SUPPRESSOR OF FRIGIDA 4 cooperates with the histone methylation reader EBS to positively regulate root development.

Maintenance and homeostasis of the quiescent center (QC) in Arabidopsis (Arabidopsis thaliana) root apical meristems are critical for stem cell organization and root development. Despite great progress in relevant research, the molecular mechanisms that determine the root stem cell fate and QC still need further exploration. In Arabidopsis, SUPPRESSOR OF FRIGIDA 4 (SUF4) encodes a C2H2-type zinc finger protein that represses flowering by transcriptional activation of FLOWERING LOCUS C (FLC) through the FRIGIDA (FRI) pathway, and EARLY BOLTING IN SHORT DAYS (EBS) is a bivalent histone reader that prevents premature flowering. Here, we found that SUF4 directly interacts with EBS in vivo and in vitro. Loss of function of SUF4 and/or EBS resulted in disorganization of the QC, aberrant cell division, and stunted root growth. RNA-seq and reverse transcription quantitative real-time polymerase chain reaction analysis revealed that SUF4 and EBS coregulate many root development-related genes. A series of biochemical analyses demonstrated that SUF4 directly binds to the promoter of SCARECROW (SCR), which encodes a key regulator of root development. Chromatin immunoprecipitation assay indicated that both SUF4 and EBS are recruited to the SCR locus in an interdependent manner to promote H3K4me3 levels and suppress H3K27me3 levels, thereby activating the expression of SCR. These findings improve our understanding of the function of SUF4 and EBS and provide insights into the molecular mechanism that couples a transcription factor and a histone methylation reader to modulate QC specification and root development in Arabidopsis.

Deep Learning Image Recognition-Assisted Atomic Force Microscopy for Single-Cell Efficient Mechanics in Co-culture Environments.

Atomic force microscopy (AFM)-based force spectroscopy assay has become an important method for characterizing the mechanical properties of single living cells under aqueous conditions, but a disadvantage is its reliance on manual operation and experience as well as the resulting low throughput. Particularly, providing a capacity to accurately identify the type of the cell grown in co-culture environments without the need of fluorescent labeling will further facilitate the applications of AFM in life sciences. Here, we present a study of deep learning image recognition-assisted AFM, which not only enables fluorescence-independent recognition of the identity of single co-cultured cells but also allows efficient downstream AFM force measurements of the identified cells. With the use of the deep learning-based image recognition model, the viability and type of individual cells grown in co-culture environments were identified directly from the optical bright-field images, which were confirmed by the following cell growth and fluorescent labeling results. Based on the image recognition results, the positional relationship between the AFM probe and the targeted cell was automatically determined, allowing the precise movement of the AFM probe to the target cell to perform force measurements. The experimental results show that the presented method was applicable not only to the conventional (microsphere-modified) AFM probe used in AFM indentation assay for measuring the Young's modulus of single co-cultured cells but also to the single-cell probe used in AFM-based single-cell force spectroscopy (SCFS) assay for measuring the adhesion forces of single co-cultured cells. The study illustrates deep learning imaging recognition-assisted AFM as a promising approach for label-free and high-throughput detection of single-cell mechanics under co-culture conditions, which will facilitate unraveling the mechanical cues involved in cell-cell interactions in their native states at the single-cell level and will benefit the field of mechanobiology.

Boundary of mandibular molar distalization in orthodontic treatment: A systematic review and meta-analysis.

To explore the mandibular retromolar space length (MRSL), initial root-inner cortex contact percentage (IRCCP), and the various factors that influence mandibular molar distalization. Searches were undertaken in PubMed, EMBASE, Web of Science, Cochrane Library, Scopus, and grey literature (Google Scholar and OpenGrey) for eligible cross-sectional observational studies measuring the MRSL and IRCCP in healthy adult patients. The risk of bias and evidence quality were evaluated using the Joanna Briggs Institute's checklist and GRADE framework. Thirteen studies involving 1169 patients were included for qualitative synthesis. Seven of these studies were eligible for quantitative analysis. Meta-analysis showed that the mean MRSL at the subfurcation-6 mm plane in Asian normodivergent cases was 3.78 mm (95% confidence interval [CI]: 2.81-4.35; I2 = 79.7%) for skeletal Class-I malocclusions, 3.02 mm (95% CI: 2.10-3.94; I2 = 62.5%) for Class-II, and 4.43 mm (95% CI: 3.14-5.73; I2 = 75.1%) for Class-III. The mean MRSL at the sub-cementoenamel junction (CEJ)-10 mm plane for Asian, Class-I, normodivergent cases was 3.28 mm (95% CI: 2.44-4.12; I2 = 68.9%). The mean IRCCP for Asian, Class-I, normodivergent cases was 27.2% (95% CI: 0.22-0.32; I2 = 0%). In Asian normodivergent cases, MRSL ranges from 3.28 to 4.43 mm with a 27.2% IRCCP for Class-I. Cone-beam computed tomography imaging is recommended for measuring the MRSL in the apex region particularly before molar distalization. Factors influencing MRSL and IRCCP include different races, skeletal patterns, facial types, and third-molar status.

Association between occlusal features and masticatory function in Hong Kong preschool children: a survey with one-year longitudinal follow-up.

BACKGROUND: Mastication is important for breaking down food, aiding swallowing and nutrients absorption, and is therefore fundamental to a child's development. Studies have shown poor masticatory function to be associated with younger age and presence of caries. However, studies of the association between masticatory function and malocclusion yielded contradictory results. The aim of this study is therefore to investigate the association between three-dimensional occlusal features with masticatory function, among preschool children in Hong Kong. METHODS: Self-administered questionnaires on masticatory function in three domains, namely general chewing difficulty, requiring help when eating different food types and increased preference for soft food were completed by parents. Information on non-nutritive sucking habits and basic demographics were also collected in the questionnaire. Clinical examinations were conducted to record three-dimensional occlusal features and presence of caries. Baseline investigations and one-year follow-ups were undertaken for 1,566 and 996 preschool children. Association of poor masticatory function with occlusal features, sucking habits and caries was investigated using chi-squared tests. Binomial logistic regressions were then carried out incorporating any significant factors identified. Longitudinal analysis of the one-year follow-up data was carried out to investigate whether improved occlusal features, sucking habits and caries resulted in better masticatory function. RESULTS: In the cross-sectional study, the first domain of general chewing difficulty was associated with caries and thumb/digit sucking. The second domain of requiring help when eating different food types was associated with the male sex, younger age, caries and pacifier use. The last domain of increased preference for soft foods was associated with caries and thumb/digit sucking. Occlusal features, including abnormal overjet and unilateral permanent molars not in contact, were significantly associated with poor masticatory function in the bivariate analyses, but were not significant in the logistic regressions. In the longitudinal analysis, general chewing difficulty was found to improve in those of older age and those with resolved anterior crossbite. Less help was required to eat meat in those with fewer caries. Similarly, less help was required to eat food containing bones in those with reduced pacifier use. Preferences for eating soft foods was reduced in those who developed a normal overjet. CONCLUSIONS: The study identified significant relationships between masticatory difficulties and factors associated with age, gender, active caries, and non-nutritive oral habits such as thumb/digit sucking and pacifier use. Younger children and males required more assistance with certain food types. Active caries and thumb/digit sucking habits contributed to general masticatory difficulties and preference for soft foods. The one-year follow-up indicated that improvement in masticatory function varies across age cohorts and were associated with improved occlusal features, such as resolution of anterior crossbite and normalized overjet, reduced pacifier use, and a decrease in the number of decayed teeth.

Periodontal ligament cells from patients with treated stable periodontitis: Characterization and osteogenic differentiation potential.

BACKGROUND AND OBJECTIVE: Periodontal ligament progenitor cells (PDL cells) isolated from patients with inflammatory periodontitis have impaired regenerative capacity, but it is unknown whether this capacity can be recovered upon treatment and stabilization of the periodontal condition. The study aimed to investigate the expression of surface markers and the proliferation and osteogenic potential of PDL cells isolated from patients with treated stable periodontitis (S-PDL cells), periodontally healthy individuals (H-PDL cells), and patients with inflammatory periodontitis (I-PDL cells). METHODS: H-PDL, I-PDL, and S-PDL cells were isolated from the extracted teeth of individuals who (1) were periodontally healthy, (2) had inflammatory periodontitis, and (3) had treated stable periodontitis, respectively. The expression levels of surface markers and the proliferative and osteogenic capacities of the PDL cells were assessed. RESULTS: PDL cells derived from all three sources exhibited mesenchymal stem cell (MSC) characteristics. They were positive for MSC-related markers and negative for a hematopoiesis-related marker. However, S-PDL cells had higher proliferation rates, higher expression levels of osteogenic markers, higher alkaline phosphatase activity, and more calcium nodules than I-PDL cells. But all of these parameters remained lower in S-PDL cells than in H-PDL cells. CONCLUSIONS: S-PDL cells proliferated faster and had greater osteogenic potential than I-PDL cells, although these values remained lower than those in H-PDL cells.

Arabidopsis SEC13B Interacts with Suppressor of Frigida 4 to Repress Flowering.

SECRETORY13 (SEC13) is an essential member of the coat protein complex II (COPII), which was reported to mediate vesicular-specific transport from the endoplasmic reticulum (ER) to the Golgi apparatus and plays a crucial role in early secretory pathways. In Arabidopsis, there are two homologous proteins of SEC13: SEC13A and SEC13B. SUPPRESSOR OF FRIGIDA 4 (SUF4) encodes a C2H2-type zinc finger protein that inhibits flowering by transcriptionally activating the FLOWERING LOCUS C (FLC) through the FRIGIDA (FRI) pathway in Arabidopsis. However, it remains unclear whether SEC13 proteins are involved in Arabidopsis flowering. In this study, we first identified that the sec13b mutant exhibited early flowering under both long-day and short-day conditions. Quantitative real-time PCR (qRT-PCR) analysis showed that both SEC13A and SEC13B were expressed in all the checked tissues, and transient expression assays indicated that SEC13A and SEC13B were localized not only in the ER but also in the nucleus. Then, we identified that SEC13A and SEC13B could interact with SUF4 in vitro and in vivo. Interestingly, both sec13b and suf4 single mutants flowered earlier than the wild type (Col-0), whereas the sec13b suf4 double mutant flowered even earlier than all the others. In addition, the expression of flowering inhibitor FLC was down-regulated, and the expressions of flowering activator FLOWERING LOCUS T (FT), CONSTANS (CO), and SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) were up-regulated in sec13b, suf4, and sec13b suf4 mutants, compared with Col-0. Taken together, our results indicated that SEC13B interacted with SUF4, and they may co-regulate the same genes in flowering-regulation pathways. These results also suggested that the COPII component could function in flowering in Arabidopsis.

Pre-Epiglottic Baton Plate in the Management of Upper Airway Obstruction in an Infant with Femoral Facial Syndrome: A Case Report.

Femoral facial syndrome (FFS) is a rare condition which may present with hypoplasia or aplasia of the femora and unusual facies characterized by long philtrum, thin upper lip and micrognathia. We present the case of a ten-month old infant with FFS who had retroglossal obstruction and who was treated with a pre-epiglottic baton plate. The pre-epiglottic baton plate can be a simple, non-invasive and effective tool for the clinical management of syndromic patients with mild-to-moderate upper airway obstruction due to micrognathia.

Caries-preventive effectiveness of two different fluoride varnishes: A randomised clinical trial in patients with multi-bracketed fixed orthodontic appliances.

BACKGROUND: Patients with multi-bracketed fixed orthodontic appliances are at a higher risk of developing enamel demineralisation. AIM: To compare the efficacy of the quarterly application of two varnishes in preventing white spot lesions (WSLs) during multi-bracketed fixed orthodontic treatment. DESIGN: Ninety-nine patients were randomly allocated into three intervention groups: Group A-standard oral hygiene instructions (OHI); Group B-5% sodium fluoride (NaF) varnish with standard OHI; and Group C-NaF plus tricalcium phosphate varnish with standard OHI. The outcome was assessed at 6-monthly intervals using a quantitative laser fluorescence device, clinical evaluation and photographic evaluation on six maxillary anterior teeth. A logistic regression model using generalised estimating equations with the intervention group, time and their interaction was used to compare the occurrence of WSLs, whereas the independent-samples Kruskal-Wallis test was used to compare the means of fluorescent values among different groups at p = .05. RESULTS: Generalised estimating equations (with intervention and time as predictors) showed that only the effect of time was found to be significant (p < .001), whereas there was no significant difference among the three intervention groups (p = .305) using clinical assessment or photographic assessment (p = .599). At the 18-month follow-up, sodium fluoride (NaF) varnish with standard OHI was shown to reduce the odds of WSLs by 55% (p = .200), and NaF plus tricalcium phosphate varnish with standard OHI by 42% (p = .327). CONCLUSION: The study failed to demonstrate that the quarterly application of both the study varnishes with OHI provided additional benefits compared with standard OHI alone in preventing WSLs, taking the effect of time of follow-up into consideration. There were higher odds of developing WSLs with an increased duration of orthodontic treatment.

The reliability of analytical reference lines for determining esthetically pleasing lip position: An assessment of consistency, sensitivity, and specificity.

INTRODUCTION: This study aimed to identify a simple yet reliable soft-tissue parameter for the clinical determination of esthetic lip position by investigating the most consistent reference lines and assessing their sensitivity and specificity. METHODS: A total of 5745 records from Chinese patients aged >18 years were screened. In part I of the study, lateral view photographs of 96 subjects (33 males, 63 females) with esthetic facial profiles were selected. The profile esthetics of each photograph was first scored by 52 dental students, followed by 97 laypeople on a 5-point attractiveness scale. For the top 25% of photographs with the highest score for each sex (8 males, 16 females), the consistency of 6 commonly used reference lines were assessed to determine the esthetic lip position. In part II of the study, lip positions relative to Steiner's (S) and Ricketts' (E) lines in the profile photographs of 86 patients (43 males, 43 females) deemed to have an esthetically unpleasing profile were compared with those in 86 Chinese movie star idols (43 males, 43 females). RESULTS: In part I of the study, the S, E, and Burstone's (B) lines exhibited the lowest standard deviations for the upper and lower lips. B line was excluded from further analysis because of its higher mean absolute values, and S and E lines were used for the subjective assessment in part II of the study. In part II, the S line showed a sensitivity of 86.0% and 86.0% and a specificity of 81.4% and 83.7% for males and females, respectively. In contrast, the E line presented a sensitivity of 88.4% and 93.0% and a specificity of 79.1% and 74.4% for males and females, respectively. CONCLUSIONS: S, E, and B lines were the most consistent soft-tissue parameters among both sexes; however, because of the smaller absolute values, the S line would be more convenient among the 3 for a quick clinical assessment of lip position. Moreover, the performance of both S and E lines was similar among both sexes, which supports using these lines in assessing the esthetic lip position.

Minimally important differences in oral health-related quality of life after fixed orthodontic treatment: a prospective cohort study.

BACKGROUND: The minimally important difference (MID, the smallest difference that patients perceive as beneficial) has been proposed to assess whether study results are clinically meaningful, reducing the shortcoming of P-values-based approaches in the assessment of clinical outcomes. The post-treatment changes and the MID in Oral Health-Related Quality of Life (OHRQoL) among adults undergoing fixed orthodontic treatment were investigated. METHODS: Ninety-two patients (29.1 ±â€…6.3 years old, 18 males and 74 females) were prospectively included. Oral Health Impact Profile (OHIP-14) and United Kingdom Oral Health-related Quality of Life (OHQoL-UK) were used to assess OHRQoL at baseline and post-treatment (50.8 ±â€…15.7 months). Global Transition Rating (GTR) was used to assess oral health/well-being, Peer Assessment Review (PAR), and Index of Complexity, Outcome and Need (ICON) were used to assess occlusion. Wilcoxon signed-rank test was used to assess changes in OHRQoL and occlusion, Kruskal-Wallis one-way ANOVA and Mann-Whitney U-test were used to assess associations between OHRQoL and oral health/well-being or occlusion. MID for OHIP-14 and OHQoL-UK was estimated via anchor-based (PAR, ICON, GTR) and distribution-based approach (effect size, standardized response mean, standard error of measurement). RESULTS: The median OHIP-14 and OHQoL-UK post-treatment scores were significantly changed, indicating improved OHRQoL. Based on the agreement between different methods, the MID of OHIP-14 and OHQoL-UK were at least 15 and 6 scale points, corresponding to a large effect size (1.5-1.7). CONCLUSIONS: Orthodontic treatment had a positive long-term impact on OHRQoL. MID for the OHIP-14 and OHQoL-UK provide guidance to interpreting the impact of orthodontic treatment on the OHRQoL of patients.

Improvement of parents' oral health knowledge by a school-based oral health promotion for parents of preschool children: a prospective observational study.

BACKGROUND: Parents of preschool children have inadequate oral health knowledge in Hong Kong. Parents play a critical role in preschool children's dietary patterns and oral health behaviors. A school-based oral health promotion (OHP) for parents of preschoolers was developed and investigated. OBJECTIVES: The objective of this study was to evaluate effects of the school-based OHP for parents of preschool children on parents' oral health knowledge and preschool children's early childhood caries (ECC). MATERIALS AND METHODS: This was a quasi-experimental study. Parents of preschool children were divided into the intervention group (IG) and the control group (CG) according to their own selection. Parents in the IG participated in a structured school-based OHP workshop, while those in the CG did not attend the OHP workshop. Parents in both groups were invited to complete a questionnaire assessing their oral health knowledge before (T0), one month after (T1), and twelve months after (T2) the OHP workshop. Preschool children's caries was examined via dmft score at T0 and T2. RESULTS: Parents' oral health knowledge was negatively correlated with preschool children's dmft scores (R = -0.200, P < 0.001). Oral health knowledge was significantly improved in IG (P < 0.001) but not in CG (P = 0.392) at T1. Both groups experienced a significant improvement in oral health knowledge from T0 to T2 (P < 0.001). Parents' oral health knowledge in the IG was significantly higher compared to the CG at T1 (P < 0.001), but difference in the scores at T2 between the two groups showed no significant difference (P = 0.727). No significant difference was found in changes in children's dmft score from T0 to T2 between the IG and CG (p = 0.545). CONCLUSION: Preschool children's high ECC is associated with the limited oral health knowledge of their parents. The school-based OHP workshop for parents increased parents' oral health knowledge within one month. This positive effect was maintained for twelve months and can be extended to a larger scale in the school setting.

Nasal asymmetry changes during growth and development in 6- to 12-year-old children with repaired unilateral cleft lip and palate: A 3D computed tomography analysis.

Repaired unilateral cleft lip and palate (UCLP) is often accompanied by the deformity and asymmetry of the nasal region. Three-dimensional analysis was performed to investigate the relationship between nasal soft- and hard-tissue asymmetries, as well as the changes in nasal asymmetry with age, among children with repaired UCLP (age: 6-12 years). Forty-seven patients were included in this study. Their computed tomography records were retrieved for analysis of the 3D asymmetry of 10 landmarks of the nasal soft and hard tissues. We observed that asymmetry was more severe in nasal hard tissues than in soft tissues, particularly in the sagittal dimension. Compared with patients aged 6-9 years old, patients aged 10 to 12 years old had significantly increased vertical asymmetry at the base of the alar groove (Gbase, p = 0.027) and the lateral point of the piriform aperture (LPA), (p < 0.001). The correlation between the LPA and the alar region was weak to moderate (r = 0.290 to 0.488). In conclusion, we found no evidence of growth and development in nasal hard-tissue asymmetry among 6- to 12-year-old children with repaired UCLP, except for the vertical dimension. Nasal soft tissue exhibited a more preferable symmetry than hard tissue, and this could be attributed to the compensatory growth of nasal soft tissue, particularly in the vertical and sagittal dimensions. The weak to moderate correlations between nasal soft-tissue asymmetry and hard-tissue asymmetry were observed in the three dimensions. Surgeons should consider these factors when repositioning the nasal alar and controlling the size of the nostrils.

Lipopolysaccharides affect compressed periodontal ligament cells via Eph-ephrin signaling.

OBJECTIVES: The aim of this study is to investigate the underlying mechanism of the recovery of periodontal ligament cells (PDLCs) sequentially exposed to inflammation and mechanical loading. MATERIALS AND METHODS: We divided PDLCs into four groups: control; compressive force (CF) alone (2.0 g/cm2 ); lipopolysaccharides (LPS) pretreatment (0.1 µg/ml) followed by simultaneous LPS and CF stimulation, simulating uncontrolled periodontitis; and LPS pretreatment followed by CF exposure, simulating controlled periodontitis. The expression of EphB4-ephrinB2 and EphA2-ephrinA2, and the level of osteoclastogenesis and osteogenesis were evaluated. RESULTS: Simultaneous stimulation by LPS and CF, compared with CF alone and sequential LPS and CF exposure, significantly suppressed EphB4 and enhanced ephrinA2 expression. Similarly, the most intense osteoclastic differentiation was observed under simultaneous LPS and CF stimulation, while sequential exposure to LPS and CF only slightly increased osteoclastic cell numbers. Both the activation of EphB4 signaling and ephrinA2 silencing lowered osteoclastic differentiation, which had previously been upregulated by simultaneous LPS and CF stimulation. These treatments also increased osteogenic differentiation. CONCLUSIONS: Simultaneous LPS and CF stimulation critically enhances osteoclastogenesis in PDLCs through the suppression of EphB4 and the induction of ephrinA2 signaling. Sequential LPS and CF exposure partially abolishes the osteolytic effects of simultaneous stimulation.

Rno_circRNA_008646 regulates formaldehyde induced lung injury through Rno-miR-224 mediated FOXI1/CFTR axis.

Formaldehyde (FA) serves as a prevailing air pollutant, which has seriously threatened public health in recent years. Of all the known health effects, lung injury is one of the most severe risks. However, little is known about the circRNAs related molecular mechanism in the development of lung injury induced by FA. This study was designed to explore the potential roles of dysregulated circRNAs as well as its mechanism in FA-induced lung injury. In the present study, 24 male SD rats were exposed to formaldehyde (control, 0.5, 2.46 and 5 mg/m3) for 8 h per day for 8 weeks to induce lung injury. We used H&E staining to evaluate the histopathological changes of lung injury indifferent groups. The expression of circRNAs in lung tissue was detected by real-time PCR. Meanwhile, circRNA/miRNA/mRNA interaction networks were predicted by bioinformatics analysis. Our study revealed that formaldehyde exposure resulted in abnormal histopathological changes in lung tissues. Moreover, the expression of rno_circRNA_008646 was significantly higher in lung tissues of formaldehyde exposure rats than in control. Bioinformatics analysis showed that one potential target miRNA/mRNA for rno_circRNA_008646 was rno-miR-224/Forkhead Box I1 (FOXI1). Besides, luciferase report gene confirmed that there was targeted binding relationship between rno_circRNA_008646 and rno-miR-224, rno-miR-224 and FOXI1. Further verification experiments indicated that the expression of rno_circRNA_008646 was negatively correlated rno-miR-224, while it was positively correlated with FOXI1. JASPAR database showed transcription factor FOXI1 located in promotor of CF Transmembrane Conductance Regulator (CFTR). Both FOXI1 and CFTR were up-regulated in lung tissues after formaldehyde exposure. In conclusion, our findings suggested that formaldehyde may induce lung injury, and this may be caused by up-regulatedrno_circRNA_008646, which medicated rno-miR-224/FOXI1/CFTR axis.

Prevention of demineralization during multi-bracketed fixed orthodontic treatment: An overview of systematic reviews.

BACKGROUND: Demineralization during multi-bracketed fixed orthodontic treatment is a well-known problem. AIM: To systematically evaluate the evidence for the prevention of enamel demineralization during multi-bracketed fixed orthodontic treatment. DESIGN: Systematic reviews (with or without meta-analysis) that have appraised the primary studies on the prevention of demineralization during multi-bracketed fixed orthodontic treatment were searched systematically and included as per pre-defined eligibility criteria. The risk of bias of the included reviews was assessed by two authors using the ROBIS tool and AMSTAR-2 tool. RESULTS: Twenty-nine reviews conducted on 128 exclusive primary studies were included for the present overview; 23 of these reviews were assessed to be of a high risk of overall bias, 5 were of low risk, and 1 review was of unclear risk of bias. CONCLUSIONS: Predominantly, the published reviews have focused on fluorides, whereas some reviews have also studied the role of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), reminder therapy, lasers, and sealants. Professional fluorides, sealants, reminder therapy, and lasers might be valuable interventions to prevent demineralization during orthodontic treatment based on the conclusions of reviews with high quality; however, further studies are required to elucidate the role of CPP-ACP, chlorhexidine varnish, and powered toothbrushes.

3-Dimensional simulations and student learning in orthodontic education.

INTRODUCTION: The electronic dental model (e-model) is an example of a digital 3-dimensional technology to support inquiry-based learning in undergraduate dental education. As student perceptions of and engagement with e-models vary, it is uncertain whether these perceptions have implications for their learning processes and outcomes. MATERIALS AND METHODS: Third-year dental students (N = 40) completed a questionnaire to identify their perceptions of and preferences for model modalities. They were divided into three groups based on their preference: Preferring plaster models (Group 1); Preferring e-models (Group 2); No preference (Group 3). Students from three groups (N = 9) attended a hands-on digital occlusion evaluation workshop, and then completed a case-based diagnostic evaluation test using digital occlusion evaluation software. Camtasia Studio™ recorded real-time and on-screen data of the number of mouse-clicks and time spent. RESULTS: Students reported positive feedbacks on the use of e-models, and 72.5% of the students preferred combination use of e-models and plaster models. After attending the hands-on digital dental occlusion evaluation workshop, Group 2 scored higher on the diagnostic evaluation test (p < .05) and registered more mouse-clicks than Group 1 when evaluating the arch symmetry (p < .05). Group 2 registered fewer mouse-clicks than Group 3 during tooth size measurement (p < .05). There was no significant difference regarding the time used to answer the knowledge questions amongst the three groups. CONCLUSION: Undergraduate dental students indicated a generally high acceptance of e-models for their learning in orthodontics, and more prefer a blended approach. Students preferring e-models presented higher performance outcomes, which supports cognitive load theory regarding prior exposure to simulation-based environments.

Temporal trends in semen concentration and count among 327 373 Chinese healthy men from 1981 to 2019: a systematic review.

STUDY QUESTION: Are there temporal trends of sperm concentration (SC) and total sperm count (TSC) in Chinese healthy males from 1981 to 2019? SUMMARY ANSWER: Our result indicated a temporal decrease in SC and TSC among 327 373 healthy Chinese men in the recent four decades. WHAT IS KNOWN ALREADY: A review of 61 papers reported a temporal decline in SC and TSC from 1938 to 1990. This trend was later confirmed by a systematic review of 185 published papers from 1981 to 2013. However, the majority of the included individuals were from western countries. In China, whether SC and TSC have declined remains controversial. STUDY DESIGN, SIZE, DURATION: This systematic review of published articles used data extracted from Pubmed, Science Direct, Embase, China-National-Knowledge-Infrastructure (CNKI) and Wanfang Data to assess changes in SC and TSC in China from 1981 to 2019. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 111 studies including 327 373 individuals who provided semen samples from 1981 to 2019 were extracted for the present analysis. Study selection and data extraction were performed by two independent researchers. The trends in SC and TSC were analysed using liner-regression and meta-regression before and after adjusting for potential covariates. Moreover, subgroups, categorised based on geographic region, fertility status or recruitment source, were also analysed. MAIN RESULTS AND THE ROLE OF CHANCE: SC declined significantly (slope liner-regression = -0.748 million/ml/year; P = 0.005; slope meta-regression = -0.824 million/ml/year; P < 0.001) between 1981 and 2019 in China. Trends for TSC was similar to that for SC (slope liner-regression = -2.073 million/year; P = 0.032; slope meta-regression = -2.188 million/year; P = 0.003). In subgroup meta-regression analyses, males with definite fertility had continuous declines in SC (slope northern group=-2.268, P = 0.009; slope southern group=-1.014, P = 0.009) and TSC (slope northern group=-9.675, P = 0.010; slope southern group=-3.215, P = 0.042). However, in the unselected group, where fertility status was unknown, the obvious downward trend in SC was only seen in males from Northern regions (slope = -0.836, P = 0.003). Another subgroup analysis demonstrated that obvious decreases in SC (slope = -1.432, P < 0.001) and TSC (slope=-4.315, P = 0.001) were only seen in volunteer groups but not in pre-pregnancy examination groups and other recruitment groups. The results changed minimally in multiple sensitivity analyses. LIMITATIONS, REASONS FOR CAUTION: The validity of the meta-analysis results was limited mainly by the quality of the included studies. Additionally, our study spanned many decades and the recommended criteria for some semen parameter assessments have significantly changed, which may bring about some unavoidable bias. Moreover, the data remain insufficient especially in some provinces of China. WIDER IMPLICATIONS OF THE FINDINGS: The present study is the first study to report significant decreases in SC and TSC in 327 373 healthy Chinese men between 1981 and 2019, indicating a serious reproductive health warning. Further studies on the causes of the declines are urgently needed. STUDY FUNDING/COMPETING INTEREST(S): D.Z. is supported by the National Natural Science Funding of China, Natural Science Funding of Shaanxi Province, Science Funding of Health Department, Shaanxi Province, Fundamental Research Funds for the Central University and the Project of Independent Innovative Experiment for Postgraduates in Medicine in Xi'an Jiaotong University. The authors have no conflicts of interests to declare. TRIAL REGISTRATION NUMBER: N/A.

Identification and integrated analysis of differentially expressed long non-coding RNAs associated with periodontitis in humans.

BACKGROUND AND OBJECTIVE: Long non-coding RNAs (lncRNAs) can act as competing endogenous RNAs (ceRNAs) to compete for micro-RNAs (miRNAs) in regulation of downstream genes, various biological functions and diseases. Yet, the expression and regulation of lncRNAs in periodontitis are not fully understood. The objective of the study was to identify potential genes (lncRNA, messenger RNA [mRNA] and miRNA) involved in periodontitis, construct lncRNA-miRNA-mRNA ceRNA networks, explore gene functions and validate gene expressions. MATERIAL AND METHODS: The data sets for the lncRNA, mRNA and miRNA expression profiles in gingival samples from periodontally healthy subjects and chronic periodontitis patients were obtained from the Gene Expression Omnibus. The differentially expressed lncRNAs (DElncRNAs), mRNAs (DEmRNAs) and miRNAs (DEmiRNAs) were identified, and ceRNA networks were then constructed. The expression of DElncRNAs and DEmRNAs was examined by quantitative real-time polymerase chain reaction (qPCR). Moreover, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed for exploring the potential functions and biological pathways. RESULTS: The GSE80715 and GSE54710 data sets were retrieved. Subsequently, 26 DElncRNAs, 436 DEmRNAs and 12 DEmiRNAs were identified (|fold change| ≥2, adjusted p < 0.05). Further bioinformatics analysis contributed to establishment of the ceRNA networks, which consisted of 10 DElncRNAs, 11 DEmiRNAs and 83 DEmRNAs. Notably, the qPCR results showed a marked decrease in the expression of lncRNA H19 and two mRNAs (NOS1 and MAPT) which further supported the identified ceRNA network. The GO results revealed that the up-regulated mRNAs were significantly enriched in inflammatory processes, whilst the down-regulated mRNAs were enriched in cellular potentials. CONCLUSION: Non-coding RNAs are critically involved in the regulatory mechanisms in the pathogenesis of periodontitis. Further study is warranted to investigate the specific underlying genetic traits and networks.

Long-term exposure to formaldehyde induced down-regulation of SPO11 in rats.

Objective: Formaldehyde, a ubiquitous environmental contaminant, has long been suspected of causing male reproductive injury, but the underlying molecular mechanism remains largely unknown. SPO11 is a meiosis-related gene, whose absence can cause spermatogenesis arrest. Materials and methods: The present study aimed to explore the role of SPO11 in male reproductive injury induced by long-term formaldehyde exposure, so as to further understand the molecular mechanism of formaldehyde-induced male reproductive toxicity. Adult male Sprague-Dawley rats (n = 24, 245 ± 22 g) were randomly divided into four groups of six (n = 6) and were exposed to formaldehyde gas at doses of 0 (control), 0.5, 2.46 and 5 mg/m3, respectively, via inhalation for 8 consecutive weeks. Results and dissussion: The expression levels of SPO11 were detected in testicular tissues by real-time quantitative polymerase chain reaction, immunofluorescence, and Western blot. The results indicated that the expression of SPO11 was inhibited by formaldehyde exposure in a dose-dependent manner. Furthermore, the histopathological results showed that testicular seminiferous tubules were atrophied, spermatogenic cells were decreased and the lumina were oligozoospermic in the 2.46 and 5 mg/m3 formaldehyde exposure groups. Combined with the morphometric results, we found that the downregulated expression levels of SPO11 were consistent with the changes of testicular seminiferous tubule diameter and seminiferous epithelium height in testicular tissue, suggesting that SPO11 might be one of the main targets of formaldehyde reproductive toxicity. Conclusions: In conclusion, our findings indicated that SPO11 might be related to male reproductive injuries induced by long-term formaldehyde exposure.

Circular RNA-CDR1as is involved in lung injury induced by long-term formaldehyde inhalation.

OBJECTIVE: Formaldehyde (FA) is known to induce lung injury, but the underlying molecular mechanism remains largely unclear. CDR1as is an important member of the circular RNAs (circRNAs) family and functions as miRNA sponges with gene-regulatory potential. Our earlier circRNA microarray data showed CDR1as was highly expressed in lung tissue exposed to FA. However, the mechanism of circRNA-CDR1as mediates the FA-exposed lung injury is still unclear. This study aimed to explore the role of CDR1as in lung injury. MATERIALS AND METHODS: In this study, FA was inhaled at doses of 0.5, 2.46, and 5 mg/m3, respectively. After exposure 8 weeks, lung histopathological examination, lung injury score, and IL-1ß in bronchoalveolar lavage fluid (BALF) were determined. The expressions of CDR1as, rno-miR-7b and Atg7 were detected and the potential interaction of circRNA/miRNA/mRNA was predicted by bioinformatics analysis, including drawing circRNA/miRNA/mRNA interaction network, GO and KEGG analysis. RESULTS: Our results indicated FA inhalation upregulated the expression of CDR1as in lung tissues in a dose-dependent manner while the expression of rno-miR-7b decreased and Atg7 increased. Moreover, the alteration of CDR1as was positively correlated with lung injury. DISCUSSION AND CONCLUSIONS: CircRNA/miRNA/mRNA prediction further explained the possible effect mechanisms of CDR1as. These data implicated that CDR1as might be a critical regulator involved in lung injury induced by FA.