Objective:To investigate the effects of berberine on the proliferation,apoptosis,inflammation,and osteogenic differen-tiation of human periodontal ligament stem cells(hPDLSCs)induced by lipopolysaccharide through regulating the NF-κB/NLRP3 signaling pathway.Methods:HPDLSCs with good growth status were separated into a blank group(without any treatment),a lipopolysaccharide group(1 μg/mL lipopolysaccharide,A group),a low concentration berberine(berberine-L)group(1 μg/mL lipopolysaccharide+25 μmol/L berberine,B group),a medium concentration berberine(berberine-M)group(1 μg/mL lipopolysac-charide+50 μmol/L berberine,C group),a high concentration berberine(berberine-H)group(1 μg/mL+100 μmol/L berberine,D group),and a berberine-H+Prostratin group(1 μg/mL lipopolysaccharide+100 μmol/L berberine+1 μmol/L Prostratin,E group).Subsequently,the cell proliferation,levels of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)in the supernatant,cell apoptosis,osteogenic differentiation ability,expression of osteogenic differentiation related genes[RUNX family transcription fac-tor 2(Runx2),osteopontin(OPN),osteocalcin(OCN)],and expression levels of NF-κB and NLRP3 mRNA were detected.Results:The number of mineralized nodules,cell proliferation rate,ALP content,OPN,OCN,and Runx2 protein expression in the lipopolysac-charide group were greatly reduced compared to the blank group,however,the levels of IL-1 β,IL-6,and TNF-α,apoptosis rate,NF-κB,and NLRP3 mRNA expression in the supernatant were greatly increased(P＜0.05).After treating cells with different doses of berberine,the number of mineralized nodules,proliferation rate,ALP content,OPN,OCN,and Runx2 protein expression were greatly increased,the levels of IL-1β,IL-6,and TNF-α,apoptosis rate,NF-κB,and NLRP3 mRNA expression in the supernatant were greatly reduced,and the berberine-H group was the most great(P＜0.05).After treatment with berberine-H combined with Prostratin,the number of mineralized nodules,proliferation rate,ALP content,OPN,OCN and Runx2 protein expression were greatly reduced compared to single berberine-H treatment,the levels of IL-1β,IL-6 and TNF-α,apoptosis rate,NF-κB,and NLRP3 mRNA expression in the supernatant were greatly increased(P＜0.05).Conclusion:Berberine enhances lipopolysaccharide induced proliferation of hPDLSCs,inhibits apoptosis and inflammation,and promotes osteogenic differentiation by inhibiting the NF-κB/NLRP3 signaling pathway.

In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

Objective:To explore the effects of parathyroid hormone(PTH)on the osteogenic differentiation of osteoblasts by reg-ulating macrophage polarization in inflammatory microenvironment.Methods:Macrophages were pretreated with lipopolysaccharide(LPS)for 2 h to establish an inflammatory microenvironment model,and then treated with PTH for 24 h.Macrophages and osteo-blasts were co-cultured in Transwell cells.Alkaline phosphatase staining,alizarin red staining,RT-qPCR and Western blot were applied to detect osteogenic differentiation.The expression of SOCS1/JAK2/STAT3 protein in macrophages was detected by West-ern blot.The change of STAT3 expression was detected after adding AG490.The expression of miR-155-5p,SOCS1,IL-1β,IL-6 and i-NOS was detected by ELISA and RT-qPCR.Results:LPS induced M1-type polarization of macrophages and inhibited the osteogenic differentiation of osteoblasts.PTH inhibited the polarization of M1-type macrophages and promoted the osteogenic differ-entiation of osteoblasts in inflammatory microenvironment(P＜0.05).PTH down-regulated the expression of miR-155-5p,IL-1β,IL-6,i-NOS,p-JAK2/JAK2 and p-STAT3/STAT3 in macrophages under inflammatory microenvironment(P＜0.05),and up-reg-ulated SOCS1(P＜0.05).AG490 further inhibited p-STAT3/STAT3 expression.Conclusion:PTH inhibits the polarization of M1-type macrophages and promotes osteogenic differentiation of osteoblasts by down-regulating miR-155-5p and then targeting SOCS1/JAK2/STAT3 signaling pathway in inflammatory microenvironment.

Objective:To investigate the function and mechanism of exosome Linc00665 in modulating CD8+T cell immunoreactivity to promote radiotherapy resistance in OSCC.Methods:HOEC,SCC9 and SCC9-RR exosomes were extracted and identified,and the expression of Linc00665 was detected by qRT-PCR in cell lines and exosomes.The expression of TNF-α,IFN-γ,perforin and granzyme B in each treatment group was detected by ELISA(PBS,SCC9 exo,SCC9-RR exo).The killing ability of CD8+T cells against SCC9 cells in each treatment group was detected by CCK-8 assay.The targets of Linc00665 were further bioinformatically ana-lyzed and verified by qRT-PCR and Western blot.The expression of Linc00665,miR-28-5p and PD-1 in CD8+T cells was exogenous-ly regulated,the expression of immunoreactive factors in the supernatants of each treatment group was detected by ELISA(NC,sh-Linc00665,miR-28-5p inhibitor,sh-PD-1),and the killing ability of cells in each group was detected by CCK-8 method.Results:The concentrations of TNF-α,IFN-γ,perforin and granzyme B in the supernatants of cell culture in the SCC9-RR exo/CD8+T group were significantly decreased compared with those in the PBS/CD8+T group and the SCC9 exo/CD8+T group(P＜0.05),and the kill-ing ability of the cells in the SCC9-RR exo group was significantly decreased compared with those in the PBS group and the SCC9 exo group(P＜0.05),suggesting that SCC9-RR exo could inhibit the tumor killing ability of CD8+T cells.qRT-PCR results suggested that Linc00665 was highly expressed in the SCC9-RR cell line as well as exosome(P＜0.05).It was further verified by bioinformat-ics analysis that Linc00665 could regulate PD-1 expression via miR-28-5p,thereby modulating CD8+T cell immunoreactivity to pro-mote OSCC radiotherapy resistance.Conclusion:Exosome Linc00665 regulates CD8+T cell immunoreactivity through miR-28-5p/PD-1 axis to promote OSCC radiotherapy resistance.

Objective:To explore the effects of LncRNA DLEU2 on the migration and proliferation of oral squamous cell carcino-ma(OSCC)cells by regulating the miR-186-5p/insulin-like growth factor 2 mRNA binding protein 3(IGF2BP3)axis.Methods:QRT-PCR and Western blot were applied to detect the mRNA and protein levels of LncRNA DLEU2,miR-186-5p,and IGF2BP3 in OSCC cell line SCC-25 and human normal oral keratinocyte line NOK,respectively.SCC-25 cells were transfected with si-DLEU2,miR-186-5p inhibitor and negative control respectively.The relationship between LncRNA DLEU2,miR-186-5p and IGF2BP3 was verified by double luciferase reporter gene assay.qRT-PCR was applied to detect the mRNA expression of LncRNA DLEU2 and miR-186-5p in SCC-25 cells.CCK-8 test was applied to detect the proliferation of SCC-25 cells.Flow cytometry was applied to detect the apoptosis rate of SCC-25 cells.Transwell assay was used to detect cell invasion and migration.Western blot was used to detect EMT-associated and IGF2BP3 protein levels.Results:Silencing LncRNA DLEU2 decreased the proliferative ac-tivity,migration and invasion of SCC-25 cells and decreased the protein levels of N-cadherin and Vimentin,increased the apopto-sis rate,miR-186-5p expression and E-cadherin protein levels.Down-regulation of miR-186-5p weakened the inhibitory effect of silenced LncRNA DLEU2 on the malignant phenotype of SCC-25 cells.LncRNA DLEU2 negatively regulated the miR-186-5p/IGF2BP3 axis.Conclusion:Silencing LncRNA DLEU2 may down-regulate the expression of IGF2BP3 by up-regulating miR-186-5p,inhibites the proliferation,migration,and invasion of SCC-25 cells,and promotes the apoptosis of the cells.

Objective:To study the efficiency and biosafety of Electrochemical advanced oxidation processes(EAOP)in dental bleaching,and conduct preliminary application.Methods:Indigo carmine and coffee were used as the indicator to assess the effi-cacy of EAOP.High resistance meter was used to measure the resistance of the tooth to verify the safety of the working voltage.Twenty wisdom teeth after tooth extraction were collected,dyed and bleached in vitro to verify the bleaching efficiency.Subsequent-ly,the bleached teeth were examined by scanning electron microscopy,hardness testing,and bacterial adhesion experiments to as-sess surface damage.To determine its cytotoxicity,cells were co-cultured with electrolyte.Initial samples of bleaching tray was prepared,and its durability were verified.Results:The EAOP could bleach indigo carmine within 10 min and coffee within 90 min at an operating voltage of 8 V.The resistance at the groove of the tooth socket was(3.4±1.2)MΩ,and the theoretical calculated current was less than 3 μA.The efficiency of EAOP tooth bleaching was slightly lower than that of traditional office bleaching and higher than that of home bleaching.Compared with the traditional bleaching method,scanning electron microscopy showed that EAOP had less demineralization effect on tooth surface.The tooth hardness before and after bleaching had no statistical difference(P=0.912).The bacterial adhesion test after tooth bleaching showed that EAOP method could reduce about 60％bacterial adhe-sion(P＜0.001).The cytotoxicity test showed that EAOP electrolyte had no obvious toxic effect.The durability test shows that the bleached denture still has good bleaching effect after 20 h of use.Conclusion:Compared with the traditional bleaching method,EAOP bleaching had excellent tooth bleaching effect,little effect on tooth damage,high safety,and the related bleaching devices had good durability.

Objective:To investigate the possibility of 1,8-cineole in the prevention of Streptococcus mutans(S.mutans)induced dental caries in rats.Methods:40 specific pathogen-free rats were randomly divided into 5 groups(n=8):distilled water group(negative control group),chlorhexidine group(positive control group),and eucalyptin low(MIC),medium(2MIC)and high(4MIC)dose groups.The plates were streaked with saliva samples,and calculated the number of S.mutans colonies and the ratio of the total number of colonies.Keyes' method was used to evaluate the caries damage of the jaws specimens.Results:At the end of the experiment,the proportion(％)of S.mutans in the oral cavity of the drug groups decreased(P＜0.000 1),but there was no significant difference between the drug group and the positive control group(P＜0.05).At the end of the experiment,compared with the negative control group,the scores of each grade in the drug group decreased(P＜0.05),but there was no significant differ-ence between positive control group and each 1,8-cineole group(P＞0.05),and among the 3 1,8-cineole dose groups(P＞0.05).Conclusion:1,8-cineole at the MIC can inhibit the growth of S.mutans in the oral cavity of rats with dental caries and reduce the frequency and severity of dental caries.

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective:To explore the mechanism of long non-coding RNA(lncRNA)HOXA transcript at the distal tip(HOT-TIP)in promoting the proliferation,migration and invasion of oral cancer cells via the mitogen activated protein kinase/extracellu-lar regulated protein kinase(MAPK/ERK)pathway.Methods:Human oral cancer KB cells were cultured in vitro and divided into control(without any treatment),pcDNA(transfected with pcDNA-NC,A group),pcDNA-HOTTIP(transfected with pcDNA-HOT-TIP,B group),MAPK/ERK pathway inhibitor PD98059(PD98059 group,50 μmol/L PD98059,C group)and pcDNA-HOTTIP+PD98059 groups(transfected with pcDNA-HOTTIP+50 μmol/L PD98059,D group).qRT-PCR was applied to detect HOTTIP mRNA expression,scratch test was applied to detect KB cell migration,Transwell assay was applied to detect cell invasion,flow cytometry was applied to detect cell apoptosis,CCK-8 was used to detect cell proliferation,Western blot was applied to detect the MAPK/ERK signaling pathway and epithelial mesenchymal transition(EMT)related protein expression levels in KB cells.Results:Compared with the control group,the KB cell scratch healing rate,cell invasion number,A450 value,p-MEK1/2,p-ERK1/2,and N-cadherin expression in pcDNA-HOTTIP group increased,the apoptosis rate and E-cadherin expression reduced(P＜0.05);scratch healing rate,cell invasion number,A450 value,p-MEK1/2,p-ERK1/2,and N-cadherin expression in PD98059 group reduced,the apoptosis rate and E-cadherin expression increased(P＜0.05).PD98059 weakened the role of HOT-TIP in promoting proliferation,migration and invasion of KB cells.Conclusion:LncRNA HOTTIP promotes proliferation,migra-tion,and invasion of KB cells by activating the MAPK/ERK pathway.

Objective:To investigate the changes of upper airway and adenoids in children with hypertrophic adenoid skeletal classⅠ and class Ⅲ anterior crossbite after orthodontic treatment.Methods:From January 2021 to January 2022,155 children with skeletal class Ⅲ anterior crossbite who were treated with anterior traction in Stomatological Hospital,Shanxi Medical University were selected.They were divided into class Ⅲ normal group(72 cases)and class Ⅲ hypertrophic group(83 cases)according to whether the adenoids were hypertrophic.A total of 122 children with class Ⅰ anterior counterocclusion were treated with"2×4"treatment,which were di-vided into Class Ⅰ normal group(60 cases)and class Ⅰ hypertrophy group(62 cases).The changes of upper airway and adenoid were compared between the two groups.Results:(1)After intervention,only the width of nasopharynx cavity increased(P＜0.05)in Class Ⅰ normal group and Class Ⅰ hypertrophic group,and there was no significant difference in other indicators(P＞0.05).(2)After intervention,the adenoid thickness and A/N ratio of children with skeletal class Ⅲ anterior crossbite decreased,and the width of na-sopharynx cavity,nasopharynx airway space,soft palate upper and rear airway space,uvula tip rear airway space,the minimum air-way space between soft palate and adenoid,and mandibular plane angle increased(P＜0.05).The A/N ratio of class Ⅲ hypertrophic group was lower than that of class Ⅲ normal group(P＜0.05).Conclusion:Orthodontic treatment can reduce the degree of adenoid hypertrophy and expand the airway in children with skeletal class Ⅲ anterior crossbite.Only the width of the nasopharynx cavity can be widened after the correction intervention for children with adenoid hypertrophy and skeletal class Ⅰ anterior crossbite.