Objective:To investigate the effects of artificial light at night on the growth of mandibles in mice and its regulatory mechanisms.Methods:A mouse model of artificial light at night (night light pollution group) and normal lighting (normal light group) was established by controlling light exposure time, with 4 mice in each group. Micro-CT was employed to analyze the differences in bone quantities of the mandibles between the two groups. Real-time fluorescence quantitative PCR (RT-qPCR) was used to examine the expression levels of osteogenic differentiation and metabolism-related genes in the cortical bone and condylar ossification center of the mandibles. Enzyme-linked immunosorbent assay was utilized to assess the diurnal variation of serum melatonin concentrations between the two groups. The artificial light at night experimental group received daily timed injections of a defined dose of melatonin to restore the diurnal variation of serum melatonin concentration in the mice, while the normal light group and the artificial light at night control group received the same volume of saline. Bone quantities, mandibular tissue morphologies, ossification differentiation in the condylar region and cortical bone, as well as glucose metabolism expression differences were assessed across the three groups.Results:The cortical bone thickness of the mandibles in the artificial light at night group [(0.196±0.017) mm] was significantly less than that in the control group [ (0.228±0.007) mm] ( P=0.029). The bone volume fraction of the condylar ossification center in the artificial light at night group [(36.90±1.09) %] was significantly lower than that in the normal light group [(54.24±1.49) %] ( P<0.001). The length of the mandible in the artificial light at night group [(10.86±0.17) mm] was significantly shorter than that in the normal light group [(11.41±0.32) mm] ( P=0.032). RT-qPCR results indicated that the expressions of osteogenic-related genes alkaline phosphatase, liver/bone/kidney (Alpl), osteocalcin (Ocn), Runt-related transcription factor 2 (Runx2), and osterix (Osx) in the cortical bone and condylar ossification center of mice in the artificial light at night group were significantly lower than those in the normal light group (all P<0.05). The expression of metabolism-related genes protein kinase, AMP activated alpha 1 (Prkaa1), V-type proton ATPase subunit d1 (Atp6v0d1), and cytochrome C oxidase subunit Ⅳ isoform 1 (Cox4i1) in the cortical bone and condylar ossification center of mice in the artificial light at night group were also significantly reduced compared to normal mice (all P<0.01). The serum melatonin concentration peaked 8 hours after lights off in the normal light group, whereas the artificial light at night group exhibited a significantly reduced nocturnal serum melatonin concentration with no apparent peak compared to the normal light group. Micro-CT results demonstrated that after artificial light at night group mice received timed melatonin supplementation daily, the thickness of cortical bone, the bone volume fraction of the condylar ossification center, and the length of the mandible were all significantly higher than those in the artificial light at night group (all P<0.05). Histological staining results indicated that the cortical bone structure of the mandibles in the melatonin supplementation group was more organized than that of the artificial light at night group, with higher brain and muscle ARNT-like 1 (BMAL1) expression ( P=0.003). RT-qPCR results further showed that the expression levels of Prkaa1, Atp6v0d1, and Cox4i1 in the cortical bone and condylar ossification center significantly increased in the melatonin supplementation group compared to the artificial light at night group (all P<0.05), but still significantly lower than those in the normal light group (all P<0.05). Additionally, the RT-qPCR results further revealed that the expression levels of osteogenic differentiation-related genes Alpl, Ocn, and Runx2 in the cortical bone and condylar ossification center of the melatonin supplementation group were significantly higher than those in the artificial light at night group (all P<0.01). Western blotting analysis indicated that the expression levels of glucose metabolism and osteogenic-related proteins RUNX2, OSX, ATP6V0D1, and COX Ⅳ, along with the phosphorylation levels of AMPKα1/α2, were significantly higher in the melatonin supplementation group compared to the artificial light at night group (all P<0.01). Conclusions:Artificial light at night can inhibit melatonin secretion in mice, reduce glucose metabolism in mandibular tissues, and affect both intramembranous and chondrogenic ossification activities, ultimately leading to inadequate mandibular development.

Objective:To investigate the effects of artificial light at night on the growth of mandibles in mice and its regulatory mechanisms.Methods:A mouse model of artificial light at night (night light pollution group) and normal lighting (normal light group) was established by controlling light exposure time, with 4 mice in each group. Micro-CT was employed to analyze the differences in bone quantities of the mandibles between the two groups. Real-time fluorescence quantitative PCR (RT-qPCR) was used to examine the expression levels of osteogenic differentiation and metabolism-related genes in the cortical bone and condylar ossification center of the mandibles. Enzyme-linked immunosorbent assay was utilized to assess the diurnal variation of serum melatonin concentrations between the two groups. The artificial light at night experimental group received daily timed injections of a defined dose of melatonin to restore the diurnal variation of serum melatonin concentration in the mice, while the normal light group and the artificial light at night control group received the same volume of saline. Bone quantities, mandibular tissue morphologies, ossification differentiation in the condylar region and cortical bone, as well as glucose metabolism expression differences were assessed across the three groups.Results:The cortical bone thickness of the mandibles in the artificial light at night group [(0.196±0.017) mm] was significantly less than that in the control group [ (0.228±0.007) mm] ( P=0.029). The bone volume fraction of the condylar ossification center in the artificial light at night group [(36.90±1.09) %] was significantly lower than that in the normal light group [(54.24±1.49) %] ( P<0.001). The length of the mandible in the artificial light at night group [(10.86±0.17) mm] was significantly shorter than that in the normal light group [(11.41±0.32) mm] ( P=0.032). RT-qPCR results indicated that the expressions of osteogenic-related genes alkaline phosphatase, liver/bone/kidney (Alpl), osteocalcin (Ocn), Runt-related transcription factor 2 (Runx2), and osterix (Osx) in the cortical bone and condylar ossification center of mice in the artificial light at night group were significantly lower than those in the normal light group (all P<0.05). The expression of metabolism-related genes protein kinase, AMP activated alpha 1 (Prkaa1), V-type proton ATPase subunit d1 (Atp6v0d1), and cytochrome C oxidase subunit Ⅳ isoform 1 (Cox4i1) in the cortical bone and condylar ossification center of mice in the artificial light at night group were also significantly reduced compared to normal mice (all P<0.01). The serum melatonin concentration peaked 8 hours after lights off in the normal light group, whereas the artificial light at night group exhibited a significantly reduced nocturnal serum melatonin concentration with no apparent peak compared to the normal light group. Micro-CT results demonstrated that after artificial light at night group mice received timed melatonin supplementation daily, the thickness of cortical bone, the bone volume fraction of the condylar ossification center, and the length of the mandible were all significantly higher than those in the artificial light at night group (all P<0.05). Histological staining results indicated that the cortical bone structure of the mandibles in the melatonin supplementation group was more organized than that of the artificial light at night group, with higher brain and muscle ARNT-like 1 (BMAL1) expression ( P=0.003). RT-qPCR results further showed that the expression levels of Prkaa1, Atp6v0d1, and Cox4i1 in the cortical bone and condylar ossification center significantly increased in the melatonin supplementation group compared to the artificial light at night group (all P<0.05), but still significantly lower than those in the normal light group (all P<0.05). Additionally, the RT-qPCR results further revealed that the expression levels of osteogenic differentiation-related genes Alpl, Ocn, and Runx2 in the cortical bone and condylar ossification center of the melatonin supplementation group were significantly higher than those in the artificial light at night group (all P<0.01). Western blotting analysis indicated that the expression levels of glucose metabolism and osteogenic-related proteins RUNX2, OSX, ATP6V0D1, and COX Ⅳ, along with the phosphorylation levels of AMPKα1/α2, were significantly higher in the melatonin supplementation group compared to the artificial light at night group (all P<0.01). Conclusions:Artificial light at night can inhibit melatonin secretion in mice, reduce glucose metabolism in mandibular tissues, and affect both intramembranous and chondrogenic ossification activities, ultimately leading to inadequate mandibular development.

The prevalence of Class III malocclusion varies among different countries and regions. The populations from Southeast Asian countries (Chinese and Malaysian) showed the highest prevalence rate of 15.8%, which can seriously affect oral function, facial appearance, and mental health. As anterior crossbite tends to worsen with growth, early orthodontic treatment can harness growth potential to normalize maxillofacial development or reduce skeletal malformation severity, thereby reducing the difficulty and shortening the treatment cycle of later-stage treatment. This is beneficial for the physical and mental growth of children. Therefore, early orthodontic treatment for Class III malocclusion is particularly important. Determining the optimal timing for early orthodontic treatment requires a comprehensive assessment of clinical manifestations, dental age, and skeletal age, and can lead to better results with less effort. Currently, standardized treatment guidelines for early orthodontic treatment of Class III malocclusion are lacking. This review provides a comprehensive summary of the etiology, clinical manifestations, classification, and early orthodontic techniques for Class III malocclusion, along with systematic discussions on selecting early treatment plans. The purpose of this expert consensus is to standardize clinical practices and improve the treatment outcomes of Class III malocclusion through early orthodontic treatment.
Humans ; Malocclusion, Angle Class III/classification* ; Orthodontics, Corrective/methods* ; Consensus ; Child

Bone regeneration remains a great clinical challenge. Low intensity near-infrared (NIR) light showed strong potential to promote tissue regeneration, offering a promising strategy for bone defect regeneration. However, the effect and underlying mechanism of NIR on bone regeneration remain unclear. We demonstrated that bone regeneration in the rat skull defect model was significantly accelerated with low-intensity NIR stimulation. In vitro studies showed that NIR stimulation could promote the osteoblast differentiation in bone mesenchymal stem cells (BMSCs) and MC3T3-E1 cells, which was associated with increased ubiquitination of the core circadian clock protein Cryptochrome 1 (CRY1) in the nucleus. We found that the reduction of CRY1 induced by NIR light activated the bone morphogenetic protein (BMP) signaling pathways, promoting SMAD1/5/9 phosphorylation and increasing the expression levels of Runx2 and Osterix. NIR light treatment may act through sodium voltage-gated channel Scn4a, which may be a potential responder of NIR light to accelerate bone regeneration. Together, these findings suggest that low-intensity NIR light may promote in situ bone regeneration in a CRY1-dependent manner, providing a novel, efficient and non-invasive strategy to promote bone regeneration for clinical bone defects.
Animals ; Rats ; Bone Morphogenetic Protein 2/metabolism* ; Bone Regeneration ; Cell Differentiation ; Circadian Clocks ; Cryptochromes/metabolism* ; Osteoblasts/metabolism* ; Osteogenesis ; Transcription Factors/metabolism*

Cardiovascular disease is still the leading cause of mortality worldwide. Vascular endothelial dysfunction is viewed as the initial step of most cardiovascular diseases. Many studies have indicated that periodontal pathogens, especially Porphyromonas gingivalis, are closely correlated with vascular endothelial homeostasis, but the function of P. gingivalis and the underlying mechanisms are still elusive. To illuminate the effects and elucidate the mechanisms of P. gingivalis on endothelial structural integrity, we developed P. gingivalis infection models in vivo and in vitro. Endothelial cell proliferation, differentiation and apoptosis were detected. Here, we showed that P. gingivalis can impair endothelial integrity by inhibiting cell proliferation and inducing endothelial mesenchymal transformation and apoptosis of endothelial cells, which reduce the cell levels and cause the endothelium to lose its ability to repair itself. A mechanistic analysis showed that TLR antagonist or NF-κB signalling inhibitor can largely rescue the damaged integrity of the endothelium caused by P. gingivalis, suggesting that TLR-NF-κB signalling plays a vital role in vascular endothelial homeostasis destroyed by P. gingivalis. These results suggest a potential intervention method for the prevention and treatment of cardiovascular disease.

OBJECTIVE: To explore the genotype-phenotype correlation of Cardio-facio-cutaneous syndrome (CFCS) caused by MAP2K1 gene variants. METHODS: Genomic DNA was extracted from peripheral blood sample from a child patient and his parents. Whole exome sequencing (WES) was carried out for the patient. Suspected variant was verified by Sanger sequencing. RESULTS: The patient was a 1-year-8-month old Chinese male who manifested short stature, psychomotor retardation, relative macrocephaly, distinctive facial features, and congenital heart disease. WES test revealed a heterozygous missense c.389A>G (p.Tyr130Cys) variant in the MAP2K1 gene. Sanger sequencing has confirmed the variant as de novo. According to ACMG/AMP guidelines, the variant was classified as pathogenic. CONCLUSION Compared with previously reported CFCS cases due to MAP2K1 variants. The patient showed obvious behavioral problems, good appetite and tricuspid regurgitation, which may to be novel features for CFCS.
China ; Ectodermal Dysplasia ; genetics ; Facies ; Failure to Thrive ; genetics ; Genetic Association Studies ; Genetic Variation ; Heart Defects, Congenital ; genetics ; Heterozygote ; Humans ; Infant ; MAP Kinase Kinase 1 ; genetics ; Male ; Mutation ; Whole Exome Sequencing

Objective To explore the effects of bone marrow mesenchymal stem cells (MSCs) transplantation on receptor-interacting protein kinase 1 (RIP1) and RIP3 in rat brain after cardiac arrest (CA).Methods Sprague Dawley (SD) rats were randomly (random number) divided into sham group (n=8),CA group (n=8) and MSCs group (n=8).Animals were subjected to asphyxial cardiac arrest and followed by cardiopulmonary resuscitation (CPR).In MSCs group or CA group,animals received intravenous injection of 1 × 106 MSCs in 0.5 mL phosphate buffer solution (PBS) or 0.5 mL PBS alone at 1 h after successful resuscitation.Neurological deficit scores (NDS) were assessed at 3 d after CPR.Donor MSCs in brain were detected under a fluorescent microscope.HE staining of brain tissue was performed to observe necrotic neurons.Western blot analysis was performed to measure the levels of RIP1 and RIP3 in brain.Multiple comparisons were made by analysis of variance or Kruskal-Wallis H test.Results At 3 d after CPR,MSCs group demonstrated higher NDS than CA group [72.5(71.5,73.2) vs.63.0(62.5,64.1),Z=3.376,P=0.001].DAPI-labeled MSCs were primarily observed in the cerebral cortex.The percentage of necrotic neurons in MSCs group was significantly lower than that in CA group [(29.6±5.9)％ vs.(57.2±6.4)％,t=8.922,P＜0.01].The levels of RIP1 and RIP3 expression in brain in MSCs group were significantly lower than those in CA group [RIP1:0.227(0.193,0.243) vs.0.599(0.535,0.629),Z=3.151,P=0.001;RIP3:0.217(0.203,0.274) vs.0.543(0.533,0.555),Z=3.361,P=0.001].Conclusion MSCs transplantation improves neurological function after CPR from CA in rats likely associated with inhibiting necroptosis.

Objective To investigate the improvement of ischemic hypoxic injury of brain after the transplantation of bone marrow mesenchymal stem cells (BMSCs).Methods Rats were randomly (random number) divided into sham operation group,cardiac arrest group and BMSCs treatment group (n =10 in each group).The model of cardiac arrest was induced by asphyxia.One hour after restoration of spontaneous circulation (ROSC),green fluorescent protein labeled BMSCs were transplanted via tail vein injection.At 3 and 7 days after transplantation,frozen sections of hippocampus was stained with hematoxylin-eosin (HE).The rest of brain tissue was weighed by an electronic balance.Brain water content (％) was calculated as (wet weight-dry weight) / wet weight × 100％.Results ①BMSCs were observed in hippocampus at 3 and 7 days after transplantation under fluorescent microscopy.②Compared with sham operation group and BMSCs treatment group,brain water content in cardiac arrest group was higher (all P ＜ 0.05).HE staining results showed that BMSCs transplantation could lessen hypoxia ischemia damage on brain.Conclusions BMSCs reduced the neurons damage induced by cardiac arrest and promoted neurological function recovery.

Objective To investigate the distribution of alpha2-HS glycoprotein (AHSG) gene polymorphisms and the relationships of AHSG gene polymorphisms with atherosclerosis as well as serum bone related biochemical mark-era. Methods Blood samples of 344 hospitalized female patients, aged 20 ～ 80 years, were sampled for serum bone alkaline phosphatase, cross-linked N-telopeptide of collagen type Ⅰ, cross-linked C-telopeptide of collagen type Ⅰ , osteoprotegrin and leptin were determined by ELISA. Serum TC,TG and calcium content were detected. Poly-morphism of AHSG gene was detected by polymerase chain reaction fragment length polymorphisms (PCR-RFLP) of restriction enzyme Sac Ⅰ. BMD (Norland XR-36) of the anteroposterior spine (AP), supine lateral spine (Lat) and femoral neck (FN) were measured. Morphological changes in the aorta and bone of type GG patient were detected by pathological microscopy. IMT were measured by color doppler ultrasound equipment(SEQUOIAS12). Results (1) The genotype frequency of CC, CG, and GG were 59.7%, 25.1% and 15.2% respectively in all elderly female patients. There were significant difference in blood lipids, Ca~(2+) and serum bone relative biochemical markers to different AHSG genotypes. (2)There were significant differences in the BMD of the AP, Lat, FN and IMT and the serum biochemical markers among the CC, CG and GG genotypes. (3)GG-female patients bone tissue pathology section verify the AHSG polymorphism genetic mutation and atherosclerosis, osteoporosis development of the relationship. Conclusion There was close relationship among AHSG polymorphism variation and the incidence of arteriosclerosis and osteoporosis in elderly female.