To study the hypoxia response gene and microRNA (miRNA) expression profiles in the pathogenesis and progression of oral leukoplakia (OLK).

Affymetrix GeneChip human transcriptome array 2.0 was used to detect the transcriptome of normal mucosa, low-risk OLK, high-risk OLK, and early squamous cell carcinoma (SCC). Gene ontology function analysis was used to screen genes and key miRNAs whose biological role is hypoxia response. Quantitative reverse transcription polymerase ch-ain reaction (qRT-PCR) was used to verify the expression of hypoxia response genes and miRNAs.

A total of 7 different genes of hypoxia response between normal mucosa and low-risk OLK, 10 genes between low-risk and high-risk OLK, and 21 genes between high-risk OLK and SCC were identified. The results of qRT-PCR showed that the expression of hypoxia-inducible factor 1α, chemokine cc-motif ligand 2, and matrix metalloproteinase 3 mRNA and miR-21 in normal mucosa, OLK, and SCC increased in a stepwise manner. The expression difference between OLK and SCC was statistically significant and consistent with the results of transcriptome array.

The hypoxia response gene and related miRNA play roles in the development and progression of OLK.

This study aimed to investigate the effects of microRNA-146a (miR-146a) on the production of cytokines in lymphocytes stimulated by Porphyromonas gingivalis (P.gingivalis) lipopolysaccharide (LPS).

Lymphocytes were harvested from mouse spleen and cultured in vitro. The cells were treated with P. gingivalis LPS, miR-146a mimic, or miR-146a inhibitor. Scramble RNA served as the negative control of mimic and inhibitor. The production of inflammatory cytokines was detected by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay.

Compared with non-LPS-stimulated group, P. gingivalis LPS could increase the levels of interleukin (IL)-1β, IL-6, receptor activator NF-κB ligand (RANKL), and IL-10 (P<0.05) and decrease the mRNA level of osteoprotectin (OPG) (P<0.05). However, it did not significantly change the secretion of OPG. Compared with the negative control group, miR-146a mimic upregulated the levels of IL-10 and OPG (P<0.05), downregulated IL-1β, IL-6, and RANKL (P<0.05). Meanwhile, miR-146a inhibitor had a reverse effect on these cytokines (P<0.05) in P.gingivalis LPS-treated-lymphocytes.

MiR-146a can provide a suitable microenvironment for bone formation by preventing the inflammatory effects of P.gingivalis LPS through the inhibition of IL-1β, IL-6, and RNAKL, thereby enhancing IL-10 and OPG.

A study was conducted to explore the expression pattern and function of ferritin heavy polypeptide gene (fth1b) in zebrafish pharyngeal teeth development and lay the foundation for subsequent research on teeth development and mineralization.

The zebrafish embryos were harvested at 56, 72, 96, and 120 h after ferti-lization. The expression of fth1b in zebrafish pharyngeal teeth development was detected by whole embryo in situ hybri-dization and compared with the known pharyngeal teeth marker dlx2b. The specific knockout of fth1b gene was performed using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing technology. The development of zebrafish pharyngeal teeth was detected in the fth1b^-/- mutant.

The expression pattern of fth1b gene was very similar to that of the known zebrafish pharyngeal teeth marker dlx2b and was specifically expressed in the zebrafish pharyngeal teeth during development. After the specific knockout of the gene fth1b, the earliest gene that can be detect in zebrafish pharyngeal teeth-pitx2 was expressed normally during early development. The dlx2b expression was not significantly different from that of wild type zebrafish, but the mineralization of pharyngeal teeth in the mutant was weaker than that of wild type zebrafish.

The gene fth1b is specifically expressed in zebrafish pharyngeal teeth and acts on their early mineralization.

This study aimed to assess the influence of different types of rapid maxillary expansion on root resorption (RR).

Literature searches were carried out electronically in five English and two Chinese databases. Randomized controlled trials (RCTs), controlled clinical trials (CCTs), cohort studies, and case-control studies were included. The data were extracted by three authors. The risk of bias in the RCTs and nonrandomized studies were assessed in accordance with corresponding scales.

Among the 400 articles identified, seven were included for the final analysis. Three studies were graded as high value of evidence, while two and another two studies were graded as moderate value and low value, respectively. According to the available evidence, the tooth-borne maxillary expansion caused more obvious RR of anchorage teeth than the bone-borne one. In addition, the Haas-type palatal acrylic pads could not effectively reduce the degree of RR. The difference in the design of the retainer between the tooth-borne maxillary expansion (the use of a band or wire framework to connect the anchorage tooth) did not cause the difference in the incidence and degree of RR.

Clinical evidence suggested that bone-borne maxillary expansion may decrease the amount of RR, while the amounts of resorption did not significantly differ between Haas and Hyrax and between different retainer types of Hyrax.

To assess the treatment outcome of patients who completed their orthodontic treatment by using Andrews' six elements and the American Board of Orthodontic (ABO) objective grading system (OGS); to determine whether Andrews' six elements can be used as a new assessment system for patients who completed their orthodontic treatment.

A total of 160 patients who completed their orthodontic treatment were included in the study. The participants were randomly selected from patients who completed their orthodontic procedures in Kunming Medical University Affiliated Stomatological Hospital during the period of 2015 to 2019. The retrospective completed cases were examined in accordance with the Andrews' six elements and ABO measuring scales. Scores were assigned to each tooth in each category. All the measurement items in both evaluation criteria, the composite category score, and the total score were calculated. The passing and potential passing rates of the completed cases were compared with two measuring scales via the Chi-square test.

The passing rate for the evaluation of cases by using the Andrews' six elements measuring scale was 83.8%, and that for the evaluation of cases by using the ABO measuring scale was 86.3%. The differences in achieving the standard between the cases of Andrews' six elements and ABO-OGS via the Chi-square test were statistically insignificant (P>0.05). The mean Andrews' six elements score for the completed cases: the category of the anteroposterior change in position of the incisors contributed the most, whereas the core discrepancy presented the least percentage in total scores. In ABO-OGS, alignment and marginal ridges contributed the most, whereas interproximal contacts exhibited the least percentage in total scores.

The performance of Andrews' six elements was comparable with that of ABO-OGS in assessing the treatment outcome of patients who completed their orthodontic treatment. Andrews' six elements can be used as a new system for assessing the treatment outcome of patients who completed their orthodontic treatment. It demonstrated particular advantage in controlling facial profile, and had just completed material of patients who completed their orthodontic treatment it can measure. The Andrews' six elements measuring scale is convenient to disseminate and use.

To investigate caries status and its impact factors in preschool children in plateau and to provide reference for caries prevention in highlands.

Examination of caries was performed on 1 597 children aged 3-5 years old in 11 kindergartens in Changdu, Xizang, in accordance with the 4th National Oral Health Survey standards and methods. Their parents were surveyed with the questionnaire regarding oral hygiene habit and consciousness about oral health and related factors. All the data were collected and analyzed.

The prevalence of caries among children aged 3-5 years old in Changdu was 52.85%, with dmft index of 2.44. The caries rate and dmft of children aged 3 years old were lower than those of children aged 4 and 5 years old (P<0.05). No significant difference was observed in the caries rate between males and females (P>0.05). Single-factor analysis showed that the frequency of brushing teeth more than twice a day, low frequency of eating sweets, high frequency of drinking butter tea, and regular oral examination can reduce the rate of caries (P<0.05). Multivariate logistic analysis showed that the frequency of eating sweets, drinking butter tea, and oral examination are related impact factors of caries.

The prevalence of ca-ries in children aged 3-5 years old in Changdu increases with aging. Good oral hygiene and eating habits and regular oral examination can reduce the rate of caries.

This study aimed to investigate the association between periodontal indexes and biomarkers in gingival crevicular fluid (GCF) and preterm birth (PTB) in pregnancy, as well as to assess the clinical value of these indexes as predictors of PTB.

A nested case-control study was conducted. A total of 300 systematically healthy pregnant women were selected within 36 weeks of gestation and grouped according to the enrolled weeks. Periodontal indexes, including probing depth (PD), bleeding index (BI), gingival index (GI), and five biomarkers in GCF, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), prostaglandin E2 (PGE2), and 8-hydroxy-2-deoxyguanosine (8-OHdG) were measured at the enrolled date. The detailed birth outcome was recorded.

Only women at 24-28 weeks of gestation per PTB case (four full-term births) were selected as controls subjects, PTB displayed significantly greater GI, BI, and 8-OHdG (P<0.05). Logistic regression analysis revealed that BI and 8-OHdG were the dependent risk factors of PTB (OR=5.90, P=0.034; OR=1.18, P=0.045, respectively). The areas under the receiver operating characteristic curve (ROC) of BI and 8-OHdG were 0.80 and 0.69, and that of the combined detection was 0.82, which was larger than the individual detection, although the differences were not significant (P>0.05).

Increased BI and 8-OHdG at 24-28 weeks of gestation are risk factors for PTB. Their combined detection may have some value in the prediction of PTB, but further studies with a larger sample size are needed to explore it and thus provide experiment evidence for establishing an early warning system for PTB in pregnant women with periodontal disease.

This study aimed to explore the effects of silencing isoprenylcysteine carboxyl methyltransfe-rase (Icmt) through small interfering RNA (siRNA) interference on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC).

Three siRNA were designed and constructed for the Icmt gene sequence and were then transfected into TSCC cells CAL-27 and SCC-4 to silence Icmt expression. The tested cells were divided as follows: RNA interference groups Icmt-siRNA-1, Icmt-siRNA-2, and Icmt-siRNA-3, negative control group, and blank control group. The transfection efficiency of siRNA was detected by the fluorescent group Cy3-labeled siRNA, and the expression of Icmt mRNA was screened by quantitive real-time polymerase chain reaction (qRT-PCR) selected the experimental group for subsequent experiments. The expression of Icmt, RhoA, Cyclin D1, p21, extracellular regulated protein kinases (ERK), and phospho-extracellular regulated protein kinases (p-ERK) were analyzed by Western blot. The proliferation abilities of TSCC cells were determined by cell counting kit-8 assay. The change in apoptosis was detected by AnnexinV-APC/propidium staining (PI) assay. Cell-cycle analysis was conducted by flow cytometry.

The expression of Icmt mRNA and protein in TSCC cells significantly decreased after Icmt-siRNA transfection (P<0.05). No significant difference in RhoA mRNA and protein expression was detected (P>0.05), but the expression of RhoA membrane protein decreased compared with the negative control group and blank control groups (P<0.05). Cyclin D1 expression decreased, whereas p21 expression significantly increased and the relative expression of ERK protein in the experimental group did not significantly different that in the control group (P>0.05). However, the phosphorylation level of ERK was significantly reduced (P<0.05). The cell cycles of TSCC CAL-27 and SCC-4 were altered in G1/S, cell proliferation activity was inhibited, and apoptosis was induced (P<0.05).

Silencing Icmt can effectively downregulate its expression in TSCC cells, reduce the RhoA membrane targeting localization and cell proliferation, and induce apoptosis. Thus, Icmt may be a potential gene therapy target for TSCC.

This study aimed to explore the effect of sex determining region Y-box 9 (SOX9) on the microtubule formation and epithelial-mesenchymal transition (EMT) of human oral squamous cell carcinoma (OSCC) CAL27 and the underlying mechanism.

SOX9-shRNA1 and SOX9-shRNA2 were designed and synthesized and then transfected into CAL27 cells. The expression of SOX9 was detected by quantitative real-time polymerase chain reaction. Microtubule formation assay was used to detect the change in the number of microtubule nodules after interfering with SOX9. Immunofluorescence was used to detect the Vimentin content. Western blot was used to detect the protein expression of EMT marker molecules and Wnt/β-catenin pathway-related proteins, such as E-cadherin, N-cadherin, Fibronectin, Wnt, β-catenin, T-cell factor-4 (TCF-4).

The expression level of SOX9 significantly decreased after transfection with SOX9-shRNA1 and SOX9-shRNA2 in CAL27 cells (F=578.000, P=0.000; F=96.850, P=0.000). Interference with SOX9 inhibited the EMT of OSCC. After interference with SOX9, the number of tubules and Vimentin positive cells decreased significantly (F=169.700, P=0.000). The expression level of E-cadherin significantly increased (F=181.400, P=0.000). The expression levels of N-cadherin, Fibronectin, Wnt, β-catenin, and TCF-4 proteins significantly decreased (N-cadherin: F=101.400, P=0.000; Fibronectin: F=122.300, P=0.000; Wnt: F=70.290, P=0.000; β-catenin: F=81.740, P=0.000; TCF-4: F=37.020, P=0.000).

Interference with SOX9 decreased Vimentin content and inhibited the microtubule formation and protein expression of EMT marker molecules, as well as the expression of proteins related to the Wnt/β-catenin pathway. Thus, SOX9 can induce microtubule formation and EMT in CAL27, which was related to the inhibition of the Wnt/β-catenin pathway activation.

A study was conducted to investigate the molecular mechanism of chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) influencing the invasion and metastasis of tongue squamous cell carcinoma and to provide a new target for clinical inhibition of invasion and metastasis of tongue squamous cell carcinoma.

Ualcan website was used to analyze the expression of CHD1L in normal epithelial tissue and primary head and neck squamous cell carcinoma and to analyze the effect of lymph node metastasis on the expression of CHD1L in tissues with head and neck squamous cell carcinoma. The relationship between CHD1L expression and the survival rate of patients with head and neck squamous cell carcinoma was tested by the GEPIA website. Western blot was used to quantify the levels of CHD1L protein in human tongue squamous cell carcinoma CAL27 and immortalized human skin keratinocyte cell HaCaT. After knocking down CAL27 in human tongue squamous cell carcinoma cells with an RNA interference plasmid, the cells were designated as SiCHD1L/CAL27 and Scr/CAL27. Western blot was utilized to detect the expression of CHD1L in each group of cells. The change in CAL27 cell proliferation ability was tested by EdU proliferation test after CHD1L knockdown. The change of cell migration ability of each group cells was tested through the wound healing assay. Western blot was used to detect epithelial-mesenchymal transition (EMT) marker E-cadherin and Vimentin protein expression levels.

Ualcan database showed that the expression of CHD1L in primary head and neck squamous cell carcinoma tissues was higher than in normal epithelial tissues and in head and neck squamous cell carcinoma tissues with lymph node metastasis. GEPIA website analysis showed that the overall survival rate of patients with head and neck squamous cell carcinoma with high expression of CHD1L was significantly lower than that of patients with low expression. Western blot results showed that CHD1L expression in human tongue squamous carcinoma cells CAL27 was higher than that of human normal skin cells HaCaT. CHD1L expression in SiCHD1L/CAL27 cells was much lower than that in Scr/CAL27 cells. Results of EdU proliferation experiments showed the significant reduction in the cell proliferation ability of the SiCHD1L/CAL27 cells. Results of the wound healing experiments showed the reduction in the migration capacity of the SiCHD1L/CAL27 cells. The expression of E-cadherin increased, whereas that of Vimentin decreased, in SiCHD1L/CAL27 cells.

CHD1L promoted the EMT, proliferation, migration, and invasion ability of tongue squamous cell carcinoma cells.