Identification of prognostic m6A modification patterns and score system in melanoma patients.

N6-methyladenosine (m6A) is the most common modification on RNAs and LncRNAs. It plays an important role in cancer stem cell differentiation, T cell differentiation, and immune homeostasis. In this study, we explored the potential roles of m6A modification of RNA in melanoma and investigated the immune cell infiltration in tumor microenvironment in diverse m6Aclusters and different m6Ascore groups. A consensus clustering algorithm determined m6A modification patterns based on 14 m6A regulators, and further explored the biological functions and the connection with TME. An m6A-related gene signature (m6Ascore) was constructed based on m6A-related genes using principal component analysis. Three m6A modification patterns were identified based on 14 m6A regulators, named as m6Aclusters A-C. The prognosis of m6Acluster A was more favorable than m6Aclusters B and C, and it was more closely associated with immune regulation. To quantify the m6A modification patterns of individual tumor, an m6Ascore was constructed, and patients were classified into high and low m6Ascore groups. The low m6Ascore group, which had a favorable prognosis, was more relevant to immunology. The expression of PD-L1 was higher and the immunophenoscore (IPS) revealed stronger response to immunotherapy in the low m6Ascore group. This study identified 3 m6A modification patterns with different immune characteristics and constructed an m6Ascore system to predict prognosis and immunogenicity of patients, which is conducive to clinical prognosis judgment and individual treatment.

Periodontitis aggravates COPD through the activation of γδ T cell and M2 macrophage.

Chronic obstructive pulmonary disease (COPD) is a chronic systemic inflammatory disease with high morbidity and mortality. Periodontitis exacerbates COPD progression; however, the immune mechanisms by which periodontitis affects COPD remain unclear. Here, by constructing periodontitis and COPD mouse models, we demonstrated that periodontitis and COPD could mutually aggravate disease progression. For the first time, we found that the progression was associated with the activation of γδ T cells and M2 macrophages, and M2 polarization of macrophages was affected by γδ T cells activation. In the lung tissues of COPD with periodontitis, the activation of γδ T cells finally led to the increase of IL 17 and IFN γ expression and M2 macrophage polarization. Furthermore, we found that the periodontitis-associated bacteria Porphyromonas gingivalis (P. gingivalis) promoted the activation of γδ T cells and M2 macrophages ex vivo. The data from clinical bronchoalveolar lavage fluid (BALF) samples were consistent with the in vivo and ex vivo experiments. For the first time, our results identified the crucial role of γδ T-M2 immune mechanism in mediating periodontitis-promoted COPD progression. Therefore, targeting at periodontitis treatment and the γδ T-M2 immune mechanism might provide a new practical strategy for COPD prevention or control.IMPORTANCEPeriodontitis exacerbates chronic obstructive pulmonary disease (COPD) progression. For the first time, the current study identified that the impact of periodontitis on COPD progression was associated with the activation of γδ T cells and M2 macrophages and that M2 polarization of macrophages was affected by γδ T cells activation. The results indicated that targeting at periodontitis treatment and the γδ T-M2 immune mechanism might provide a new practical strategy for COPD prevention or control.

Validity and Reliability of the Metacognitions Questionnaire-30 (MCQ-30) among Chinese Adolescents.

This study aimed to examine the psychometric properties of the Metacognitions Questionnaire-30 (MCQ-30) in a sample of Chinese adolescents (1382 boys, 1445 girls) aged 11 to 18 years. Confirmatory factor analysis was performed to assess factor structure, as well as, measurement invariance across demographic groups and clinical symptoms. The results of confirmatory factor analyses supported the original five-factor model. Configural, metric and scalar invariance of the five-factor model were also supported by gender, age, ethnicity, residence, parental education level, depression and anxiety status. Furthermore, all five subscales demonstrated good internal consistency (Cronbach alphas > 0.75) and test-retest reliability (intra-class correlation coefficients > 0.45). Finally, the five factors were positively related to symptoms of depression, anxiety, and irritability and negatively related to positive childhood experiences and life satisfaction, indicating excellent validity. The findings provide initial evidence that the MCQ-30 is a valid measure for use in Chinese adolescents.

Research on the Association Between Periodontitis and COPD.

Periodontitis is a common chronic bacteria-initiated inflammatory disease that is closely associated with various systemic diseases, including chronic obstructive pulmonary disease (COPD). Periodontitis and COPD share similar risk factors, pathology and microorganisms. Epidemiological and clinical research have shown positive correlation between the two diseases. Individuals with severe periodontitis had a higher risk of developing COPD. Moreover, the relative risk of COPD in severe periodontitis was much higher compared to people without periodontal disease and patients with mild to moderate periodontitis. COPD patients with periodontitis had a higher frequency of COPD exacerbation and periodontal treatment demonstrated some control of COPD. However, the nature of periodontitis affecting COPD still needs further exploration. Periodontitis caused microbial and immune imbalances of the lung through several aspects: (I) under periodontitis status, periodontal pathogens directly caused the lung inflammatory reaction after inhalation and colonization on the lung, (II) periodontitis status promoted the oral colonization of pneumonia-associated pathogens, (III) periodontitis status affected the respiratory epithelium structure and (IV) periodontitis status caused imbalances in neutrophils, macrophages and inflammatory cytokines. In this review, we conclude the association between periodontitis and COPD through several aspects and further discuss the potential mechanism by which periodontitis affects COPD.

Periodontitis contributes to COPD progression via affecting ferroptosis.

BACKGROUND: Periodontitis has emerged as a potential risk factor for chronic obstructive pulmonary disease (COPD). However, the precise mechanism through which periodontitis influences the progression of COPD requires further investigation. Ferroptosis is one of the crucial pathogenesis of COPD and recent researches suggested that periodontitis was associated with ferroptosis. Nonetheless, the relationship among periodontitis, COPD and ferroptosis remains unclear. This study aimed to elucidate whether periodontitis contributes to COPD exacerbation and to assess the potential impact of ferroptosis on periodontitis affecting COPD. METHODS: The severity of COPD was assessed using Hematoxylin and eosin (H&E) staining and lung function tests. Iron assays, malondialdehyde (MDA) measurement and RT-qPCR were used to investigate the potential involvement of ferroptosis in the impact of periodontitis on COPD. Co-cultures of periodontitis associated pathogen Phophyromonas gingivalis (P. gingivalis) and lung tissue cells were used to evaluate the effect of P. gingivalis on inducing the ferroptosis of lung tissue via RT-qPCR analysis. Clinical Bronchoalveolar Lavage Fluid (BALF) samples from COPD patients were collected to further validate the role of ferroptosis in periodontal pathogen-associated COPD. RESULTS: Periodontitis aggravated the COPD progression and the promotion was prolonged over time. For the first time, we demonstrated that periodontitis promoted the ferroptosis-associated iron accumulation, MDA contents and gene expressions in the COPD lung with a time-dependent manner. Moreover, periodontitis-associated pathogen P. gingivalis could promote the ferroptosis-associated gene expression in single lung tissue cell suspensions. Clinical BALF sample detection further indicated that ferroptosis played essential roles in the periodontal pathogen-associated COPD. CONCLUSION: Periodontitis could contribute to the exacerbation of COPD through up-regulating the ferroptosis in the lung tissue.

HSPA5, as a ferroptosis regulator, may serve as a potential therapeutic for head and neck squamous cell carcinoma.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is a ferroptosis sensitive tumor type with high mortality rate. However, it remains largely unknown whether ferroptosis influences the tumor cell in HNSCC. MATERIALS AND METHODS: To investigate how ferroptosis regulators were differentially expressed between normal and tumor tissue, data related to HNSCC was downloaded from The Cancer Genome Atlas. The expression levels of key factors in HNSCC and the relationship between key factors and ferroptosis in HNSCC were conducted in vitro, and then analyzed to correlate with the differences in prognosis and survival. This was then combined with TNM staging data, and the migration effects of key factors in HNSCC were verified by scratch test and transwell test. RESULTS: In this study, gene expression analysis and correlation studies between genes showed that HSPA5 was a potentially key associated ferroptosis regulator in HNSCC. Bioinformatics analysis showed that high expression of HSPA5 in HNSCC was positively correlated with poor prognosis and distal metastasis of HNSCC. In vitro immunohistochemistry and western blot tests confirmed that HSPA5 was highly expressed in HNSCC tissues and cell lines. In vitro inhibition of HSPA5 reduced the viability of HNSCC cells and increased ferroptosis. The results of scratch, transwell, and immunofluorescence tests showed that HSPA5 was related to the migration of HNSCC. In addition, a pan-cancer analysis showed that HSPA5 was also overexpressed in many types of cancer with poor prognoses. CONCLUSION: In total, our study demonstrates the critical role of ferroptosis regulators in HNSCC and that HSPA5, as a ferroptosis regulator, can be regarded as a key molecular target for designing new therapeutic regimens to control HNSCC metastasis and progression.

The remineralization effect of GERM CLEAN on early human enamel caries lesions in vitro.

This study aimed to evaluate the remineralization effect of GERM CLEAN, a novel antibacterial peptide, on early enamel caries. Thirty human enamel blocks from thirty teeth were randomly divided into three groups: double distilled water (DDW group), GERM CLEAN (GC group), and 1000 ppm fluoride (NaF group). Specimens were demineralized for 3 days (pH 4.6) followed by pH cycling twice daily for 14 days. For a pH cycle, specimens received corresponding treatments for 5 min, then were immersed in demineralizing solution for 1 h, received corresponding treatments again, and finally were immersed in remineralizing solution (pH 7.0) for approximately 11 h. Specimens were washed with DDW after each treatment. Microindentation tests, atomic force microscopy (AFM), and transverse micro-radiography (TMR) were conducted to analyze enamel blocks. GC demonstrated a lower percentage of surface microhardness recovery (SMHR%) (p < 0.0001), rougher surfaces (p < 0.0001), deeper lesion depth (p = 0.001), and more mineral loss (p = 0.001) than NaF, but showed higher SMHR% (p < 0.0001), smoother surfaces (p < 0.0001), shallower lesion depth (p = 0.049), and less mineral loss (p = 0.001) than DDW. As a result, GERM CLEAN has the potential to promote the remineralization of demineralized enamel.

Window filtering algorithm for a low repetition rate pulsed laser coherent combination system.

The multi-dithering method has been well verified in the phase-locking of polarization coherent combination experiments. However, it is difficult to apply to low repetition rate pulsed laser coherent combination, since there exists an overlap in the frequency domain between the pulse laser and the large amplitude-phase noise resulting in traditional filters being unable to effectively separate the phase noise. Aiming to solve the problem, we propose, to the best of our knowledge, a novel method of pulse noise detection, identification, and filtering based on the autocorrelation characteristics between noise signals. The self-designed adaptive window filtering algorithm can effectively filter the pulse signal doped in the phase noise around 0.1 ms. After the pulses are filtered out, the remaining phase noise signal is used as the input signal of the multi-dithering method for phase locking; the phase difference of two pulsed beams (10 kHz) is successfully compensated to zero; and the coherent combination of the closed-loop phase lock is realized. Simultaneously, the phase correction periods are short, the phase lock effect is stable, and the intensity of the final combined pulses rises to the ideal value (0.9Imax). In addition, the adaptive window filtering algorithm we proposed can be applied to the coherent combined system of large array fiber lasers and further lay the foundation for fiber phased array lidar.

An in vitro evaluation of antimicrobial activity of a fast-setting endodontic material.

The aim of this study is to evaluate the antimicrobial activity of the fast-setting iRoot Fast Set Root Repair Material (iRoot FS), Mineral trioxide aggregate (MTA) and Biodentine. The materials were freshly mixed or set for 1 and 7 days to conduct the agar diffusion test, direct contact test and carry-over effect test against E. faecalis and P. gingivalis, and the pH values were also measured. The data were analyzed by an analysis of variance and one-way ANOVA or Dunnett's T3 test, and the Tukey's post hoc test for multiple comparisons (α = 0.05). In the direct contact test, all three materials showed good antibacterial activity after setting for 20 min. The antibacterial properties of the three materials decreased with the increase of setting time (p < 0.05). The suspension of all the three materials showed high pH values (11-12) and no significant difference was observed (p > 0.05). With the extension of setting time, the pH of iRoot FS and Biodentine slightly decreased (p < 0.05). Fresh iRoot FS, Biodentine, and MTA killed E. faecalis and P. gingivalis effectively, but their antimicrobial effect decreased after 24 h, and distinctly decreased after 7 days after mixing. iRoot FS, Biodentine, and MTA showed a tendency of alkalinity during this 7-day experiment.

Human periodontitis-associated salivary microbiome affects the immune response of diabetic mice.

Background: The bidirectional association between periodontitis and diabetes mellitus has been well accepted; however, pathways connecting them remain unclear. Some oral bacteria are able to induce immunologic changes favoring insulin resistance individually. However, it is unclear if and how the systemic immune system responds to a disturbed oral microbial community in diabetic sufferers. Aim: This study aimed to investigate the impact of the human periodontitis-associated salivary microbiome on the splenic immune responses of diabetic mice. Methods: An in vivo diabetic animal model was established by feeding high fat food. After microbial depletion with quadruple antibiotic treatment, human saliva from healthy and periodontitis volunteers was transplanted into the mouth of these diabetic mice (N = 3), respectively. Results: Osteoclasts and expression levels of TNF-α and IL-1ß were significantly increased in periodontal tissues of mice receiving periodontitis patients donated microbiome compared to these transplanted with healthy subjects donated microbiome. The proportion of monocyte (an innate immunocyte) decreased in mice receiving periodontitis patients donated microbiome. However, the abundance of an adaptive immunocyte Th17 was up-regulated. The IL17 production of ILC3 cells in human periodontitis-associated salivary microbiome recipient mice was significantly impaired. Conclusions: A disturbed oral microbiome imposes a stress on the splenic immune responses of diabetic mice.

Alteration of intestinal microecology by oral antibiotics promotes oral squamous cell carcinoma development.

Oral antibiotics can influence cancers and immunotherapy by interfering with the intestinal microbiota. However, the association between oral antibiotics and oral squamous cell carcinoma (OSCC) as well as the mechanisms underlying the effects of oral antibiotics on OSCC remain unclear. Here, we found that oral antibiotics cocktail (4Abx) promoted the tumor development and shifted the microbiota, decreasing the abundance of probiotic bacteria, and altered microbial metabolism in the gut of OSCC mice, increasing tyrosine and decreasing glutamate levels. In vitro experiments showed that tyrosine upregulated the PD-1 expression in T cells, SCC7 cell proliferation, and necroptosis expression. IL-10 expression level in CD11c+ cells was reduced by glutamate. Furthermore, the expression of the necroptosis-related proteins, including receptor-interacting protein kinase 1 (RIPK1), RIPK3, and mixed lineage kinase domain-like (MLKL), was higher in the OSCC mice treated with 4Abx. Supplementation with glutamate or healthy mouse feces by gavage alleviated the tumor-promoting effect of 4Abx with restored balance of microbial metabolism. Overall, we identified the detrimental role of oral antibiotics in promoting OSCC development through altered intestinal microbiota, microbial metabolism, and immune dysbiosis, implying the need for antibiotic stewardship in OSCC treatment.

The Arginine Biosynthesis Pathway of Candida albicans Regulates Its Cross-Kingdom Interaction with Actinomyces viscosus to Promote Root Caries.

The cross-kingdom interactions between Candida albicans and Actinomyces viscosus play critical roles in root caries. However, the key pathway by which C. albicans regulates its interactions with A. viscosus is unclear. Here, we first employed 39 volunteers with root caries and 37 caries-free volunteers, and found that the abundances of C. albicans and A. viscosus were significantly increased in the individuals with root caries and showed a strong positive correlation. Their dual-species combination synergistically promoted biofilm formation and root caries in rats. The arginine biosynthesis pathway of C. albicans was significantly upregulated in dual-species biofilms and dental plaques from another 10 root caries volunteers compared with the 10 caries-free volunteers. The exogenous addition of arginine increased the cariogenicity of the dual-species biofilm. The C. albicans ARG4, a key gene from the arginine biosynthesis pathway, null mutant failed to promote dual-species biofilm formation and root caries in rats; however, the addition of arginine restored its synergistic actions with A. viscosus. Our results identified the critical roles of the C. albicans arginine biosynthesis pathway in its cross-kingdom interactions with A. viscosus for the first time and indicated that targeting this pathway was a practical way to treat root caries caused by multiple species. IMPORTANCE Root caries is a critical problem that threatens the oral health of the elderly population. Our results identified the essential roles of the C. albicans arginine biosynthesis pathway in its cross-kingdom interactions with A. viscosus in root caries for the first time and indicated that targeting this pathway was a practical way to treat root caries caused by multiple species.

Anticaries activity of GERM CLEAN in Streptococcus mutans and Candida albicans dual-species biofilm.

OBJECTIVE: To evaluate the antimicrobial effects of a peptide containing novel oral spray GERM CLEAN on dual-species biofilm formed by Streptococcus mutans and Candida albicans and to investigate whether GERM CLEAN inhibits the demineralization procedure of bovine enamel in vitro. METHODS: The antimicrobial effects of GERM CLEAN on dual-species biofilm were analyzed by initial adherence rate calculation, water-insoluble exopolysaccharides quantification, total biomass quantification, and colony-forming units (CFUs) counting. Scanning electron microscopy and confocal laser scanning microscopy were applied to evaluate the impacts of GERM CLEAN on the biofilm structure. Further, the effects of GERM CLEAN on acidogenicity of dual-species were appraised via glycolytic pH drop analysis and hydroxyapatite dissolution measurement. The percentage of Surface Microhardness Reduction (%SMHR) evaluation, Atomic Force Micrograph (AFM) examination, and Transverse Microradiography (TMR) analysis after pH cycling were used to determine whether GERM CLEAN inhibited the demineralization of bovine enamel. RESULTS: GERM CLEAN decreased the adherence rate, water-insoluble EPS production, biofilm formation, and acidogenicity of the dual-species. Moreover, GERM CLEAN significantly inhibited the demineralization status of bovine enamels. CONCLUSION: This peptide containing novel oral spray GERM CLEAN has antimicrobial potential toward the dual-species. GERM CLEAN can also impede the demineralization procedure of enamel.

Antimicrobial Effect of a Peptide Containing Novel Oral Spray on Streptococcus mutans.

OBJECTIVE: To investigate the antibacterial effect of a novel antimicrobial peptide containing oral spray GERM CLEAN on Streptococcus mutans (S. mutans) in vitro and further explore the related mechanisms at phenotypic and transcriptional levels. METHODS: The disk diffusion method was used to preliminarily appraise the antimicrobial effect of GERM CLEAN. The minimal inhibitory concentration (MIC) of GREM CLEAN towards S. mutans was determined by the broth dilution method. S. mutans was determined by the broth dilution method. RESULTS: The diameter (10.18 ± 1.744 mm) of inhibition zones formed by GERM CLEAN preliminarily indicated its inhibitory effect on the major cariogenic bacteria S. mutans was determined by the broth dilution method. S. mutans was determined by the broth dilution method. S. mutans was determined by the broth dilution method. S. mutans was determined by the broth dilution method. gtfB, gtfC, gtfD, and ldh were significantly repressed by treating with GERM CLEAN, and this was consistent with our phenotypic results. CONCLUSION: The novel antimicrobial peptide containing oral spray GERM CLEAN has an anti-Streptococcus mutans effect and the inhibitory property may be due to suppression of the virulence factors of S. mutans including adhesive, acidogenicity, EPS, and biofilm formation.Streptococcus mutans effect and the inhibitory property may be due to suppression of the virulence factors of S. mutans including adhesive, acidogenicity, EPS, and biofilm formation.S. mutans was determined by the broth dilution method.

Sortase A-mediated modification of the Streptococcus mutans transcriptome and virulence traits.

Sortase A contributes to adhesion and biofilm formation of Streptococcus mutans by anchoring surface proteins like P1 onto the cell wall, and few other functional characterization has been annotated to this protein and its coding gene srtA. In this study we investigated that whether srtA deletion would affect S. mutans virulence determinants in addition to adhesion and further explored whether these effects were caused due to changes in S. mutans genomic transcription. We used acid-killing assays, glycolytic rate assessments, and exopolysaccharide (EPS) formation tests to detect whether srtA deletion influenced S. mutans acid tolerance/production and glucan formation. Comparisons between RNA-sequencing data from both the exponential and stationary phases of UA159 and the srtA-deleted strain were made to determine the impact of srtA knockout on S. mutans genomic transcription. Results of our assays indicated that S. mutans aciduricity was enhanced in the srtA deleted strain when bacterial cells were directly subjected to pH 2.8, but the enhancement was repressed when the acid tolerance response was induced in advance. The srtA mutation strain exhibited reduced EPS formation in mature biofilms. SrtA deletion led to pleiotropic changes in the S. mutans transcriptome with a growth phase-dependent pattern. The affected genes mainly included those involved in aciduricity, carbohydrate transport, and EPS formation. It was concluded that S. mutans srtA exhibited multiple effects on the virulence traits of this pathogen, including acid tolerance and glucan formation, and that these alterations could be partially due to transcriptional changes upon loss of srtA.

Research progress in laser treatment of dentin hypersensitivity / 口腔疾病防治

@#Dentin hypersensitivity is mainly associated with abrasion, wear, acid etching, cracking, wedge-shaped defects, enamel hypoplasia, caries, a lack of neck enamel, and cementum coverage accompanied by gingival recession, resulting in direct exposure of dentin tubules to the oral environment. Dentin hypersensitivity is mainly treated by sealing the exposed dentin tubules and reducing the excitability of the pulpal nerves. Laser therapy, as a safe, fast and convenient treatment measure, has achieved good results both alone and in combination with other medicines and has attracted increasing attention. This paper reviews the research progress in the treatment of dentin hypersensitivity using several common laser therapy mechanisms with parameter selections in various scopes of application. A review of the literature shows that the Nd:YAG laser has a strong penetrating power and a large thermal effect and can denature and coagulate proteins in dentin tubules in a very short time; while the CO2 laser causes little damage to the dental pulp and has an obvious immediate curative effect by high absorption of water molecules in hydroxyapatite and the dentin surface is melted and recrystallized; the Er:YAG laser has high water absorption and a large thermal effect and can block the tubules by evaporating the fluid in the tubules and depositing salts; Er, Cr:YSGG laser energy can be fully absorbed by hydroxyapatite and water and result in desensitization by cutting hard tissues and melting periodontal dentin of the tube simultaneously; The Ga-Al-As semiconductor laser and He:Ne laser are low energy. The laser dosage mainly changes the permeability of nerve fibers to potassium and sodium ions and depolarizes the nerve fibers, producing an analgesic effect. The combination of a laser with a desensitizer can improve the clinical efficacy of treating dentin hypersensitivity.

Overexpression of Nmnat improves the adaption of health span in aging Drosophila.

Nmnat is a stress response protein which has been involved in a variety of biological processes. However, the effects of Nmnat on aging have not yet been investigated. The present study revealed the effects of Nmnat on aging of Drosophila and uncovered its underlying mechanism. Therefore, the overexpression of Nmnat was established by arm/Gal4 system in Drosophila with an aim to determine the functions of Nmnat during aging process. In this study, our results showed Nmnat was a positive factor on lifespan and movement capacity, which was consistent on d-galactose induced aging acceleration. Further investigation showed that oxidative stress biomarkers, longevity gene, mitochondria related genes and ATP levels were significantly improved in the Nmnat overexpression Drosophila, which suggested that the underlying mechanism of Nmnat on aging process and movement capacity was partly due to its anti-oxidative stress and mitochondrial-protection function. In addition, on H2O2 challenge tests, Nmnat overexpression was sufficient to increase the survival time and movement capacity of Drosophila, which was probably due to its protection against oxidative stress. On rotenone induced mitochondrial dysfunction, Nmnat overexpression also showed better health span and movement capacity than the control group. Based on these data, Nmnat may be a new molecular target to improve health span by enhancing stress resistance and locomotor activity in aging process.