2% OED/OSCC patients and 44.9% non-OED/OSCC patients. Various logistic regression models were postulated to evaluate the diagnostic performance of conventional VOE and new strategies using ZNF582m. ROC analysis and its corresponding C-index demonstrated that either triage or co-testing models of VOE and ZNF582m could improve diagnostic performance and discriminative abilities compared with the VOE only approach. CONCLUSIONS: In conclusion, methylation marker test shows equivalent performance to an experienced judgment by oral maxillofacial surgeons and plays a significantly supplementary role in increasing the efficacy in identifying oral malignant lesions. ZNF582m may be an especially important tool for family physicians or general dentists to properly diagnose suspicious oral lesions.

miR-509 inhibits cancer stemness properties in oral carcinomas via directly targeting PlK1.

Background/purpose: Oral cancer is one of the common cancers worldwide. Emerging evidence has indicated that microRNAs (non-coding RNA molecules of approximately 22 nucleotides in length) are implicated in the regulation of cancer stemness. However, the functional role of microRNA-509 (miR-509) in the characteristics of oral cancer stem cells (CSCs) has not been unraveled. Materials and methods: The expression level of miR-509 in ALDH1+ and sphere oral CSCs was examined by qRT-PCR. The aldehyde dehydrogenase 1 (ALDH1) activity and CD44 expression were assessed using flow cytometry. Self-renewal, transwell migration, and colony formation assays were conducted to measure the CSC phenotypes. Besides, a luciferase reporter assay was used to confirm the direct interaction between miR-509 and its target polo-like kinase 1 (plk1). Results: We showed the expression of miR-509 was downregulated in the CSCs derived from oral cancer cells (SAS), and upregulation of miR-509 diminished the several CSCs features, including ALDH1 activity, self-renewal capacity, CD44 expression, migration, and colony-forming abilities. Moreover, the result from the luciferase reporter assay validated the direct binding of miR-509 to plk1. Conclusion: Our results suggest that the miR-509/plk1 axis may mediate the cancer stemness in oral cancer, and targeting this axis may attenuate the progression of oral cancer.

The functional roles of microRNAs in the pathogenesis of oral submucous fibrosis.

Oral potentially malignant disorders (OPMD) are lesions that may precede the onset of cancers in the oral cavity, and oral submucosal fibrosis (OSF) is one of the OPMD that is usually found in the buccal mucosa. Considerable effort has been made to elucidate the pathogenesis of OSF, and emerging evidence has suggested that microRNAs may play significant roles in the development of OSF. Several studies demonstrated that aberrant expression of miRNAs is also observed in the fibrotic BMFs (fBMFs) derived from OSF tissues. For instance, it has been shown that miR-10b, miR-21, and miR-1246 are significantly elevated, and miR-29b, miR-200b, and miR-200c are reduced in fBMFs. This review systematically summarizes the current knowledge regarding the aberrant expression of microRNAs, molecular mechanisms underlying oral fibrogenesis by the dysregulated microRNAs, and how the interaction between microRNAs and long non-coding RNAs contributes to the progression of OSF. An overview of the modes of action by these microRNAs will provide a fundamental basis for clinical application.

miR-21 promotes the fibrotic properties in oral mucosa through targeting PDCD4.

Background/purpose: Oral submucous fibrosis (OSF) has been regarded as a premalignant disorder of oral cancer, and myofibroblasts are the main cells that are responsible for pathological fibrosis. Hence, elucidation of the molecular mechanism underlying myofibroblast activation is important to treat OSF. MicroRNA-21 (miR-21) is a well-known fibrosis non-coding RNA, and its role in the development of OSF remains largely unclear. Materials and methods: Luciferase reporter assay was used to confirm the direct interaction between miR-21 and its target programmed cell death 4 (PDCD4). The expression level of PDCD4 in OSF was examined by qRT-PCR. Myofibroblast activities were assessed by collagen gel contraction and transwell migration assays. Results: Our result validated the direct binding of miR-21 to PDCD4. We showed the expression of PDCD4 was downregulated in OSF specimens and negatively correlated with miR-21. Our results suggested that overexpression of PDCD4 in fibrotic buccal mucosal fibroblasts (fBMFs) mitigated the myofibroblast activities, including collagen gel contractility and migration capacity. Moreover, we showed miR-21 contributed to myofibroblast activation of BMFs through repression of PDCD4. Conclusion: Our results suggest that the miR-21/PDCD4 axis mediates the myofibroblast activation of BMFs, and targeting this axis may exert an anti-fibrosis effect.

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.

LINC00084/miR-204/ZEB1 Axis Mediates Myofibroblastic Differentiation Activity in Fibrotic Buccal Mucosa Fibroblasts: Therapeutic Target for Oral Submucous Fibrosis.

Oral submucosal fibrosis (OSF) is a precancerous condition in the oral cavity and areca nut consumption has been regarded as one of the etiologic factors implicated in the development of OSF via persistent activation of buccal mucosal fibroblasts (BMFs). It has been previously reported that an epithelial to mesenchymal transition (EMT) factor, ZEB1, mediated the areca nut-associated myofibroblast transdifferentiation. In the current study, we aimed to elucidate how areca nut affected non-coding RNAs and the subsequent myofibroblast activation via ZEB1. We found that long non-coding RNA LINC00084 was elicited in the BMFs treated with arecoline, a major alkaloid of areca nut, and silencing LINC00084 prevented the arecoline-induced activities (such as collagen gel contraction, migration, and wound healing capacities). The upregulation of LINC00084 was also observed in the OSF tissues and fibrotic BMFs (fBMFs), and positively correlated with several fibrosis factors. Moreover, we showed knockdown of LINC00084 markedly suppressed the myofibroblast features in fBMFs, including myofibroblast phenotypes and marker expression. The results from the luciferase reporter assay confirmed that LINC00084 acted as a sponge of miR-204 and miR-204 inhibited ZEB1 by directly interacting with it. Altogether, these findings suggested that the constant irritation of arecoline may result in upregulation of LINC00084 in BMFs, which increased the ZEB1 expression by sequestering miR-204 to induce myofibroblast transdifferentiation.

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.

Addition of oyster shell to enhance organic matter degradation and nitrogen conservation during anaerobic digestate composting.

Anaerobic digested residue (DR) is the main by-product from biogas plants, and it is predominantly used as organic fertilizer after composting. To resolve the problems of long duration and nitrogen loss in conventional composting, bulking agents are always added during the composting process. In this study, oyster shell (OS) was used as a bulking agent for DR composting. Four treatments were conducted by mixing DR and OS at different concentrations (0%, 10%, 20% and 30%, based on wet weight) and then composting the mixtures for 40 days. The results showed that the organic matter (OM) degradation efficiency was enhanced by 5.62%, 12.15% and 16.98% with increasing amounts of OS addition. The increased content of microbial biomass carbon in the compost indicated a suitable living environment for aerobic microbes with added OS, which could explain the increased OM degradation efficiency. Compared with the control, the NH3 emissions in the treatments with 10%, 20% and 30% OS were decreased by 13.81%, 33.33% and 53.76%, respectively. The increase in total nitrogen content in the compost is probably due to the absorption of NH3 by OS. Results indicated that OS is a suitable bulking agent for DR composting and that the addition of 20-30% OS can significantly enhance composting performance.

Positive Feedback Loop of SNAIL-IL-6 Mediates Myofibroblastic Differentiation Activity in Precancerous Oral Submucous Fibrosis.

Oral submucosal fibrosis (OSF) is a premalignant disorder of the oral cavity, and areca nut chewing is known to be a major etiological factor that could induce epithelial to mesenchymal transition (EMT) and activate buccal mucosal fibroblasts (BMFs). However, this detailed mechanism is not fully understood. In this study, we showed that the upregulation of Snail in OSF samples and fibrotic BMFs (fBMFs) may result from constant irritation by arecoline, a major alkaloid of the areca nut. The elevation of Snail triggered myofibroblast transdifferentiation and was crucial to the persistent activation of fBMFs. Meanwhile, Snail increased the expression of numerous fibrosis factors (e.g., α-SMA and collagen I) as well as IL-6. Results from bioinformatics software and a luciferase-based reporter assay revealed that IL-6 was a direct target of Snail. Moreover, IL-6 in BMFs was found to further increase the expression of Snail and mediate Snail-induced myofibroblast activation. These findings suggested that there was a positive loop between Snail and IL-6 to regulate the areca nut-associated myofibroblast transdifferentiation, which implied that the blockage of Snail may serve as a favorable therapeutic strategy for OSF treatment.

Gold-Catalyzed Oxidations of 1,3-Diynamides with C(1) versus C(3) Regioselectivity: Catalyst-Dependent Oxidative Cyclizations in the C(3) Oxidation.

This work reports two distinct paths in catalytic oxidations of 1,3-diynamides with 8-methylquinoline oxide. A typical C(1) regioselectivity was observed for aryl-substituted 1,3-diyn-1-amides, whereas an unexpected C(3) regioselectivty occurred for 5-hydroxy1,3-diyn-1-amides. We focused on the C(3) oxidations of 5-hydroxy1,3-diyn-1-amides because we observed two oxidative cyclizations of the same substrates to yield 2-aminomethylenefuran-3(2H)-ones and 2-amino-4H-pyran-4-ones using AuCl3 and a cationic gold catalyst, respectively. Density functional theory calculations were performed to rationalize the C(3) regioselectivity on 5-hydroxy1,3-diyn-1-amides.

Improved nitrogen conservation capacity during composting of dairy manure amended with oil shale semi-coke as the porous bulking agent.

Oil shale semi-coke is the solid waste produced from the retorting process of oil shale, which may cause pollution to the environment without reasonable disposing. In this study, semi-coke was used as the bulking agent during composting to accelerate biodegradation of the organics as well as decrease the nitrogen loss. Results showed that the addition of semi-coke could accelerate biodegradation of the organics, with a raise in the organic matter loss from 44.99 % to 47.05 % compared with the control. Furthermore, the nitrogen loss significantly decreased from 40.00%-14.70 % in the treatment added with semi-coke due to less emission of NH3 and much more transformation of NH4+-N to NO3--N by nitrification, which could be explained by the increasing abundance of ammonia-oxidizing bacteria and archaea at the late composting stage and drastic shift of the microbial community like Chloroflexi, Firmicutes and Actinobacteria. After the composting cycle, the maturity of the produced compost was elevated greatly in the treatments amended with semi-coke. The result of PAHs detection suggested that there were low PAHs content in the raw oil shale semi-coke and they could be removed effectively to within the range for land application by composting especially when the surfactant was added.

Succession of the bacterial community and functional characteristics during continuous thermophilic composting of dairy manure amended with recycled ceramsite.

This study aimed to investigate variations of bacterial community and functional characteristics during the continuous thermophilic composting (CTC). Also their differences were discussed when amended with ceramsite and recycled ceramsite as the porous bulking agent. Results showed that the bacterial community shifted greatly and bacterial diversity increased as the CTC proceeded. Firmicutes and Chloroflexi was one of the major phyla at the active and late phase respectively. While Actinobacteria was the dominant phyla during the whole CTC. With the addition of ceramsite and recycled ceramsite, no significant difference was found of the overall bacterial variation trends. But the major phyla of Chloroflexi and Actinobacteria and major genes related to amino acid metabolism and carbohydrate metabolism increased significantly, especially when the recycled ceramsite was added. Redundancy analysis indicated that the succession of bacterial community was tightly related with the pH, water soluble organic carbon, NH4+-N, organic matter and germination index.