How to make full use of dental pulp stem cells: an optimized cell culture method based on explant technology.

Introduction: Dental pulp stem cells from humans possess self-renewal and versatile differentiation abilities. These cells, known as DPSC, are promising for tissue engineering due to their outstanding biological characteristics and ease of access without significant donor site trauma. Existing methods for isolating DPSC mainly include enzyme digestion and explant techniques. Compared with the enzymatic digestion technique, the outgrowth method is less prone to cell damage and loss during the operation, which is essential for DPSC with fewer tissue sources. Methods: In order to maximize the amount of stem cells harvested while reducing the cost of DPSC culture, the feasibility of the optimized explant technique was evaluated in this experiment. Cell morphology, minimum cell emergence time, the total amount of cells harvested, cell survival, and proliferative and differentiation capacity of DPSC obtained with different numbers of explant attachments (A1-A5) were evaluated. Results: There was a reduction in the survival rate of the cells in groups A2-A5, and the amount of harvested DPSC decreased in A3-A5 groups, but the DPSC harvested in groups A1-A4 had similar proliferative and differentiation abilities. However, starting from group A5, the survival rate, proliferation and differentiation ability of DPSC decreased significantly, and the adipogenic trend of the cells became more apparent, indicating that the cells had begun to enter the senescence state. Discussion: The results of our study demonstrated that the DPSC obtained by the optimized explant method up to 4 times had reliable biological properties and is available for tissue engineering.

The associations of two novel inflammation indexes, systemic immune-inflammation index (SII) and system inflammation response index (SIRI), with periodontitis: evidence from NHANES 2009-2014.

OBJECTIVES: The research's goal is to look for any potential relationships between the systemic immune-inflammation index (SII) and the system inflammation response index (SIRI), along with inflammation indicators and the likelihood of periodontitis. METHODS: Ten thousand two hundred eighty-two individuals in sum were determined to be eligible for this cross-sectional study from the National Health and Nutrition Examination Survey (NHANES) between 2009 and 2014. Multiple logistic regression, generalized additive model, smooth curve fitting, subgroup analysis, and interaction tests were done for analyzing the association between periodontitis and SII, SIRI, and other inflammatory indicators. RESULTS: The analysis, adjusted for population weighting, revealed that individuals with moderate/severe periodontitis had SII levels of 545.46 (95% CI (529.10, 561.82), P = 0.0044) and SIRI levels of 1.33 (95% CI (1.29, 1.37), P < 0.0001). In a fully adjusted multivariate logistic regression model, SII was not sensibly associated with moderate/severe periodontitis among the continuous and quartile Q1-Q4 groups (OR = 0.97, 95% CI (0.91, 1.02)). The continuous variable of SIRI (OR = 1.11, 95% CI (1.06, 1.17)) and the quartile Q4 group (OR = 1.58, 95% CI (1.28, 1.94)) had a deemed significant positive association with moderate to severe periodontitis. In addition, other inflammatory indicators, especially NLR, PPN, PLR, MLR, PC, NC, and MC were observed to be notably involved moderate/severe periodontist in this research. CONCLUSION: We explored the association between periodontitis and two novel comprehensive markers of inflammation (SII and SIRI). CLINICAL RELEVANCE: These inflammatory markers are expected to serve as tools to assist clinicians in diagnosing periodontitis.

Association between life's essential 8 and periodontitis: a population-based study.

BACKGROUND: The American Heart Association has developed a novel cardiovascular health indicator called Life's Essential 8 (LE8). However, no one has reported using LE8 to assess periodontitis. This study aimed to investigate the association between LE8 and periodontitis in American adults. METHODS: Data from the National Health and Nutrition Examination Survey (NHANES) from 2009 to 2014 were used for this investigation. LE8 was the independent variable, and it is divided into three grades: high, moderate, and low. Periodontitis was the dependent variable, and the classification of periodontitis was based on the criteria of Eke in 2012. Multivariable logistic regression models were used to explore the relationship between LE8 and periodontitis. RESULTS: A total of 9,039 participants with an average age of 52.16 ± 14.21 years were enrolled in this study, of whom 48.29% were male and 51.71% were female. The mean and standard deviation of LE8 was 66.29 ± 14.57, and the prevalence of periodontitis was 50.48% overall. The LE8 score and periodontitis in the fully adjusted logistic regression model showed a negative correlation (OR = 0.98; 95% CI, 0.98-0.99, p < 0.001). This result persisted when Life's Essential 8 was categorized into low, moderate, and high groups. Compared with those in the lowest group, those in the highest LE8 group had a 47% decreased risk of periodontitis (OR = 0.53; 95% CI, 0.46-0.66, P < 0.001). CONCLUSIONS: This cross-sectional investigation revealed a negative relationship between the LE8 score and the likelihood of periodontitis.

Clinical application of lithium disilicate glass ceramic cantilever resin-bonded fixed partial denture in the restoration of single anterior tooth loss / 口腔疾病防治

Objective@#To investigate the clinical effect of lithium disilicate glass ceramic cantilever resin-bonded fixed partial dentures (CRBFPDs) on single anterior tooth loss to provide a reference for the selection of restoration methods for single anterior tooth loss.@*Methods@#This study was reviewed and approved by the Ethics Committee, and informed consent was obtained from the patients. Forty-two patients with less than two anterior teeth with monomaxillary loss were included in this study. After 6 months, 1 year, 2 years, and 3 years, the aesthetic and functional effects of the restorations and the periodontal health status were evaluated, and the visual analog scale (VAS) was used to assess patient satisfaction.@*Results@#During the observation period, the connector fractured in one case within 3 months. One case had debonded within 2 years. The aesthetic restoration effect of all lithium disilicate glass ceramic CRBFPDs was categorized as Class A. The periodontal health was good, there was no clinical absorption in the soft and hard tissues of the abutment or subbridge, periodontal status according to the evaluation indices was classified as class A, and the total satisfaction rate of the patient was 100%.@*Conclusion@#For single anterior tooth loss patients, lithium disilicate glass ceramic cantilever resin-bonded fixed partial denture can achieve the restoration effect of less invasion, better adhesion, aesthetics, comfort and good biocompatibility. With high patient satisfaction, it can be considered an ideal restoration method for replacing a single anterior tooth.

Effect of tetracycline hydrochloride application on dental pulp stem cell metabolism-booster or obstacle for tissue engineering?

Introduction: Stem cells and scaffolds are an important foundation and starting point for tissue engineering. Human dental pulp stem cells (DPSC) are mesenchymal stem cells with self-renewal and multi-directional differentiation potential, and are ideal candidates for tissue engineering due to their excellent biological properties and accessibility without causing major trauma at the donor site. Tetracycline hydrochloride (TCH), a broad-spectrum antibiotic, has been widely used in recent years for the synthesis of cellular scaffolds to reduce the incidence of postoperative infections. Methods: In order to evaluate the effects of TCH on DPSC, the metabolism of DPSC in different concentrations of TCH environment was tested. Moreover, cell morphology, survival rates, proliferation rates, cell migration rates and differentiation abilities of DPSC at TCH concentrations of 0-500 µg/ml were measured. Phalloidin staining, live-dead staining, MTS assay, cell scratch assay and real-time PCR techniques were used to detect the changes in DPSC under varies TCH concentrations. Results: At TCH concentrations higher than 250 µg/ml, DPSC cells were sequestered, the proportion of dead cells increased, and the cell proliferation capacity and cell migration capacity decreased. The osteogenic and adipogenic differentiation abilities of DPSC, however, were already inhibited at TCH con-centrations higher than 50 µg/ml. Here, the expression of the osteogenic genes, runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN), the lipogenic genes lipase (LPL), as well as the peroxisome proliferator-activated receptor-γ (PPAR-γ) expression were found to be down-regulated. Discussion: The results of the study indicated that TCH in concentrations above 50 µg/ml negatively affects the differentiation capability of DPSC. In addition, TCH at concentrations above 250 µg/ml adversely affects the growth status, percentage of living cells, proliferation and migration ability of cells.

Analysis of 5-Year-old children's oral health service utilization and influencing factors in Guizhou Province, China (2019-2020).

BACKGROUND: This study aimed to investigate the utilization patterns and factors related to oral health care for 5-year-old preschoolers based on Andersen's Behavioural Model in Guizhou Province, Western China. METHOD: A cross-sectional study of 4,862 5-year-old preschoolers in 66 kindergartens was conducted in 2019 and 2020. A basic oral examination and a survey of parents and grandparents were conducted to gather data on oral health services. The results were analysed using chi-square tests and logistic regression analysis. RESULT: The utilization rate of oral health services for children in Guizhou province was 20.5%. The dmft was 4.43, and the rate of caries was 72.2%. The average cost of a dental visit was higher in rural areas and higher for girls. Logistic regression analysis revealed that dmft ≥ 6 teeth, a history of toothache, starting toothbrushing at age ≤ 3 years and limited parental knowledge were the primary factors impacting dental visits. CONCLUSION: Needs factors such as severe oral conditions and pain in children are the main reasons for the utilization of these services. This study underscores the urgency to actively promote the importance of oral health and expand insurance coverage for oral health services.

Association between C-Reactive protein and periodontitis in an obese population from the NHANES 2009-2010.

BACKGROUND: Various data have been obtained on the relationship between body mass index (BMI) and C-reactive protein (CRP) and periodontitis. The aim of this study was to determine whether CRP/BMI are associated with periodontitis using data from the National Health and Nutrition Examination Survey (NHANES) database. METHODS: A cross-sectional analysis of data from 3602 participants in the 2009-2010 NHANES cycle was performed. The definition of periodontitis was used to divide participants into four groups according to the criteria of Eke. Correlations between CRP/BMI and periodontitis were tested for statistical significance by means of descriptive statistics, multivariate regression, and subgroup-stratified analyses, with and without adjustments for confounders (such as age and sex). RESULTS: There were no statistically significant differences (p > 0.05) regarding BMI and the development of periodontitis. After adjustment for age, sex, race, marital status, annual family income, alcohol consumption, hypertension, smoking, chronic pulmonary disease, cardiovascular disease, diabetes, flossing, and arthritis, CRP correlated significantly with the development of periodontitis in the subgroups stratified by obesity, with an odds ratio (OR) of 1.2 (95% CI, 1.0 to 1.5). CONCLUSION: Through data analysis, we found an association between CRP levels and periodontitis prevalence in the American population, although this association was only present in the obese population. While there are several hypotheses about the underlying mechanism, further studies are needed to validate these findings.

An imConvNet-based deep learning model for Chinese medical named entity recognition.

BACKGROUND: With the development of current medical technology, information management becomes perfect in the medical field. Medical big data analysis is based on a large amount of medical and health data stored in the electronic medical system, such as electronic medical records and medical reports. How to fully exploit the resources of information included in these medical data has always been the subject of research by many scholars. The basis for text mining is named entity recognition (NER), which has its particularities in the medical field, where issues such as inadequate text resources and a large number of professional domain terms continue to face significant challenges in medical NER. METHODS: We improved the convolutional neural network model (imConvNet) to obtain additional text features. Concurrently, we continue to use the classical Bert pre-training model and BiLSTM model for named entity recognition. We use imConvNet model to extract additional word vector features and improve named entity recognition accuracy. The proposed model, named BERT-imConvNet-BiLSTM-CRF, is composed of four layers: BERT embedding layer-getting word embedding vector; imConvNet layer-capturing the context feature of each character; BiLSTM (Bidirectional Long Short-Term Memory) layer-capturing the long-distance dependencies; CRF (Conditional Random Field) layer-labeling characters based on their features and transfer rules. RESULTS: The average F1 score on the public medical data set yidu-s4k reached 91.38% when combined with the classical model; when real electronic medical record text in impacted wisdom teeth is used as the experimental object, the model's F1 score is 93.89%. They all show better results than classical models. CONCLUSIONS: The suggested novel model (imConvNet) significantly improves the recognition accuracy of Chinese medical named entities and applies to various medical corpora.

Dental Caries and Associated Factors in 3-5-Year-Old Children in Guizhou Province, China: An Epidemiological Survey (2015-2016).

Objective: This study aimed to explore the factors influencing dental caries among 3-5-year-old children in Guizhou Province and the interrelationship between these factors using structural equation modeling, while providing theoretical references to improve the prevention and control strategy. Materials and Methods: A total of 1,291 children aged 3-5 years in Guizhou Province were selected by a multistage stratified and whole group random sampling to examine the caries prevalence in whole-mouth deciduous teeth crowns, and parents were surveyed with questionnaires to analyze the caries-related factors. IBM SPSS Statistics v 23.0 software (IBM, Armonk, NY, USA) was used for statistical analysis. Results: The caries prevalence of children aged 3-5 years in Guizhou Province was 63.1%, the mean decayed-missing-filled teeth was 3.32, the caries filling rate was 0.5%, and there was no statistically significant difference between urban and rural areas and among genders in each age group; results of logistic regression analysis showed that the caries risk increased with the following factors: age, brushing frequency <2 times per day when parents did not take their children to the dentist, and with parents poor evaluation of the oral condition of their children. The higher the education of the parent, the lower the risk of children suffering from caries in deciduous teeth. Conclusions: With an overall poor situation about oral hygiene habits, oral healthcare attitude of the parents, and behavior transformation, the prevalence of dental caries in the deciduous teeth of children aged 3-5 years in Guizhou Province is high, and their caries status was severe, with more than 99% of the caries cases that were untreated. Therefore, prevention and treatment measures of caries in preschool children need strengthening through the improvement of public awareness and the enhancement of the management of oral health habits of their children.

Absorption Characteristics and Environmental Significance of Dissolved Organic Matter in Lake Dongping.

As an important water source in Shandong Province, Lake Dongping is one of the most important inland lakes to South-to-North Water Diversion East Route Project. We tried to provide the basis for real-time monitoring of water quality mutation by using CDOM absorption characteristics. The average values of CDOM absorption coefficients [a(280)，a(350)，a(440)] were (12.90±1.17)，(3.11±0.40) and (0.65±0.09) m-1. a(440) had linear relationships with total nitrogen (p<0.01), total phosphorus (p<0.001), dissolved organic carbon (p<0.001), chemical oxygen demand (p<0.001), Chlorophyll a (p<0.001), which can be used to estimate the water quality parameters. The results showed that CDOM absorption coefficient reflects the nutritional status of Lake Dongping, which is close to eutrophication level. There was an obvious decrease of CDOM absorption coefficients from river mouth of Dawen River to lake center and then to the outlets of lake. This illustrated that terrestrial input is the main source of nutrients and pollutants in Lake Dongping.

Cloning, characterization, expression, and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab, Eriocheir sinensis.

A full-length metallothionein-1(MT-1) cDNA was cloned from the Chinese mitten crab, Eriocheir sinensis, based upon the hepatopancreas cDNA library. The full-length cDNA contained a single 180 bp open reading frame that encoded a 59 amino acid protein. The deduced amino acid sequence was cysteine (Cys)-rich, with residues observed in patterns characteristic of other reported MTs: Cys-X-Cys, Cys-X-X-Cys, or Cys-X-X-X-Cys. Gene structure obtained via PCR yielded a 3816 bp gene, which was comprised of three exons and two introns arranged in a "3 + 2" pattern. The cloned 5'flanking region (1,735 bp) contained several predicted binding sites, which included MREs, AP-1, SP1, USF, GATA, HNF-1, and HSF. MT-1 mRNA expression analysis revealed that while levels were highest in the hepatopancreas, expression was abundant in testis and thoracic ganglia, moderate in intestine (P<0.05), and weak in other tissues (P<0.05). MT-1 mRNA expression exhibited reproductive variation in the male, with levels approximately tenfold greater in August, during seasonal gonadal maturation, compared to other times of the year. Cu2+ exposure via tank water (0-1 mg/l for 7 days) resulted in a dose-dependent bell curve response in MT-1 mRNA expression, with peak expression observed after exposure to 0.1 mg/l Cu2+. A time course experiment (0.1 mg/l Cu2+ over 9 days) revealed MT-1 mRNA expression peaked sharply on day 5 before gradually decreasing with prolonged exposure. In the present report, we provide sequence analysis of the first MT-1 gene cloned in E. sinensis, and evidence that its physiological and toxicological regulation is evolutionary conserved.

Molecular cloning and tissue expression of the fatty acid-binding protein (Es-FABP) gene in female Chinese mitten crab (Eriocheir sinensis).

BACKGROUND: Fatty acid-binding proteins (FABPs), small cytosolic proteins that function in the uptake and utilization of fatty acids, have been extensively studied in higher vertebrates while invertebrates have received little attention despite similar nutritional requirements during periods of reproductive activity. RESULTS: Therefore, a cDNA encoding Eriocheir sinensis FABP (Es-FABP) was cloned based upon EST analysis of a hepatopancreas cDNA library. The full length cDNA was 750 bp and encoded a 131 aa polypeptide that was highly homologous to related genes reported in shrimp. The 9108 bp Es-FABP gene contained four exons that were interrupted by three introns, a genomic organization common among FABP multigene family members in vertebrates. Gene expression analysis, as determined by RT-PCR, revealed the presence of Es-FABP transcripts in hepatopancreas, hemocytes, ovary, gills, muscle, thoracic ganglia, heart, and intestine, but not stomach or eyestalk. Real-time quantitative RT-PCR analysis revealed that Es-FABP expression in ovary, hemocytes, and hepatopancreas was dependent on the status of ovarian development, with peak expression observed in January. CONCLUSIONS: Evidence provided in the present report supports a role of Es-FABP in lipid transport during the period of rapid ovarian growth in E. sinensis, and indirectly confirms the participation of the hepatopancreas, ovary, and hemocytes in lipid nutrient absorption and utilization processes.

Molecular cloning and gene expression analysis of the leptin receptor in the Chinese mitten crab Eriocheir sinensis.

BACKGROUND: Leptin is an adipocyte-derived hormone with multiple functions that regulates energy homeostasis and reproductive functions. Increased knowledge of leptin receptor function will enhance our understanding of the physiological roles of leptin in animals. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, a full-length leptin receptor (lepr) cDNA, consisting of 1,353 nucleotides, was cloned from Chinese mitten crab (Eriocheir sinensis) using rapid amplification of cDNA ends (RACE) following the identification of a single expressed sequence tag (EST) clone in a cDNA library. The lepr cDNA consisted of a 22-nucleotide 5'-untranslated region (5' UTR), a 402-nucleotide open reading frame (ORF) and a 929-nucleotide 3' UTR. Multiple sequence alignments revealed that Chinese mitten crab lepr shared a conserved vacuolar protein sorting 55 (Vps55) domain with other species. Chinese mitten crab lepr expression was determined in various tissues and at three different reproductive stages using quantitative real-time RT-PCR. Lepr expression was highest in the intestine, thoracic ganglia, gonad, and accessory gonad, moderate in hepatopancreas and cranial ganglia, and low in muscle, gill, heart, haemocytes, and stomach. Furthermore, lepr expression was significantly higher in the intestine, gonad and thoracic ganglia in immature crabs relative to precocious and mature crabs. In contrast, lepr expression was significantly lower in the hepatopancreas of immature crabs relative to mature crabs. CONCLUSIONS/SIGNIFICANCE: We are the first to identify the lepr gene and to determine its gene expression patterns in various tissues and at three different reproductive stages in Chinese mitten crab. Taken together, our results suggest that lepr may be involved in the nutritional regulation of metabolism and reproduction in Chinese mitten crabs.