Advances in the Prophylaxis of Respiratory Infections by the Nasal and the Oromucosal Route: Relevance to the Fight with the SARS-CoV-2 Pandemic.

In this time of COVID-19 pandemic, the strategies for prevention of the infection are a primary concern. Looking more globally on the subject and acknowledging the high degree of misuse of protective face masks from the population, we focused this review on alternative pharmaceutical developments eligible for self-defense against respiratory infections. In particular, the attention herein is directed to the nasal and oromucosal formulations intended to boost the local immunity, neutralize or mechanically "trap" the pathogens at the site of entry (nose or mouth). The current work presents a critical review of the contemporary methods of immune- and chemoprophylaxis and their suitability and applicability in topical mucosal dosage forms for SARS-CoV-2 prophylaxis.

A randomised clinical trial on the effect of oral antimicrobial sprays in institutionalised elders.

OBJECTIVE: To compare the effectiveness of chlorine dioxide, chlorhexidine and placebo sprays in improving oral hygiene among institutionalised elders. BACKGROUND: Available evidence suggests that oral sprays may be an effective alternative delivery method for plaque control; however, few studies have evaluated antimicrobial agents other than chlorhexidine. MATERIALS AND METHODS: A total of 228 elders across 11 nursing homes in Hong Kong were recruited into the clinical trial. Participants were randomly allocated into one of the following groups: 0.1% pH-balanced chlorine dioxide spray, 0.2% chlorhexidine spray or sterile water spray (placebo control), once daily. Dental plaque, gingival bleeding and other clinical oral health outcomes were assessed at baseline, 3 and 6 months. Participant acceptability of the interventions was assessed at the end of the clinical trial. RESULTS: Review assessments were conducted for 135 elders at 6 months. Significantly greater reductions in plaque index scores were observed with the chlorhexidine spray (0.4) and chlorine dioxide spray (0.3) than the placebo spray (0.1). While significant reductions in gingival bleeding scores were observed within the chlorhexidine (7.4), chlorine dioxide (7.5) and placebo (5.3) sprays after 6 months, change scores were not significantly different between groups. Significantly greater increases in the levels of staining were observed in the chlorhexidine spray group (-0.1) than the chlorine dioxide (0.0) and placebo spray (0.0) groups. CONCLUSION: Antimicrobial sprays were shown to be effective among institutionalised elders. Chlorine dioxide spray showed equivalent effects on dental plaque and gingival bleeding relative to the chlorhexidine spray over a 6-month period.

Lidocaine spray versus viscous lidocaine solution for pharyngeal local anesthesia in upper gastrointestinal endoscopy: Systematic review and meta-analysis.

OBJECTIVES: There are two major methods for local anesthesia by lidocaine before upper gastrointestinal endoscopy: simple spray and viscous solution. We aimed to assess the efficacy and safety by meta-analysis between these two methods. METHODS: We searched PubMed, EMBASE, the Cochrane Central Register of Controlled Trials, World Health Organization International Clinical Trials Registry Platform, and ClinicalTrials.gov databases through October 2019 to perform meta-analyses using random-effects models. The primary outcomes were participants' pain/discomfort, satisfaction, and anaphylactic shock. Three reviewers independently searched for articles, extracted data, and assessed the risk of bias. We evaluated the certainty of evidence based on the Grading of Recommendations, Assessment, Development, and Evaluation approach. This study was registered in PROSPERO (CRD42020155611). RESULTS: We included seven randomized controlled trials (2667 participants). The participants' pain/discomfort may be similar between the lidocaine spray and viscous solution [standardized mean difference 0.03, 95% confidence intervals (CI) -0.37 to 0.42; I2  = 93%; low certainty of evidence]. The lidocaine spray probably increased participants' satisfaction compared with the viscous solution (relative risk 1.22; 95% CI, 1.02 to 1.47; I2  = 47%; moderate certainty of evidence). No anaphylactic shock occurred in four studies (low certainty of evidence). Four studies had high risks of selection bias. CONCLUSION: The use of lidocaine spray for local anesthesia provided better satisfaction scores than the viscous solution, and both methods have the same effect with regards to the control of discomfort and pain. Further studies in large multicenter randomized controlled trials with a pre-registration protocol are needed.

[Effect of circular RNA hsa_circ_0008898 on oral squamous cell carcinoma and its mechanism].

Objective: To investigate the effect and molecular mechanism of hsa_circ_0008898 on the cell proliferation, migration, invasion and tumor formation of oral squamous cell carcinoma (OSCC). Methods: Quantitative real-time PCR (qPCR) and Western blotting were used to detect the expression of hsa_circ_0008898, miR-197-5p and ras homolog gene family member A (RHOA) in OSCC tissues, adjacent tissues, OSCC cells and human normal oral keratinocytes (NOK). CAL27 and SCC-25 cells were transfected with si-hsa_circ_0008898#1 (knockdown group 1), si-hsa_circ_0008898#2 (knockdown group 2), hsa_circ_0008898 (circ overexpression group) and blank plasmid (circ blank group), respectively. Then miR-197-5p inhibitor (inhibition group) and blank plasmid (inhibition control group) were transfected into hsa_circ_0008898 knockdown cells (knockdown group 1). CAL27 and SCC-25 cells were transfected with miR-197-5p mimics (miR overexpression group) and blank plasmid (miR blank group), and then transfected hsa_circ_0008898 vector (co-transfection group 1), RHOA vector (co-transfection group 2) and blank plasmid (co-transfection control group) in cells overexpressing miR-197-5p. Cell counting kit 8 (CCK-8), colony formation, Transwell and scratch test were used to detect cell proliferation, cloning ability, cell cycle distribution, cell invasion and migration ability. Ten nude mice were equally divided into two groups, with 5 mice in each group. SCC-25 cells transfected with blank plasmid (control group) and SCC-25 cells transfected with sh-hsa_circ_0008898 (knockout group) were subcutaneously injected into the armpit. The volume and mass of the tumor were measured. Results: The expressions of hsa_circ_0008898 (2.89±0.72) and RHOA (2.62±0.21) in OSCC tissues were significantly higher than those in para-carcinoma tissues (1.00±0.48, 1.00±0.11, respectively), while the expression of miR-197-5p in OSCC tissues （0.46±0.24） was significantly lower than that in para-carcinoma tissues (1.00±0.42) (P<0.05). Compared with NOK, the expression of hsa_circ_0008898 and RHOA in CAL27 and SCC-25 cells increased significantly, while the expression of miR-197-5p decreased (P<0.05). Compared with circ blank group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 cells in knockdown group 1 and group 2 were significantly decreased, while the proportion of cells in G1 phase was significantly increased (P<0.05). Compared with inhibition control group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 in inhibition group were significantly increased, while the proportion of cells in G1 phase was significantly decreased in inhibition group (P<0.05). Compared with miR blank group, the cell viability, colony formation, scratch healing rate and invasive cell number of CAL27 and SCC-25 in miR overexpression group were significantly decreased, while the proportion of cells in G1 phase was significantly increased in miR overexpression group (P<0.05). Compared with co-transfection control group, the cell viability, colony formation, migration area and invasive cell number of CAL27 and SCC-25 in co-transfection group2 were significantly increased, while the proportion of cells in G1 phase was significantly decreased in co-transfection group 2 (P<0.05). The volume and mass of transplanted tumor in knockout group ï¼»(660.4±67.8) mm(3 )and (0.60±0.06) g, respectivelyï¼½ were significantly lower than those in control group ï¼»(1 210.4±198.9) mm(3) and (1.00±0.12) g, respectivelyï¼½. Conclusions: Knockdown of hsa_circ_0008898 inhibited OSCC cells proliferation, cloning, migration and invasion and induced cell cycle arrest in vitro by regulating the miR-197-5p/RHOA. Additionally, Knockdown of hsa_circ_0008898 also inhibited tumor formation of OSCC cells in vivo.

Prognostic value of serum MIR4435-2HG in oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To investigate the value of serum MIR4435-2HG level in the diagnosis and prognosis of oral squamous cell carcinoma.@*Methods@#This study was a retrospective case-control study. Five hundred and eighteen samples of oral squamous carcinoma of patients with head and neck squamous cell carcinoma in the cancer genome atlas project (TCGA) database, with long noncoding RNA MIR4435-2HG expression. The median was the boundary, and the patients were divided into high expression group and low expression group, and the 5-year disease-free survival rate and overall survival rate of the two groups were compared. Serum samples from 82 patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Affiliated Stomatology Hospital of Huzhou Univerisity from January 2012 to January 2015 were enrolled to verify the prognostic value of MIR4435-2HG. Bioinformatics is used to predict the biological processes involved in MIR4435-2HG. Use the SPSS 23.0 to set the optimal diagnostic and prognostic cutoff for the MIR4435-2HG.@*Results@#A total of 518 oral squamous carcinoma patients in the TCGA database showed that the 5-year overall survival rate of the MIR4435-2HG high expression group [43.2% (112/259)] was significanthy lower than that of the MIR4435-2HG low expression group [51.7% (134/259)] (P<0.05). The disease-free survival rate of the MIR4435-2HG high expression group [56.8% (147/259)] was significantly lower than that of the MIR4435-2HG low expression group [64.1% (166/259)] (P<0.05). The results of the validation of 82 patients with oral squamous cell carcinoma suggested that the 3-year overall survival rate of the MIR4435-2HG high expression group [40.0% (8/20)] was significantly lower than the MIR4435-2HG low expression group [80.6 % (50/62)] (P<0.05). The clinical and pathological data of serum MIR4435-2HG high expression group and serum MIR4435-2HG low expression group were compared. The results showed that there was no significant difference in gender, age, tumor location and TNM staging between the two groups (P>0.05). The lymph node metastasis rate of the MIR4435-2HG high expression group was significantly higher than that of the low expression group [12.9% (8/62)] (P<0.05). The histological grade of the high expression group [80.0 % (16/20)] was significantly lower than that of the low expression group [24.2 % (15/62)] (χ2=20.030, P<0.05). The results of bioinformatics analysis indicated that the biological functions of MIR4435-2HG target gene were mainly enriched in protein metabolism, processing of rRNA in nucleolus and cytoplasm, SEMA4D induced cell migration process, and mitochondrial translation process.@*Conclusions@#Serum MIR4435-2HG can be used as a potential prognostic marker for oral squamous cell carcinoma.

[Effect of circular RNA hsa_circ_0063772 on proliferation, migration and invasion of oral squamous cell carcinoma cells].

Objective: To investigate the effects of circular RNA hsa_circ_0063772 on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells. Methods: Thirty-three patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital from February 2017 to December 2018 were enrolled in this study. Real-time quantative polymerase chain reaction was used to detect the expression level of circular RNA hsa_circ_0063772 in OSCC and corresponding adjacent tissues, OSCC cell lines and human keratinocytes. The expression level of hsa_circ_0063772 was overexpressed in SCC15 and CAL27 cells by using lentivirus, and the effects of this gene on proliferation, migration and invasion of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay, Western blotting and nude mice tumor formation assay. Results: The expression of circular RNA hsa_circ_0063772 in OSCC tissues (9.38±0.34) was lower than that in adjacent tissues (11.30±0.31) (t=5.20, P<0.001), and the expression in OSCC cells (SCC15: 0.12±0.01; SCC25: 0.18±0.02; SCC9: 0.21±0.01; CAL27: 0.13±0.01) was significantly lower than that in human keratinocytes (1.02±0.02) (t(SCC15)=41.91, t(SCC25)=29.21, t(SCC9)=35.16, t(CAL27)=41.86, P<0.001). Overexpression of hsa_circ_0063772 in SCC15 and CAL27 cells can affect tumor cell proliferation, cell counting assay showed that tumor cell proliferation ability in high expression group (SCC15: 0.76±0.01; CAL27: 0.74±0.02) were lower than empty group (SCC15∶1.22±0.04; CAL27: 0.99±0.03; t(SCC15)=12.58, t(CAL27)=6.97; P<0.05). Transwell migration experiment showed the number of migrated cell in high expression group (SCC15∶148.00±14.57; CAL27: 243.00±13.00) were less than empty group (SCC15: 580.30±42.91; CAL27: 424.70±18.66, P<0.01); Transwell invasion assay showed the number of invased cell in high expression group (SCC15: 123.70±6.98; CAL27: 326.00±17.01) were less than empty group (SCC15: 517.70±9.96; CAL27: 454.30±8.09, P<0.01). The results of tumor formation in nude mice showed that the tumor volume and mass of the overexpressed group [(306.40±16.51) mm(3); (289.40±11.44) mg] were lower than that of the empty group [(582.60±32.51) mm(3), t=7.58, P<0.05; (599.60±21.27) mg, t=7.58, P<0.001]. Conclusions: Compared with adjacent tissues, hsa_circ_0063772 is lowly expressed in oral squamous cell carcinoma. High expression of hsa_circ_0063772 can inhibit the proliferation, migration and invasion of OSCC cells.

Effect of Oral Spray on Dental Plaque Bacteria and Oral Epithelial Cells / 치위생과학회지

BACKGROUND: Good oral health is important for systemic body health and quality of life. Spray oral cleansers are increasingly preferred because of their convenience of carrying and the ease of oral hygiene management. In addition, many kinds of oral cleanser products containing various ingredients with antibacterial, washing, and moisturizing effects are being manufactured. However, concerns about the safety and side effects of oral sprays are increasing, and there is very little information regarding the use and care of oral sprays is available to consumers. This study aimed to investigate the effects of oral spray on oral bacteria and tissue to elucidate the factors that need to be considered when using oral sprays. METHODS: The effects of oral spray on the growth of dental plaque bacteria was assessed using disk diffusion assays. Cytotoxicity and morphological changes in oral epithelial cells were observed by microscopy. The effects of oral spray on dental plaque growth were also confirmed on specimens from permanent incisors of bovines by Coomassie staining. RESULTS: The pH of spray products, such as Perioe Dental Cooling, Cool Sense, and Dentrix, were 3.65, 3.61, and 6.15, respectively. All tested spray products showed strong toxicity to dental plaque bacteria and oral epithelial cells. Compared with those on the control, dental plaque bacteria deposits on the enamel surface increased following the use of oral spray. CONCLUSION: Three types of oral spray, namely Perioe Dental Cooling, Cool Sense, and Dentrix, strongly inhibited the growth of dental plaque bacteria and oral epithelial cells. The oral spray ingredient enhanced dental plaque growth on the enamel surface. Users should be informed of precautions when using oral sprays and the need for oral hygiene after its use.

Effect of circular RNA hsa_circ_0063772 on proliferation, migration and invasion of oral squamous cell carcinoma cells / 中华口腔医学杂志

Objective@#To investigate the effects of circular RNA hsa_circ_0063772 on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells.@*Methods@#Thirty-three patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital from February 2017 to December 2018 were enrolled in this study. Real-time quantative polymerase chain reaction was used to detect the expression level of circular RNA hsa_circ_0063772 in OSCC and corresponding adjacent tissues, OSCC cell lines and human keratinocytes. The expression level of hsa_circ_0063772 was overexpressed in SCC15 and CAL27 cells by using lentivirus, and the effects of this gene on proliferation, migration and invasion of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay, Western blotting and nude mice tumor formation assay.@*Results@#The expression of circular RNA hsa_circ_0063772 in OSCC tissues (9.38±0.34) was lower than that in adjacent tissues (11.30±0.31) (t=5.20, P<0.001), and the expression in OSCC cells (SCC15: 0.12±0.01; SCC25: 0.18±0.02; SCC9: 0.21±0.01; CAL27: 0.13±0.01) was significantly lower than that in human keratinocytes (1.02±0.02) (tSCC15=41.91, tSCC25=29.21, tSCC9=35.16, tCAL27=41.86, P<0.001). Overexpression of hsa_circ_0063772 in SCC15 and CAL27 cells can affect tumor cell proliferation, cell counting assay showed that tumor cell proliferation ability in high expression group (SCC15: 0.76±0.01; CAL27: 0.74±0.02) were lower than empty group (SCC15∶1.22±0.04; CAL27: 0.99±0.03; tSCC15=12.58, tCAL27=6.97; P<0.05). Transwell migration experiment showed the number of migrated cell in high expression group (SCC15∶148.00±14.57; CAL27: 243.00±13.00) were less than empty group (SCC15: 580.30±42.91; CAL27: 424.70±18.66, P<0.01); Transwell invasion assay showed the number of invased cell in high expression group (SCC15: 123.70±6.98; CAL27: 326.00±17.01) were less than empty group (SCC15: 517.70±9.96; CAL27: 454.30±8.09, P<0.01). The results of tumor formation in nude mice showed that the tumor volume and mass of the overexpressed group [(306.40±16.51) mm3; (289.40±11.44) mg] were lower than that of the empty group [(582.60±32.51) mm3, t=7.58, P<0.05; (599.60±21.27) mg, t=7.58, P<0.001].@*Conclusions@#Compared with adjacent tissues, hsa_circ_0063772 is lowly expressed in oral squamous cell carcinoma. High expression of hsa_circ_0063772 can inhibit the proliferation, migration and invasion of OSCC cells.

[Relationship between expression level of sex-determining region Y box 9 and metastasis of oral squamous cell carcinoma].

Objective: To investigate the effects of sex-detemining region Y box9 (SOX9) expression levels on the proliferation, migration and metastasis in oral squamous cell carcinoma (OSCC). Methods: A total of 74 OSCC pathological specimens were collected from Shanghai OSCC Tissue and Biological Informations Bank, and clinicopathological information of these specimens were collected. Immunohistochemistry assay was used to examine the expression levels of SOX9 in OSCC and to analyze their relationship with clinicopathological features. Cell counting kit-8 assay and cloning formation was used to observe the relationship between the expression levels of SOX9 and the proliferation of OSCC. Transwell experiment and scratch test were used to detect the difference of the ability of OSCC in cell lines with different expression levels of SOX9. Results: The risk of lymph node metastasis in patients with high expression of SOX9 was significantly increased (P=0.010). In the Transwell experiment, the number of HN6 cells (671.0±57.4, P=0.000) migrated to the lower chamber more than that of CAL27 cells (172.0±13.9). In the scratch experiment, HN6 cells [0 h: (93.7±2.1) µm; 6 h: (56.7±2.5) µm; 12 h: (29.7±3.1) µm] migrated faster than CAL27 [0 h: (93.7±1.5) µm; 6 h: (78.0±2.0) µm; 12 h: (42.0±3.0) µm](P<0.05). The migration ability of the cell line (HN6) with high-expression of SOX9 was significantly higher than that in cell line (CAL27) with low-expression SOX9 (P<0.05). The expression levels of SOX9 in OSCC were no significant on cell proliferation (P>0.05). Conclusions: High expression of SOX9 can promote the migration and lymph node metastasis of OSCC. SOX9 is a candidate gene target for the diagnosis and intervention of lymph node metastasis in OSCC.

[Effect of long non-coding RNA highly upregulated in liver cancer on the biological behavior of oral squamous cell carcinoma].

Objective: To investigate the effect of long non-coding RNA highly upregulated in liver cancer (lncRNA HULC) on the biological behavior of oral squamous cell carcinoma (OSCC) cell lines. Methods: A total of thirty patients with oral squamous cell carcinoma at Peking University Shenzhen Hospital from March 2017 to March 2018 were enrolled in this study. OSCC and adjacent tissues were extracted during tumor extensive resection. Quantitative real-time PCR (qPCR) was used to detect the expression levels of lncRNA HULC in OSCC and paracancerous tissues and OSCC cell lines. SCC15 and SCC25 cells were transfected with siRNA, and the effects of the gene on the biological behavior of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay and Western blotting. Results: The expression of lncRNA HULC in OSCC tissues (10.98±0.31, n=30) was significantly higher than that in paracancerous tissues (8.39±0.31, n=30) (t=5.93, P<0.001), the expression of lncRNA HULC in OSCC cells (SCC15: 28.58±2.74; SCC25: 16.56±0.87; SCC9: 11.18±1.32; CAL27: 13.92±0.99, n=5) was significantly higher than that in human keratinocytes (1.01±0.00, n=5) (t(SCC15)=10.08, t(SCC25)=17.96, t(SCC9)=7.71, t(CAL27)=13.09, P<0.001). Down-regulation of lncRNA HULC in SCC15 and SCC25 cells can inhibit the proliferation, migration and invasion of tumor cells and promote tumor cell apoptosis. Conclusions: lncRNA HULC is highly expressed in OSCC and can enhance the proliferation, migration and invasion of cancer cells and inhibit tumor cell apoptosis.

[Circular RNA circHIPK3 acts as the sponge of microRNA-124 to promote human oral squamous cell carcinoma cells proliferation].

Objective: To explore the expression and clinical significance of circular RNA circHIPK3 in oral squamous cell carcinoma (OSCC), analyze the effect of circHIPK3 on the proliferation of OSCC cells. Methods: The expression of circHIPK3 in OSCC tissues, adjacent non-cancerous tissues and OSCC cell lines were detected by quantitative real-time polymerase chain reaction (qPCR). The correlations between the expression of circHIPK3 in OSCC tissues and the clinicopathological features were analyzed as well. circHIPK3-specific siRNA si-circHIPK3 and negative control siRNA si-NC were designed and synthesised and used to transfect CAL27 and SCC15 cells respectively. The proliferation capacity of CAL27 and SCC15 cells after transfection with si-circHIPK3 was detected by CCK-8 assay. The expression of miR-124 in OSCC was detected by qPCR, and the correlation between expression of circHIPK3 and the expression of miR-124 was analyzed. Using qPCR to detect the expression of miR-124 in CAL27 and SCC15 cells after transfection with si-circHIPK3 and si-NC respectively. Furthermore, using CCK-8 assay to detect the proliferation capacity of CAL27 and SCC15 cells after transfection with si-NC, si-circHIPK3, miR-124 mimic, si-circHIPK3+miR-124 inhibitor. Results: The expression of circHIPK3 in OSCC tissues [2.23 (1.86, 3.00)] was significantly higher than that of the adjacent non-cancerous tissues [1.05 (0.85, 1.26)] (U=1 094, P=0.000). The expression of circHIPK3 in CAL27 (3.02±0.51) and SCC15 cells (3.16±0.75) was higher than those of human normal oral keratinocytes (hNOK) (1.26±0.30) (P=0.000). The expression of circHIPK3 was found to be closely associated with TNM stage (P<0.05) and tumor grades (P<0.05). Knockdown of circHIPK3 can inhibit proliferation of CAL27 and SCC15 cells (P<0.05). The expression of miR-124 in OSCC tissues (0.61±0.35) was significantly lower than that in adjacent non-cancerous tissues (1.13+0.39) (t=-5.36, P<0.05). Correlation analysis showed that the expression of circHIPK3 in OSCC was negatively correlated with the expression of miR-124 (r=-0.767, P<0.001). Moreover, down-regulation of miR-124 rescued the phenotype induced by knockdown of circHIPK3. Conclusions: The expression of circHIPK3 in OSCC was increased, and silencing of circHIPK3 expression can inhibit the proliferation of OSCC cells. Our results suggest that circHIPK3 may play a key role in the occurrence and development process of OSCC through the regulation of miR-124 expression.

[Expression and potential clinical significance of cytidine triphosphate: phosphocholine cytidylyltransferase-α in oral squamous cell carcinoma].

Objective: To examine the expression and potential clinical significance of CCT (cytidine triphosphate: phosphocholine cytidylyltransferase)-α in oral squamous cell carcinoma (OSCC). Methods: Fifty-eight OSCC and paired adjacent non-malignant epithelia samples (between May 2016 and July 2016) were obtained from dental center, Second Xiangya Hospital, Central South University. CCT-α expression was examined by immunohistochemistry. The relationship between CCT-α and clinicopathological features of OSCC patients was analyzed. Quantitative real-time PCR and Western blot were performed to measure the expression of CCT-α mRNA and protein level in several OSCC cell line and two normal oral epithelial cell line. Results: Immunohistochemistry showed that CCT-α positive staining was found in cell nuclear of OSCC cells and adjacent epithelial cells. CCT-α was positively expressed in OSCC, which was significantly higher than that adjacent to carcinoma tissues (P=0.000). The expression of CCT-α in oral squamous cell carcinoma was correlated with smoking, alcohol consumption, tumor size, differentiation degree and lymph node metastasis. The expression level of CCT-α protein was significantly increased in patients with a history of smoking and alcohol consumption (P=0.001, P=0.004). With the increase of tumor diameter, the expression of CCT-α protein was significantly increased (P=0.005). According to histopathological grade, the lower the degree of tumor differentiation, the higher the expression level of CCT-α protein (P=0.000). The expression of CCT-α protein was significantly higher in patients with lymph node metastasis compared with no lymph node metastasis (P=0.000). Quantitative real-time PCR results showed the CCT-α mRNA expression level was significantly higher in OSCC cells than that in normal oral epithelial cells (P=0.016). The protein expression level of CCT-α was significantly higher in OSCC cells than that in normal oral epithelial cells. Conclusions: CCT-α may play a critical role in the carcinogenesis and development of OSCC.

Circular RNA circHIPK3 acts as the sponge of microRNA-124 to promote human oral squamous cell carcinoma cells proliferation / 中华口腔医学杂志

Objective@#To explore the expression and clinical significance of circular RNA circHIPK3 in oral squamous cell carcinoma (OSCC), analyze the effect of circHIPK3 on the proliferation of OSCC cells.@*Methods@#The expression of circHIPK3 in OSCC tissues, adjacent non-cancerous tissues and OSCC cell lines were detected by quantitative real-time polymerase chain reaction (qPCR). The correlations between the expression of circHIPK3 in OSCC tissues and the clinicopathological features were analyzed as well. circHIPK3-specific siRNA si-circHIPK3 and negative control siRNA si-NC were designed and synthesised and used to transfect CAL27 and SCC15 cells respectively. The proliferation capacity of CAL27 and SCC15 cells after transfection with si-circHIPK3 was detected by CCK-8 assay. The expression of miR-124 in OSCC was detected by qPCR, and the correlation between expression of circHIPK3 and the expression of miR-124 was analyzed. Using qPCR to detect the expression of miR-124 in CAL27 and SCC15 cells after transfection with si-circHIPK3 and si-NC respectively. Furthermore, using CCK-8 assay to detect the proliferation capacity of CAL27 and SCC15 cells after transfection with si-NC, si-circHIPK3, miR-124 mimic, si-circHIPK3+miR-124 inhibitor.@*Results@#The expression of circHIPK3 in OSCC tissues [2.23 (1.86, 3.00)] was significantly higher than that of the adjacent non-cancerous tissues [1.05 (0.85, 1.26)] (U=1 094, P=0.000). The expression of circHIPK3 in CAL27 (3.02±0.51) and SCC15 cells (3.16±0.75) was higher than those of human normal oral keratinocytes (hNOK) (1.26±0.30) (P=0.000). The expression of circHIPK3 was found to be closely associated with TNM stage (P<0.05) and tumor grades (P<0.05). Knockdown of circHIPK3 can inhibit proliferation of CAL27 and SCC15 cells (P<0.05). The expression of miR-124 in OSCC tissues (0.61±0.35) was significantly lower than that in adjacent non-cancerous tissues (1.13+0.39) (t=-5.36, P<0.05). Correlation analysis showed that the expression of circHIPK3 in OSCC was negatively correlated with the expression of miR-124 (r=-0.767, P<0.001). Moreover, down-regulation of miR-124 rescued the phenotype induced by knockdown of circHIPK3.@*Conclusions@#The expression of circHIPK3 in OSCC was increased, and silencing of circHIPK3 expression can inhibit the proliferation of OSCC cells. Our results suggest that circHIPK3 may play a key role in the occurrence and development process of OSCC through the regulation of miR-124 expression.

Expression and potential clinical significance of cytidine triphosphate: phosphocholine cytidylyltransferase-α in oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To examine the expression and potential clinical significance of CCT (cytidine triphosphate: phosphocholine cytidylyltransferase)-α in oral squamous cell carcinoma (OSCC).@*Methods@#Fifty-eight OSCC and paired adjacent non-malignant epithelia samples (between May 2016 and July 2016) were obtained from dental center, Second Xiangya Hospital, Central South University. CCT-α expression was examined by immunohistochemistry. The relationship between CCT-α and clinicopathological features of OSCC patients was analyzed. Quantitative real-time PCR and Western blot were performed to measure the expression of CCT-α mRNA and protein level in several OSCC cell line and two normal oral epithelial cell line.@*Results@#Immunohistochemistry showed that CCT-α positive staining was found in cell nuclear of OSCC cells and adjacent epithelial cells. CCT-α was positively expressed in OSCC, which was significantly higher than that adjacent to carcinoma tissues (P=0.000). The expression of CCT-α in oral squamous cell carcinoma was correlated with smoking, alcohol consumption, tumor size, differentiation degree and lymph node metastasis. The expression level of CCT-α protein was significantly increased in patients with a history of smoking and alcohol consumption (P=0.001, P=0.004). With the increase of tumor diameter, the expression of CCT-α protein was significantly increased (P=0.005). According to histopathological grade, the lower the degree of tumor differentiation, the higher the expression level of CCT-α protein (P=0.000). The expression of CCT-α protein was significantly higher in patients with lymph node metastasis compared with no lymph node metastasis (P=0.000). Quantitative real-time PCR results showed the CCT-α mRNA expression level was significantly higher in OSCC cells than that in normal oral epithelial cells (P=0.016). The protein expression level of CCT-α was significantly higher in OSCC cells than that in normal oral epithelial cells.@*Conclusions@#CCT-α may play a critical role in the carcinogenesis and development of OSCC.

Effect of long non-coding RNA highly upregulated in liver cancer on the biological behavior of oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To investigate the effect of long non-coding RNA highly upregulated in liver cancer (lncRNA HULC) on the biological behavior of oral squamous cell carcinoma (OSCC) cell lines.@*Methods@#A total of thirty patients with oral squamous cell carcinoma at Peking University Shenzhen Hospital from March 2017 to March 2018 were enrolled in this study. OSCC and adjacent tissues were extracted during tumor extensive resection. Quantitative real-time PCR (qPCR) was used to detect the expression levels of lncRNA HULC in OSCC and paracancerous tissues and OSCC cell lines. SCC15 and SCC25 cells were transfected with siRNA, and the effects of the gene on the biological behavior of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay and Western blotting.@*Results@#The expression of lncRNA HULC in OSCC tissues (10.98±0.31, n=30) was significantly higher than that in paracancerous tissues (8.39±0.31, n=30) (t=5.93, P<0.001), the expression of lncRNA HULC in OSCC cells (SCC15: 28.58±2.74; SCC25: 16.56±0.87; SCC9: 11.18±1.32; CAL27: 13.92±0.99, n=5) was significantly higher than that in human keratinocytes (1.01±0.00, n=5) (tSCC15=10.08, tSCC25=17.96, tSCC9=7.71, tCAL27=13.09, P<0.001). Down-regulation of lncRNA HULC in SCC15 and SCC25 cells can inhibit the proliferation, migration and invasion of tumor cells and promote tumor cell apoptosis.@*Conclusions@#lncRNA HULC is highly expressed in OSCC and can enhance the proliferation, migration and invasion of cancer cells and inhibit tumor cell apoptosis.

Relationship between expression level of sex-determining region Y box 9 and metastasis of oral squamous cell carcinoma / 中华口腔医学杂志

Objective@#To investigate the effects of sex-detemining region Y box9 (SOX9) expression levels on the proliferation, migration and metastasis in oral squamous cell carcinoma (OSCC).@*Methods@#A total of 74 OSCC pathological specimens were collected from Shanghai OSCC Tissue and Biological Informations Bank, and clinicopathological information of these specimens were collected. Immunohistochemistry assay was used to examine the expression levels of SOX9 in OSCC and to analyze their relationship with clinicopathological features. Cell counting kit-8 assay and cloning formation was used to observe the relationship between the expression levels of SOX9 and the proliferation of OSCC. Transwell experiment and scratch test were used to detect the difference of the ability of OSCC in cell lines with different expression levels of SOX9.@*Results@#The risk of lymph node metastasis in patients with high expression of SOX9 was significantly increased (P=0.010). In the Transwell experiment, the number of HN6 cells (671.0±57.4, P=0.000) migrated to the lower chamber more than that of CAL27 cells (172.0±13.9). In the scratch experiment, HN6 cells [0 h: (93.7±2.1) μm; 6 h: (56.7±2.5) μm; 12 h: (29.7±3.1) μm] migrated faster than CAL27 [0 h: (93.7±1.5) μm; 6 h: (78.0±2.0) μm; 12 h: (42.0±3.0) μm](P<0.05). The migration ability of the cell line (HN6) with high-expression of SOX9 was significantly higher than that in cell line (CAL27) with low-expression SOX9 (P<0.05). The expression levels of SOX9 in OSCC were no significant on cell proliferation (P>0.05).@*Conclusions@#High expression of SOX9 can promote the migration and lymph node metastasis of OSCC. SOX9 is a candidate gene target for the diagnosis and intervention of lymph node metastasis in OSCC.

[Evaluation and influencing factors of surgical margin status of oral squamous cell carcinoma].

Surgical resection with adequate margins is an essential component of the treatment for patients with oral squamous cell carcinoma (OSCC). A distance of 5 mm or more between healthy tissue to the tumor front is generally accepted as a safe margin. It is very important for surgeons to precisely evaluate the resection area of tumor both pre- and intra-operatively and try to achieve a safe margin, which will result in a decreased risk of local recurrence. The relationship of surgical margin status to patients' prognosis, and factors which will affect surgical margin distance demand are discussed in this paper. We recommend that adequate margins evaluation should take consideration of many factors such as anatomical location, depth of tumor invasion, pattern of tumor invasion, mucosal dysplasia grade and so on. With the development of molecular biology, surgical margin study at molecular level can give us a new strategy to evaluate its adequacy.

Evaluation and influencing factors of surgical margin status of oral squamous cell carcinoma / 中华口腔医学杂志

Surgical resection with adequate margins is an essential component of the treatment for patients with oral squamous cell carcinoma (OSCC). A distance of 5 mm or more between healthy tissue to the tumor front is generally accepted as a safe margin. It is very important for surgeons to precisely evaluate the resection area of tumor both pre- and intra-operatively and try to achieve a safe margin, which will result in a decreased risk of local recurrence. The relationship of surgical margin status to patients' prognosis, and factors which will affect surgical margin distance demand are discussed in this paper. We recommend that adequate margins evaluation should take consideration of many factors such as anatomical location, depth of tumor invasion, pattern of tumor invasion, mucosal dysplasia grade and so on. With the development of molecular biology, surgical margin study at molecular level can give us a new strategy to evaluate its adequacy.

Effect of triptolide on human oral cancer cell proliferation and PTEN gene mRNA expression in oral cancer / 中华口腔医学杂志

Objective@#To investigate the effect of triptolide on human oral cancer cell (HB) proliferation and phosphates and tensin homologue deleted on chromosome ten gene (PTEN) mRNA expression in oral cancer.@*Methods@#The cancer cells were cultured in the medium containing triptolide of different concentrations for 24, 48 and 72 h. Methyl thiazolyl tetrazolium (MTT) method was used to test the rate of growth inhibition of cancer cells, flow cytometer to detect the change of cell cycle and reveres transcription-PCR (RT-PCR) to examine the expression of PTEN mRNA. The expression of PTEN protein was examined by Western blotting.@*Results@#The rate of growth inhibition was (26.92 ± 0.14)%, (38.67 ± 0.11)%, (72.62 ± 0.89)% and (90.42 ± 0.28)%, respectively. The corresponding expression of PTEN mRNA was (3.59±0.21)%, (5.27±0.40)%, (7.18±0.44)% and (9.16±0.50)%, respectively and the corresponding A value of PTEN protein was 0.135±0.007, 0.410±0.020, 0.447±0.017 and 0.884±0.066, respectively. The proportion of G1 phase cells increased from (58.78±0.98)% to (84.13±0.47)%, but the proportion of S phase cells decreased from (25.40±0.43)% to (9.41±0.73)%.@*Conclusions@#The triptolide not only had inhibitory effect on the HB proliferation, but also affected the cell cycle.

Constructing an interaction network of differential genes of oral squamous cell carcinoma with RACK1 as a core / 中国组织工程研究

BACKGROUND:RACK1 is strongly associated with the occurrence and development of oral squamous cel carcinoma. However, the occurrence and development of tumor do not depend on a gene or protein, but a long-term complex process of a network structure of multiple genes and multiple molecules, multi-step, multi-stage joint action. Synergism between tumor genes promotes the formation and development of tumor cel s. Therefore, we cannot limit on a single gene or protein to discover the action mechanism of oral squamous cel carcinoma, but should pay attention on signaling network path related to differential protein or gene, investigate the alterations in related protein or gene expression in the whole signaling pathway, and analyze the action mechanism of the interaction of these molecules. OBJECTIVE:To screen differential genes related to oral squamous cel carcinoma, construct an interaction network through bioinformatics using STRING database, and provide clues for future tests. METHODS:In accordance with our previous classic proteomics results and microarray results of oral squamous cel carcinoma, genes with consistent expression and big differences were selected as differential genes. The differential genes were inputted into the database of STRING to find the possible relationship among the protein subunits and to construct network structure of their interaction. RESULTS AND CONCLUSION:The 19 differential proteins of oral squamous cel carcinoma construct a complicated net work, and the differential proteins interact through these networks. GNB2L1-encoded RACK1 is a node protein and interacts with other differential proteins via WD40 repeated protein (number COG2319) andβ-G protein subunit (number KOG0279). WD40 repeated protein (number COG2319) interacts with 5 differential proteins directly and constructs 10 interacting pathways.β-G protein subunit (number KOG0279) interacts with 8 differential proteins directly, which has 11 interacting pathways. We make a network structure picture based on the interaction of these 19 differential genes by the analysis of the STRING database. The results show that the two subunits of RACK1 protein have direct interaction with 8 differential proteins and have 18 interaction pathways on the picture. As a result, RACK1 is the core protein of the network, suggesting RACK1 is the key node protein in oral squamous cel carcinoma.