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Objective: Single-cell RNA sequencing (scRNA-seq) was used to analyze the developing mouse molars, in order to construct a spatiotemporal development atlas of pulp cells, and further to reveal the developmental process and regulatory mechanism of tooth development. Methods: Ten mandibular first molars from C57BL/6 mice in postnatal day (PN) 0 and 3 were respectively dissected and digested to obtain single-cell suspensions. scRNA-seq was performed on 10× Genomics platform. PN 7 mouse molar scRNA-seq data were obtained from our previous study. PN 0, 3, and 7 scRNA-seq data were integrated for following analysis. The initial quality control, mapping and single cell expression matrix construction were performed by Cell Ranger. Quality control, standardization, dimensional reduction and cluster analysis were performed by using Seurat. Monocle was used to generate the pseudotime trajectory. Scillus was used to perform gene ontology analysis. In order to detect the spatiotemporal change of different population of pulp cells, the marker genes of each cluster were demonstrated by RNAscope in situ hybridization. Results: There were twenty-six cell clusters within mouse molars, which were identified as eight different cell types, including dental pulp cells, dental follicle cells, epithelial cells, immune cells, endothelial cells, perivascular cells, glial cells and erythrocytes. We further re-clustered and analyzed dental pulp cells. Cluster 0 were mature pulp cells, which located at the upper portion of crown. The main functions of cluster 0 were osteogenesis and extracellular structure organization. Cluster 1 were apical papilla cells, which located at the apical part of roots, whose main functions were extracellular structure organization and organ development. Cluster 2 were cycling cells, which were actively proliferated, resided in the lower portion of the crown. Cluster 3 and 4 were preodontoblasts and odontoblasts, respectively. Their functions were closely related to biomineralization. The proportion of mature pulp cells increased with the development process, while the proportion of cycling cells and odontoblast lineage decreased. According to the expression pattern of marker genes of each cluster, we constructed a cell atlas of dental pulp. Pseudotime trajectory analysis found there were two development trajectories within dental pulp. They both started from SPARC related modular calcium binding 2 (Smoc2)+ dental papilla cells, then went through DNA topoisomerase Ⅱ alpha (Top2a)+ cycling cells, and finally divided into coxsackie virus and adenovirus receptor (Cxadr)+ mature pulp cells or dentin sialophosphoprotein (Dspp)+ odontoblasts two lineages. Conclusions: scRNA-seq could fully discover the intercellular heterogeneity of cells on transcriptome level, which provides a powerful tool to study the process and regulatory mechanism of organ development.
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OBJECTIVE@#To evaluate the clinical effect of root canal obturation therapy using cold flowable gutta-percha on young permanent teeth after apexification.@*METHODS@#Ninety cases of young permanent teeth with pulp necrosis or periapical periodontitis treated by apexification were randomly divided into two groups. The cases in each group were divided into single root canal and multiple root canal according to the number of the root canal, and divided into classifications Ⅰ, and Ⅱ/Ⅲ/Ⅳ according to Frank's classification of root development after apexification. Cold flowable gutta-percha and warm gutta-percha obturation techniques were used for root canal obturation of the two groups. The operation time was recorded, and the patients' therapy pain degree was evaluated by visual analog scale (VAS) immediately after operation. Periapical X-ray was performed after operation to evaluate the effect of the root canal filling. The total length of the root was divided into equal three parts on the X-ray film, and three-dimensional tightness of the apical, middle, and coronal region of the root canals were statistically analyzed respectively. Clinical examinations and X-ray examination were performed 6 and 12 months after the operation to evaluate the treatment success rate.@*RESULTS@#The operation time of cold flowable gutta-percha group was significantly lower than that of the control group, which were 51 s and 74 s (P < 0.05); The percentages of pain and discomfort in the two groups were 26.67% and 40.00%, respectively. There were two cases of underfilling and no overfilling in both groups. The percentages of proper filling and tight three-dimensional obturation in the experimental and control groups were 71.11% and 60.00% respectively; and the percentages of tight three-dimensional obturation in the apical third areas were 86.67% and 66.67%, the difference was significant (P < 0.05). There was no significant difference in the three-dimensional tightness between the two groups in the middle and coronal third areas. The percentages of tight three-dimensional obturation in classification Ⅰ groups were 86.67%, 83.33%, 93.33% and 76.67%, 90.00%, 96.67% in experimental and control group, respectively; The percentages of classification Ⅱ/Ⅲ/Ⅳ groups were 86.67%, 86.67%, 100.00% and 46.67%, 86.67%, 100.00%, respectively, and the difference was significant (P < 0.05). There were no apical lesions that occurred in either group during the one-year review period.@*CONCLUSION@#The application of cold flowable gutta-percha on young permanent teeth root canal obturation after apexification can achieve good obturation effect. The root obturation effect in the apical third area is significantly better than that of warm gutta-percha obturation techniques. Cold flowable gutta-percha can shorten the clinical treatment time and ameliorate the patients' therapy comfort.
Subject(s)
Humans , Apexification , Dental Pulp Cavity , Gutta-Percha , Radiography , Root Canal Filling Materials , Root Canal Obturation , Root Canal PreparationABSTRACT
Objective:To investigate the effect of body axis rotation combined with rhythmic breathing training on lower limb motor function and balance ability of stroke patients with hemiplegia. Methods:From June, 2018 to June, 2020, 50 patients with hemiplegia after stroke from Physical Therapy Department of Rehabilitation Hospital Affiliated to Fujian University of Traditional Chinese Medicine were divided into control group (n = 25) and experimental group (n = 25). Both groups accepted routine rehabilitation, while the experimental group accepted body axis rotation and rhythmic breathing training for core muscle strength in addition, for six weeks. The total training time was the same for both groups. They were assessed with Fugl-Meyer Assessment-Lower Extremities (FMA-LE), Berg Balance Scale (BBS), Timed 'Up & Go' Test (TUGT) and 10-meter walking speed (10MWS) before and after treatment. Results:The scores of FMA-LE and BBS, and 10MWS increased in both groups after treatment (|Z| > 4.375, P < 0.001), and increased more in the experimental group than in the control group (|Z| > 3.415, P < 0.01); while the time of TUGT decreased in both groups (Z = -4.372, P < 0.001), and decreased more in the experimental group than in the control group (Z = -2.804, P < 0.01). Conclusion:Body axis rotation combined with rhythmic breathing training for core muscle strength may promote the recovery of lower limb motor function and balance ability for stroke patients with hemiplegia.
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OBJECTIVE@#To evaluate the clinical success of the treatment of maxillary anterior primary incisors caries with composite resin strip crowns.@*METHODS@#Children who presented with severe early childhood caries and were treated comprehensive caries under general anesthesia from January to December 2016 were enrolled in this study. Composite crowns using preformed celluloid crown (3M ESPE, USA) called as "strip crown technique" were applied to treat vital primary incisors with two or multiple surface cavities. Selective etchings of enamel, Universal Bond adhesive and 3M Z350 resin were used to make strip crowns. The patients returned at the end of 6, 12 and 18 months and received clinical and radiologic examinations. Dentists who did not attend the treatment evaluated the strip crowns clinically by modified United States Public Health Service (USPHS) criteria.@*RESULTS@#Four hundred eighteen restorations, placed in 127 children aged 1.17-5.75 years (average of 3.22), were evaluated. The overall retention rate was 97.8% at the end of 6 months, 93.6% at 12 months and 89.2% at 18 months. After 18 months, 28 restorations (6.7%) were totally lost and 4.1% were rated as having lost some resin material. Only four teeth (1.0%) had secondary caries at the end of 18 months and 1.4% teeth had pulpal pathology requiring root canal treatment. Composite crowns had good performance in contour and adjacent contact and improved aesthetics significantly. Twenty-nine teeth (6.9%) showed mild gingivitis and 93.1% showed healthy gingival. 11.2% of the teeth demonstrated color change because plaque accumulation and the polish could remove the stains. The complete loss of strip crowns was mainly related to eating bites.@*CONCLUSION@#Strip crowns performed well for restoring primary incisors with large or multisurface caries for periods of over 18 months. The strip crowns can be a durable and aesthetic restoration for vital carious primary incisors with adequate tooth structure after caries removal. Functional movement is an important cause of complete loss. Because of the high technical sensitivity and its requirement of the cooperation of children, strip crowns are more suitable for older and cooperative children as well as children receiving dental treatment under sedation or general anesthesia.
Subject(s)
Child , Child, Preschool , Humans , Infant , Composite Resins , Crowns , Incisor , Maxilla , Tooth, Deciduous , United StatesABSTRACT
OBJECTIVE@#To evaluate the clinical effect of fluoride releasing flowable resin used in treatment of early enamel caries of children compared with conventional sealant.@*METHODS@#Seventy-six patients, including fifty-two couples of permanent first molars and thirty couples of premolars were selected for this trial. Both sides of all the molars and premolars were diagnosed as early enamel caries based on International Caries Detection and Assessment System (ICDAS) dental caries diagnostic criteria. Using the contralateral control, the teeth were randomly divided into two groups. Molars/premolars in the experimental group were sealed with a fluoride releasing flowable resin; the contralateral molars/premolars were sealed with a conventional fissure sealant as a control group. The retention rate and progress of caries were evaluated at the end of 3, 6, 12 and 24 months.@*RESULTS@#There was no fissure sealant loss or caries progression in both groups 3 and 6 months after sealing the premolars by fluoride releasing flowable resin or conventional fissure sealants. At the end of 12 and 24 months, only one case of fissure sealant loss was observed in conventional fissure sealant group. There was no statistical significance between the two premolar groups. In the first molar group, fluoride releasing flowable resin showed 100%, 98.08%, 90.38% and 88.46% complete retention at the end of 3, 6, 12 and 24 months, respectively. The conventional fissure sealant retention rates were 96.15%, 92.31%, 76.92% and 73.08% at the corresponding time points. The data at the end of 24 months showed that fluoride releasing flowable resin had significantly higher retention rate than the conventional fissure sealant group (P<0.05). The incidence of caries progression at the end of 6, 12 and 24 months were 1.92%, 5.77% and 7.69%, respectively, in the fluoride releasing flowable resin group. In the conventional fissure sealant group, the incidence was 5.77%, 19.23% and 25.00%, respectively. At the end of 12 and 24 months, more significant decrease of caries progress incidence was observed in the fluoride releasing flowable resin group than in the control group, and there was statistical difference between the two groups (P<0.05).@*CONCLUSION@#Compared with conventional fissure sealant, using fluoride releasing flowable resin as a fissure sealant in children enamel caries of permanent molars can improve the sealant preservation rate and effectively prevent enamel caries progress.
Subject(s)
Child , Humans , Dental Caries/prevention & control , Dental Enamel , Fluorides , Molar , Pit and Fissure SealantsABSTRACT
<p><b>OBJECTIVE</b>To identify the functions of the proteins containing the GGDEF or EAL domain in Lactobacillus acidophilus for investigation of the regulatory mechanism of c-di-GMP in this strain.</p><p><b>METHODS</b>The DNA fragments of NH13_07045-GGDEF, NH13_07050 and NH13_07055 from Lactobacillus acidophilus ATCC4356 were amplified by PCR and cloned into the expression vector pMAL-His-c2. After sequencing, the recombinant plasmids were transformed into competent Escherichia coli cells, which were induced by IPTG to express the recombinant proteins fused with maltose binding protein (MBP). The fusion proteins were purified using amylose resin column for diguanylate cyclase (DGC) or phosphodiesterase (PDE) activity assays in vitro followed by analysis with high-performance liquid chromatography (HPLC).</p><p><b>RESULTS</b>The target DNA fragments were obtained by PCR, and their sequences were all identical to that in GenBank. The purified and concentrated fusion proteins, which were identified by SDS-PAGE and Western blotting, had relative molecular masses of 59 kD, 67 kD and 72 kD. HPLC analysis showed no DGC activity in NH13_07045-GGDEF, while PDE activity was found in NH13_07050 but not in NH13_07055.</p><p><b>CONCLUSION</b>We obtained the protein encoded by NH13_07050 that possesses PDE activity in vitro. This protein may facilitate the evaluation of the regulatory function of c-di-GMP in Lactobacillus acidophilus.</p>
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<p><b>OBJECTIVE</b>To establish a sensitive and direct method for detecting the activation of protein kinase C (PKC) using fluorescence resonance energy transfer (FRET) technique.</p><p><b>METHODS</b>HEK293 cells were transfected with C kinase activity reporter (CKAR) plasmid or/and parathyroid receptor 1 plasmid , and after incubation for 72 h, the fluorescence resonance energy transfer was measured with or without parathyroid or TPA stimulation.</p><p><b>RESULTS</b>TPA reduced the efficiency of FRET and increased the emission ratio of CFP/YFP (C/Y) in HEK293 cells transfected with CKAR. PTH(1-34) could increase the emission ratio of C/Y in HEK293 cells co-transfected with CKAR and PTHR1 but not in cells transfected with CKAR.</p><p><b>CONCLUSION</b>FRET analysis using CKAR can be utilized to detect the activation of PKC, which provides a useful means for studying the signaling pathways associated with PKC.</p>