Role of oncogenic long noncoding RNA KCNQ1OT1 in colon cancer

The role of lncRNA KCNQ1 opposite strand/antisense transcript 1(KCNQ1OT1)in colon cancer involves various tumorigenic processes and has been studed widely.However,the mechanism by which it promotes colon cancer remains unclear.Retrovirnl vector pSEB61 was retroftted in established HCT116 siKCN and SW480-siKCN cells to silence KCNQ1 OT1.Cellular proliferation was measured using CCK8 assay,and flow cytometry(FCM)detected cell cydle changes.RNA sequencing(RNA Seq)analysis showed differentially expressed genes(DEGs).Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were carried out to analyze enriched functions and signaling pathways.RT-qPCR,immunofluorescence,and western blotting were carried out to validate downstream gene expressions.The effects of tumorigenesis were evaluated in BALB/c nude mice by tumor xenografts.Our data revealed that the silencing of KONQ1OT1 in HCT116 and SW480 cells slowed cell growth and decreased the number of cells in the G2/M phase.RNA-Seq analysis showed the data of DEGs enriched in various GO and KEGG pathways such as DNA replication and cell cyde.RT qPCR,immunofluorescence,and western blotting confirmed downstream CCNE2 and PCNA gene expressions.HCT116 siKCN cells signifcantly suppressed tumorigenesis in BALB/c nude mice.Our study suggests that lncRNA KCNQ1OT1 may provide a promising therapeutic strategy for colon cancer.

Comparison of Success Rate and Radiological Bone Loss in Patients with Mini-Implant versus Conventional-Sized Implant Removable Prostheses: A Meta-Analysis

Background: The objective of this meta-analysis was to assess whether mini-implants have added benefit in terms of implants success rate and average bone loss over conventional-sized implants after one year of follow-up. Methods: An electronic search of randomized clinical trials was conducted in MEDLINE (via PubMed), Cochrane Central Register of Clinical Trials (CENTRAL) and Web of Science for studies including complete or partial edentulous patients requiring two or four mini-implants or conventional/ standard-sized implants in the maxilla or mandible for implant-supported removable prostheses who completed 12 months of follow-up. Results: The search provided 194 unique articles which were screened for title and abstract. Screening generated 12 articles which went through full-text analysis using eligibility criteria, and 4 articles were included for meta-analysis. Meta-analysis of these studies indicated a non-significant difference in the success rate between the two interventions (OR = 1.69 [0.74, 3.85;p = 0.21]). Bone loss estimates resulted in a significant bone reduction (Mean Difference = -0.74 [-0.95, -0.53;p < 0.05]) in favor of two mini-implants when compared with two conventional-sized implants, but when compared four mini- with two conventional-sized implants, the estimates were non-significant (Mean Difference = -0.24 [-0.69, 0.20;p = 0.29]). Conclusion: The current evidence does not provide solid evidence of the benefits of one intervention over the other. More studies with follow-up times of 10 and more years are needed as current studies have described the short-term outcomes.

Synthesis and potential osteogenic applications of Wnt3a-loaded ZIF-8 nanoparticles

The Wnt signaling pathway plays a critical role in bone homeostasis,and the related protein therapy strategies have been reported to have great potential in osseointegration;however,they face formidable challenges such as complex external environments and unavoidable protein denaturation.In this work,we report a novel approach combining the synthesis of metal–organic frameworks(MOFs)and protein encapsulation in a one-pot process based on zeolitic imidazolate framework-8(ZIF-8)and Wnt3a protein,with improved biomechanical behavior and enhanced protein biological response.This combination was designed to enhance the Wnt3a protein function through the improved chemical stability provided by the ZIF-8 crystals.Additionally,the zinc ions contained in the ZIF-8 crystals induced bone homeostasis,further favoring the osteogenesis.The results showed that the Wnt3a protein-loaded ZIF-8 crystals served as efficient drug delivery vehicles to promote osteogenesis,preventing protein denaturation.In particular,Wnt3a-loaded ZIF-8 nanoparticles(Wnt3a@ZIF-8 NPs)had higher efficacy on bone marrow mesenchymal stem cells(BMSCs)than ZIF-8 NPs or Wnt3a proteins,contributing to the osteogenesis through ZIF-8 crystals and intracellular Wnt3a proteins released from Wnt3a@ZIF-8 NPs.Furthermore,polymerase chain reaction(PCR)analysis showed that the osteogenic pathways were upregulated.Overall,the present one-pot process can open up new avenues to develop signaling protein-delivery systems for applications in protein therapy strategies.

Modified orthodontic treatment of substitution of canines by first premolars:A case report

BACKGROUND Canines are the most important teeth in the dentition.Usually,doctors choose to remove premolars rather than canines.Canine extraction is extremely rare in orthodontic treatment.However,dentists sometimes encounter situations in which canines require extraction due to defects caused by improper medical treatment.CASE SUMMARY The present study reports a case of a class II adult patient treated with the extraction of maxillary canines and right mandibular second premolar.After postactive treatment for 28 mo,then the canines were substituted by the upper first premolar,a satisfactory occlusal was established,the lips were competent,and the profile was improved.Intraoral pictures and X-ray data retrieved 3 years after the end of orthodontic treatment demonstrated the possibility of canine extraction and premolar substitution of canines in function and beauty.CONCLUSION The extraction of canines and substitution by first premolars could be a feasible orthodontic treatment.

mTORC1 signaling pathway regulates tooth repair

Tooth germ injury can lead to abnormal tooth development and even tooth loss,affecting various aspects of the stomatognathic system including form,function,and appearance.However,the research about tooth germ injury model on cellular and molecule mechanism of tooth germ repair is still very limited.Therefore,it is of great importance for the prevention and treatment of tooth germ injury to study the important mechanism of tooth germ repair by a tooth germ injury model.Here,we constructed a Tg(dlx2b:Dendra2-NTR)transgenic line that labeled tooth germ specifically.Taking advantage of the NTR/Mtz system,the dlx2b+tooth germ cells were depleted by Mtz effectively.The process of tooth germ repair was evaluated by antibody staining,in situ hybridization,Ed U staining and alizarin red staining.The severely injured tooth germ was repaired in several days after Mtz treatment was stopped.In the early stage of tooth germ repair,the expression of phosphorylated 4E-BP1 was increased,indicating that mTORC1 is activated.Inhibition of mTORC1 signaling in vitro or knockdown of mTORC1 signaling in vivo could inhibit the repair of injured tooth germ.Normally,mouse incisors were repaired after damage,but inhibition/promotion of mTORC1 signaling inhibited/promoted this repair progress.Overall,we are the first to construct a stable and repeatable repair model of severe tooth germ injury,and our results reveal that mTORC1 signaling plays a crucial role during tooth germ repair,providing a potential target for clinical treatment of tooth germ injury.

The RNA-binding protein Musashi2 governs osteoblast-adipocyte lineage commitment by suppressing PPARγsignaling

Osteoporosis caused by aging is characterized by reduced bone mass and accumulated adipocytes in the bone marrow cavity. How the balance between osteoblastogenesis and adipogenesis from bone marrow mesenchymal stem cells(BMSCs) is lost upon aging is still unclear. Here, we found that the RNA-binding protein Musashi2(Msi2) regulates BMSC lineage commitment. Msi2 is commonly enriched in stem cells and tumor cells. We found that its expression was downregulated during adipogenic differentiation and upregulated during osteogenic differentiation of BMSCs. Msi2 knockout mice exhibited decreased bone mass with substantial accumulation of marrow adipocytes, similar to aging-induced osteoporosis. Depletion of Msi2 in BMSCs led to increased adipocyte commitment. Transcriptional profiling analysis revealed that Msi2 deficiency led to increased PPARγ signaling.RNA-interacting protein immunoprecipitation assays demonstrated that Msi2 could inhibit the translation of the key adipogenic factor Cebpα, thereby inhibiting PPAR signaling. Furthermore, the expression of Msi2 decreased significantly during the aging process of mice, indicating that decreased Msi2 function during aging contributes to abnormal accumulation of adipocytes in bone marrow and osteoporosis. Thus, our results provide a putative biochemical mechanism for aging-related osteoporosis, suggesting that modulating Msi2 function may benefit the treatment of bone aging.

Expert consensus on early childhood caries management

Early childhood caries(ECC)is a significant chronic disease of childhood and a rising public health burden worldwide.ECC may cause a higher risk of new caries lesions in both primary and permanent dentition,affecting lifelong oral health.The occurrence of ECC has been closely related to the core microbiome change in the oral cavity,which may be influenced by diet habits,oral health management,fluoride use,and dental manipulations.So,it is essential to improve parental oral health and awareness of health care,to establish a dental home at the early stage of childhood,and make an individualized caries management plan.Dental interventions according to the minimally invasive concept should be carried out to treat dental caries.This expert consensus mainly discusses the etiology of ECC,caries-risk assessment of children,prevention and treatment plan of ECC,aiming to achieve lifelong oral health.

SATB2:A versatile transcriptional regulator of craniofacial and skeleton development,neurogenesis and tumorigenesis,and its applications in regenerative medicine

SATB2(special AT-rich sequence-binding protein 2)is a member of the special AT-rich binding protein family.As a transcription regulator,SATB2 mainly integrates higher-order chromatin organization.SATB2 expression appears to be tissue-and stage-specific,and is governed by several cellular signaling molecules and mediators.Expressed in branchial arches and osteoblast-lineage cells,SATB2 plays a significant role in craniofacial pattern and skeleton development.In addition to regulating osteogenic differentiation,SATB2 also displays versatile functions in neural development and cancer progression.As an osteoinductive factor,SATB2 holds great promise in improving bone regeneration toward bone defect repair.In this review,we have summarized our current understanding of the physiological and pathological functions of SATB2 in craniofacial and skeleton development,neurogenesis,tumorigenesis and regenerative medicine.

Liquiritigenin promotes osteogenic differentiation and prevents bone loss via inducing auto-lysosomal degradation and inhibiting apoptosis

Osteoporosis(OP)is a debilitating skeletal abnormality involving bone remodeling and bone cell homeostasis characterized by decreased bone strength and high fracture risk.A novel therapeutic intervention for OP by manipulating cellular autophagy-apoptosis processes to promote skeletal homeostasis is presented.Protective effects of the naturally occurring plant extract Liquiritigenin(LG)were demonstrated in an ovariectomy(OvX)-OP mouse model and preosteoblast MC3T3-E1 cells.Micro-CT and histological staining assessments of skeletal phenotype were applied alongside detection of autophagy activity in osteocytes and MC3T3-E1 cells by transmission electron microscopy(TEM).The effects of LG on chloroquine(CQ)-and the apoptosis-inducing TS-treated osteogenic differentiations and status of lysosomes within MC3T3-E1 cells were analyzed by Neutral red,Alizarin red S and alkaline phosphatase(ALP)staining and Western blot assays.Treatment with LG prevented bone loss,increased osteogenic differentiation in vivo and in vitro,and inhibited osteoclast formation to some extent.TEM analyses revealed that LG can improve auto-lysosomal degradation within osteocytes from OVX mice and MC3T3-E1 cells.The abnormal status of lysosomes associated with CQ and TS treatments was notably alleviated by LG which also reduced levels of apoptosis-induced inhibition of osteogenic differentiation and averted abnormal osteogenic differentiation as a consequence of a blockage in autolysosome degradation.Overall,LG stimulates bone growth in Oovx mice through increased osteogenic differentiation and regulation of autophagyapoptosis mechanisms,presenting an auspicious natural therapy for Op.

Effect of 3D anchorage attachment on the alleviating tipping/extrusion of premolars for en-mass distalization of maxillary molars with clear aligners:A finite element study

This study aimed to explore the optimal invisible orthodontic force system during the en-mass distalization of two maxillary molars to minimize the side effect of anchorage loss by changing the direction of the application of the orthodontic force system.A high bio-fidelity 3D finite element model including maxilla,periodontal ligament,dentition,clear aligner,3D anchorage attachment and mini-implant was established.Different lengths of lateral hooks of 3D-printed anchorage attachments and mini-implant positions into the palatal alveolus were considered.A 200 g distal force was applied to the lateral hooks of different horizontal lengths(3.26 mm,6.52 mm and 9.78 mm)with the mini-implant as the application point.Using ABAQUS software,orthodontic tooth movements under 12 different clinical treatment designs were analyzed and calculated.The 3D anchorage attachment enhanced the anchorage of anterior teeth and alleviated the tipping/extrusion of premolars.In contrast to without clear aligners,length of the lateral hook had a negligible effect on both mesial tipping and buccal tipping with clear aligners,which could then be ignored.The change in mesial tipping was less and nearly remained constant despite of the different heights of the mini-implant.The 3D anchorage attachment assisted clear aligner can avoid the side effects of anterior tooth proclination caused by insufficient anchorage.The length of the lateral hook,and height of the mini-implant in this invisible orthodontic force system hardly affects the tooth movement of anchorage units.Clear aligners can effectively control the rotation and tipping of anchorage units caused by 3D anchorage attachment.

pH and lipase-responsive nanocarrier-mediated dual drug delivery system to treat periodontitis in diabetic rats

Precise and controlled drug delivery to treat periodontitis in patients with diabetes remains a significant clinical challenge.Nanoparticle-based drug delivery systems offer a potential therapeutic strategy;however,the low loading efficiency,non-responsiveness,and single effect of conventional nanoparticles hinder their clinical application.In this study,we designed a novel self-assembled,dual responsive,and dual drug-loading nanocarrier system,which comprised two parts:the hydrophobic lipid core formed by 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-Poly(ethylene glycol)(DSPE-PEG)loaded with alpha-lipoic acid(ALA);and a hydrophilic shell comprising a poly(amidoamine)dendrimer(PAMAM)that electrostatically adsorbed minocycline hydrochloride(Mino).This unique design allows the controlled release of antioxidant/ALA under lipase stimulation from periodontal pathogens and antimicrobial/Mino under the low pH of the inflammatory microenvironment.In vivo and in vitro studies confirmed that this dual nanocarrier could inhibit the formation of subgingival microbial colonies while promoting osteogenic differentiation of cells under diabetic pathological conditions,and ameliorated periodontal bone resorption.This effective and versatile drug-delivery strategy has good potential applications to inhibit diabetes-associated periodontal bone loss.

Bioinspired Andrias davidianus-Derived wound dressings for localized drug-elution

Local drug delivery has received increasing attention in recent years.However,the therapeutic efficacy of local delivery of drugs is still limited under certain scenarios,such as in the oral cavity or in wound beds after resection of tumors.In this study,we introduce a bioinspired adhesive hydrogel derived from the skin secretions of Andrias davidianus(SSAD)as a wound dressing for localized drug elution.The hydrogel was loaded with aminoguanidine or doxorubicin,and its controlled drug release and healing-promoting properties were verified in a diabetic rat palatal mucosal defect model and a C57BL/6 mouse melanoma-bearing model,respectively.The results showed that SSAD hydrogels with different pore sizes could release drugs in a controllable manner and accelerate wound healing.Transcriptome analyses of the palatal mucosa suggested that SSAD could significantly upregulate pathways linked to cell adhesion and extracellular matrix deposition and had the ability to recruit keratinocyte stem cells to defect sites.Taken together,these findings indicate that property-controllable SSAD hydrogels could be a promising biofunctional wound dressing for local drug delivery and promotion of wound healing.

A custom-designed panel sequencing study in 201 Chinese patients with craniosynostosis revealed novel variants and distinct mutation spectra

Craniosynostosis is a rare disease in which one or more of the cranial sutures in an infant skull prematurely fuses by turning into bone,with a prevalence of 1 in 2,000—2,500 individuals from reports in Western countries(Wilkie et al.,2017).It may restrict the growth of the brain,leading to some degree of morphological and functional abnormalities,and may affect the neurocognitive function of infants(Lattanzi et al.,2017).Genetic variants underlying craniosynostosis have been identified in cohort studies in Western populations.

Bioinspired drug-delivery system emulating the natural bone healing cascade for diabetic periodontal bone regeneration

Diabetes mellitus(DM)aggravates periodontitis,resulting in accelerated periodontal bone resorption.Disordered glucose metabolism in DM causes reactive oxygen species(ROS)overproduction resulting in compromised bone healing,which makes diabetic periodontal bone regeneration a major challenge.Inspired by the natural bone healing cascade,a mesoporous silica nanoparticle(MSN)-incorporated PDLLA(poly(DL-lactide))-PEG-PDLLA(PPP)thermosensitive hydrogel with stepwise cargo release is designed to emulate the mesenchymal stem cell“recruitment-osteogenesis”cascade for diabetic periodontal bone regeneration.During therapy,SDF-1 quickly escapes from the hydrogel due to diffusion for early rat bone marrow stem cell(rBMSC)recruitment.Simulta-neously,slow degradation of the hydrogel starts to gradually expose the MSNs for sustained release of metformin,which can scavenge the overproduced ROS under high glucose conditions to reverse the inhibited osteogenesis of rBMSCs by reactivating the AMPK/β-catenin pathway,resulting in regulation of the diabetic microenvironment and facilitation of osteogenesis.In vitro experiments indicate that the hydrogel markedly restores the inhibited migration and osteogenic capacities of rBMSCs under high glucose conditions.In vivo results suggest that it can effectively recruit rBMSCs to the periodontal defect and significantly promote periodontal bone regeneration under type 2 DM.In conclusion,our work provides a novel therapeutic strategy of a bioinspired drug-delivery system emulating the natural bone healing cascade for diabetic periodontal bone regeneration.

Metformin ameliorates HMGB1-mediated oxidative stress through mTOR pathway in experimental periodontitis

Periodontitis is an oral chronic inflammatory disease.Inhibiting tissue destruction and promoting tissue regeneration are important means for the treatment of periodontitis.Metformin not only has hypoglycemic effect but also has anti-inflammatory effect.Metformin has been shown to inhibit oxidative stress and activate autophagy through AMPK/mTOR pathway.High mobility group box 1(HMGB1)has been implicated in the pathogenesis of many inflammatory diseases including periodontitis,it can participate in the induction of oxidative stress.HMGB1 is an autophagy regulator under oxidative stress,which can activate mTOR pathway.However,it is not clear whether metformin is related to HMGB1 and its mechanism in the process of periodontitis.Cell viability and expression of inflammatory cytokines were clarified by Cell Counting Kit-8,real-time PCR and enzyme-linked immunosorbent assay.Western blot and immunofluorescence were conducted to determine HMGB1 intracellular localization and expression of autophagy-associated proteins in vitro.Experimental periodontitis mice model was induced by administering a ligature.Immunohistochemistry was performed to detect the expression and localization of HMGB1 in vivo.The results of CCK-8,real-time PCR,enzyme-linked immunosorbent assay,Western blot and immunofluorescence showed lipopolysaccharide(LPS)treatment inhibited cell viability,and increased HMGB1 expression at a dose-independent manner.Metformin can reduce the effect of LPS.It also improves autophagy pathway inhibited by LPS and down-regulates mTOR expression.In addition,metformin attenuated alveolar bone resorption induced by ligation.This study provides new evidence for that metformin is a potential drug for the treatment of periodontitis and HMGB1 may be a potential target for periodontal intervention.

Efficient scarless skin regeneration enabled by loading micronized amnion in a bioinspired adhesive wound dressing

Complete skin reconstruction is a hierarchical,physiological assembly process involving healing of the epidermis,dermis,vasculature,nerves,and cutaneous appendages.To date,few works have reported complete skin regeneration,particularly lacking vascular structures and hair follicles after full skin defects.In this study,a hydrogel derived from the skin secretion of Andrias davidianus(SSAD)that features adhesiveness was used as a bioactive scaffold to load micronized amnion(MA).The SSAD hydrogel was found to promote the migration and proliferation of amnion stem cells and human keratinocytes,as well as inhibit their apoptosis in vitro.In a rat full-skin defect model,the regeneration of skin appendages was observed at the wound area,achieving scarless healing.Transcriptome analyses further validated that SSAD could positively regulate cell migration,proliferation,and differentiation.These functions might be attributed to the abundant growth factors present in the SSAD.Synergized by the delivery of MA,SSAD loaded with the MA could achieve a significantly better skin regeneration effect than SSAD or MA used alone,providing a simple yet highly effective means to obtain complete,scarless skin regeneration,suggesting favorable potential for clinical translation.

Beclin-1/LC3-II dependent macroautophagy was uninfluenced in ischemia-challenged vascular endothelial cells

Autophagy has been extensively studied and occurs in many biological settings.However,a question remains as to whether ischemia enhances Beclin-1/LC3-II-dependent macroautophagy in vascular endothelial cells,as has been previously thought.Furthermore,the effect of the level of autophagy on cell or skin flap survival still requires elucidation.We created a lethal ischemia model in human umbilical vascular endothelial cells(HUVECs),performed quantitative proteomics and bioinformatics analyses,and verified the autophagic status and effect both in vitro and in vivo.The significantly upregulated proteins encoded by autophagy-related genes(ATGs)included ATG2A,ATG3,ATG4B,ATG5,ATG7,ATG9A,ATG12,ATG16,and ATG101.The significantly enhanced lysosomal proteins were cathepsin B,cathepsin D,lysosome-associated membrane protein 1(LAMP1),and LAMP2.However,the differentially expressed proteins excluded Beclin-1,microtubule-associated protein light chain 3(LC3)-I,and LC3-II.Western blot analyses verified that the protein expression levels of Beclin-1,LC3-I,and LC3-II were neither upregulated nor downregulated in ischemia-challenged HUVECs.The autophagic status was not enhanced by rapamycin in ischemic HUVECs but appeared to be inhibited by chloroquine.Our in vivo study on rats showed that a downregulation in autophagic status jeopardized skin flap survival.In conclusion,Ischemia neither enhanced nor inhibited Beclin-1/LC3-II-dependent canonical macroautophagy both in vitro and in vivo,in contradiction to previous studies.An appropriate autophagic homeostasis can minimize cell or skin flap damage.