Objective To study the three-dimensional changes of soft and hard tissue in male patients with unilateral non-syndromic cleft lip and palate in the mixed dentition period before and after maxillary protraction.Methods Twenty male patients with unilateral non-syndromic cleft lip and palate in the mixed dentition period treated by maxillary anterior traction in the Department of Orthodontics of Affiliated Stomatological Hospital of Nanjing Medical University were selected(average age(10.6±1.23)years old).Cone beam CT was taken before and after treatment.Dolphin 3D 11.95 software was used for three-dimensional measurement and analysis.SPSS 25.0 software package was used for statistical analysis.The self-controlled paired t test was used to compare the changes in soft and hard tis-sues of male patients with unilateral cleft lip and palate before and after treatment.The changes in the anterior displacement of the ANS point,the anterior displacement of point A,and the posterior displacement of point B were tested using the one-sample t test.Results The sagittal skeletal changes were significantly increased in ∠ SNA(P＜0.01),∠ANB(P＜0.01),Y axis(P＜0.05),the forward displacement of ANS point(P＜0.01)and A point(P＜0.01)and the backward displacement of B point(P＜0.01),but ∠SNB(P＜0.05)was decreased significantly.The vertical skeletal changes showed that ∠MP-FH(P＜0.01),∠MP-SN(P＜0.05)and the dis-tance of ANS-Me(P＜0.05)were increased significantly,but ∠SN-PP(P＜0.01)was decreased significantly.The dental changes inclu-ding ∠U1-NA(P＜0.01),the distance of U1-NA(P＜0.01),∠U1-SN(P＜0.01),overjet and the Wits were increased significantly,but ∠ L1-NB(P＜0.01),the distance of L1-NB(P＜0.01)and ∠L1-MP(P＜0.01)were decreased significantly.The changes of soft tis-sue including ∠ S-Ns-Sn(P＜0.01),∠ Sn-Ns-Bs(P＜0.01),the distance of UL-EP(P＜0.01)and LL-UL(P＜0.01)were increased sig-nificantly.Conclusion After the treatment of maxillary protraction,the forward growth of maxilla will be possibly promoted on patients with cleft lip and palate in the peak of growth timing,as well as the intermaxillary relationship and soft tissue profile,but the side effects should be paid attention to.

Objective:To systematically investigate the causal effects of exposure factors on nonsyndromic cleft lip with or without cleft palate (NSCL/P) using a phenome-wide mendelian randomization (MR-PheWAS) framework and identify pleiotropic loci.Methods:This study integrated genome-wide association study (GWAS) data for NSCL/P, including 1 069 cases and 1 724 controls, and systematically evaluated causal associations between exposures and NSCL/P using the MR-PheWAS framework. GWAS summary data for 2 106 Asian population-exposure phenotypes were obtained from the IEU OpenGWAS database. The inverse-variance weighted (IVW) method served as the core causal inference model, supplemented by weighted median and MR-Egger regression to verify the robustness of causal associations. Additionally, multivariable MR analysis was conducted to adjust for confounding effects, alongside sensitivity tests (Cochran′s Q and MR-PRESSO). Genetic correlations were analyzed using LD Score regression, and cross-phenotype pleiotropy analysis (PLACO/CPASSOC) was employed to identify shared genetic loci. Pathway enrichment and gene annotation data were integrated to explore potential biological mechanisms.Results:MR analysis identified serum calcium ( OR=0.12, P=0.019), high-density lipoprotein (HDL, OR=0.61, P=0.039), and mean corpuscular hemoglobin concentration (MCHC, OR=0.39, P=0.032) as protective factors, whereas serum sodium ( OR=21.41, P=0.013) was a risk factor. Furthermore, in subsequent analyses of genetic correlation and genetic overlap, a strong association was observed between serum calcium and NSCL/P. Cross-trait analysis localized pleiotropic loci to 16q24.2 and 3q21.1, involving CASR and CSTA, with significant enrichment in vitamin D response pathways. Conclusions:Numerous environmental exposure factors may have a causal impact on the outcomes of NSCL/P, and metabolic homeostasis (especially calcium signaling) plays a significant role in the pathogenesis of NSCL/P. Further genetic analyses identified potential pleiotropic loci primarily located at 16q24.2 and 3q21.1, involving key genes such as CASR and CSTA, and enriched in vitamin D response pathways. This study highlights the crucial position of genetic-environmental factors in the development of cleft lip and palate.

Objective To confirm whether PANoptosis is involved in the occurrence and development of non-syndromic cleft lip with or without cleft palate(NSCL/P)and identify key genetic variations and genes involved.Methods Firstly,a two-stage population was collected for genome-wide association study(GWAS)to determine the correlation between single nucleotide polymorphisms(SNPs)and NSCL/P.Next,the PANoptosis gene set was selected and SNPs were extracted to conduct quality control and gene-based associa-tion analysis in the stage Ⅰ population to screen candidate SNPs,and the stage Ⅱ samples were used as validation.The functional an-notation of Haploreg,RegulomeDB,and ATAC sequencing combined with linkage disequilibrium-based clumping further identified the key functional SNP and risk gene.Finally,expression quantitative trait loci(eQTL)analysis,RNA sequencing,single-cell sequen-cing,and protein-protein interaction analysis were used to predict the regulatory effects of the locus and gene.Results rs71518324 was associated with the risk of NSCL/P(Pmeta＜0.001),and the surrounding region was enriched with a large number of active func-tional element markers.The eQTL effect of rs71518324 on PRKAR1B gene was significant(P＜0.001).The expression of PRKAR1B in RNA sequencing generally increased over time.Five cell clusters were identified in single-cell RNA sequencing,with PRKAR1B mainly localized to the ectoderm.Conclusion rs71518324 in PRKAR1B,a PANoptosis related gene,is significantly associated with NSCL/P risk.

Objective:To systematically investigate the causal effects of exposure factors on nonsyndromic cleft lip with or without cleft palate (NSCL/P) using a phenome-wide mendelian randomization (MR-PheWAS) framework and identify pleiotropic loci.Methods:This study integrated genome-wide association study (GWAS) data for NSCL/P, including 1 069 cases and 1 724 controls, and systematically evaluated causal associations between exposures and NSCL/P using the MR-PheWAS framework. GWAS summary data for 2 106 Asian population-exposure phenotypes were obtained from the IEU OpenGWAS database. The inverse-variance weighted (IVW) method served as the core causal inference model, supplemented by weighted median and MR-Egger regression to verify the robustness of causal associations. Additionally, multivariable MR analysis was conducted to adjust for confounding effects, alongside sensitivity tests (Cochran′s Q and MR-PRESSO). Genetic correlations were analyzed using LD Score regression, and cross-phenotype pleiotropy analysis (PLACO/CPASSOC) was employed to identify shared genetic loci. Pathway enrichment and gene annotation data were integrated to explore potential biological mechanisms.Results:MR analysis identified serum calcium ( OR=0.12, P=0.019), high-density lipoprotein (HDL, OR=0.61, P=0.039), and mean corpuscular hemoglobin concentration (MCHC, OR=0.39, P=0.032) as protective factors, whereas serum sodium ( OR=21.41, P=0.013) was a risk factor. Furthermore, in subsequent analyses of genetic correlation and genetic overlap, a strong association was observed between serum calcium and NSCL/P. Cross-trait analysis localized pleiotropic loci to 16q24.2 and 3q21.1, involving CASR and CSTA, with significant enrichment in vitamin D response pathways. Conclusions:Numerous environmental exposure factors may have a causal impact on the outcomes of NSCL/P, and metabolic homeostasis (especially calcium signaling) plays a significant role in the pathogenesis of NSCL/P. Further genetic analyses identified potential pleiotropic loci primarily located at 16q24.2 and 3q21.1, involving key genes such as CASR and CSTA, and enriched in vitamin D response pathways. This study highlights the crucial position of genetic-environmental factors in the development of cleft lip and palate.

Objective To investigate the clinical phenotypic characteristics of a pedigree with congenital tooth agenesis(CTA)and identify the pathogenic gene using whole-exome sequencing(WES),aiming to confirm the disease-causing mutation site,explore its potential impact on protein structure and function,and provide new insights for the diagnosis of CTA.Methods The study focused on a pedigree with congenital absence of premolars.Blood samples were collected from pedigree members,and genomic DNA was extrac-ted.Potential pathogenic mutations were screened using WES and bioinformatics analysis.Candidate mutations were validated by Sanger sequencing.Gene Ontology(GO)functional annotation and KEGG pathway enrichment analysis were performed on co-expressed genes of the candidate gene.Results Clinical examination revealed that all four members of the family were patients with missing premolar teeth.WES identified two novel mutations in the TTN gene(c.94145G＞A and c.105406C＞T)in all affected family members.Sanger sequencing confirmed co-segregation of these mutations with the disease phenotype in the pedigree.Bioinformatics analysis indicated that Ttn was highly expressed during craniofacial development in mouse embryos.Enrichment analysis demonstrated that Ttn co-ex-pressed genes were significantly enriched in the extracellular matrix(ECM)receptor interaction and PI3K/Akt signaling pathway.Conclusion This study suggests that TTN is a potential pathogenic gene for congenital premolar agenesis in this pedigree.

Objective To confirm whether PANoptosis is involved in the occurrence and development of non-syndromic cleft lip with or without cleft palate(NSCL/P)and identify key genetic variations and genes involved.Methods Firstly,a two-stage population was collected for genome-wide association study(GWAS)to determine the correlation between single nucleotide polymorphisms(SNPs)and NSCL/P.Next,the PANoptosis gene set was selected and SNPs were extracted to conduct quality control and gene-based associa-tion analysis in the stage Ⅰ population to screen candidate SNPs,and the stage Ⅱ samples were used as validation.The functional an-notation of Haploreg,RegulomeDB,and ATAC sequencing combined with linkage disequilibrium-based clumping further identified the key functional SNP and risk gene.Finally,expression quantitative trait loci(eQTL)analysis,RNA sequencing,single-cell sequen-cing,and protein-protein interaction analysis were used to predict the regulatory effects of the locus and gene.Results rs71518324 was associated with the risk of NSCL/P(Pmeta＜0.001),and the surrounding region was enriched with a large number of active func-tional element markers.The eQTL effect of rs71518324 on PRKAR1B gene was significant(P＜0.001).The expression of PRKAR1B in RNA sequencing generally increased over time.Five cell clusters were identified in single-cell RNA sequencing,with PRKAR1B mainly localized to the ectoderm.Conclusion rs71518324 in PRKAR1B,a PANoptosis related gene,is significantly associated with NSCL/P risk.

Objective To study the three-dimensional changes of soft and hard tissue in male patients with unilateral non-syndromic cleft lip and palate in the mixed dentition period before and after maxillary protraction.Methods Twenty male patients with unilateral non-syndromic cleft lip and palate in the mixed dentition period treated by maxillary anterior traction in the Department of Orthodontics of Affiliated Stomatological Hospital of Nanjing Medical University were selected(average age(10.6±1.23)years old).Cone beam CT was taken before and after treatment.Dolphin 3D 11.95 software was used for three-dimensional measurement and analysis.SPSS 25.0 software package was used for statistical analysis.The self-controlled paired t test was used to compare the changes in soft and hard tis-sues of male patients with unilateral cleft lip and palate before and after treatment.The changes in the anterior displacement of the ANS point,the anterior displacement of point A,and the posterior displacement of point B were tested using the one-sample t test.Results The sagittal skeletal changes were significantly increased in ∠ SNA(P＜0.01),∠ANB(P＜0.01),Y axis(P＜0.05),the forward displacement of ANS point(P＜0.01)and A point(P＜0.01)and the backward displacement of B point(P＜0.01),but ∠SNB(P＜0.05)was decreased significantly.The vertical skeletal changes showed that ∠MP-FH(P＜0.01),∠MP-SN(P＜0.05)and the dis-tance of ANS-Me(P＜0.05)were increased significantly,but ∠SN-PP(P＜0.01)was decreased significantly.The dental changes inclu-ding ∠U1-NA(P＜0.01),the distance of U1-NA(P＜0.01),∠U1-SN(P＜0.01),overjet and the Wits were increased significantly,but ∠ L1-NB(P＜0.01),the distance of L1-NB(P＜0.01)and ∠L1-MP(P＜0.01)were decreased significantly.The changes of soft tis-sue including ∠ S-Ns-Sn(P＜0.01),∠ Sn-Ns-Bs(P＜0.01),the distance of UL-EP(P＜0.01)and LL-UL(P＜0.01)were increased sig-nificantly.Conclusion After the treatment of maxillary protraction,the forward growth of maxilla will be possibly promoted on patients with cleft lip and palate in the peak of growth timing,as well as the intermaxillary relationship and soft tissue profile,but the side effects should be paid attention to.

Objective To investigate the clinical phenotypic characteristics of a pedigree with congenital tooth agenesis(CTA)and identify the pathogenic gene using whole-exome sequencing(WES),aiming to confirm the disease-causing mutation site,explore its potential impact on protein structure and function,and provide new insights for the diagnosis of CTA.Methods The study focused on a pedigree with congenital absence of premolars.Blood samples were collected from pedigree members,and genomic DNA was extrac-ted.Potential pathogenic mutations were screened using WES and bioinformatics analysis.Candidate mutations were validated by Sanger sequencing.Gene Ontology(GO)functional annotation and KEGG pathway enrichment analysis were performed on co-expressed genes of the candidate gene.Results Clinical examination revealed that all four members of the family were patients with missing premolar teeth.WES identified two novel mutations in the TTN gene(c.94145G＞A and c.105406C＞T)in all affected family members.Sanger sequencing confirmed co-segregation of these mutations with the disease phenotype in the pedigree.Bioinformatics analysis indicated that Ttn was highly expressed during craniofacial development in mouse embryos.Enrichment analysis demonstrated that Ttn co-ex-pressed genes were significantly enriched in the extracellular matrix(ECM)receptor interaction and PI3K/Akt signaling pathway.Conclusion This study suggests that TTN is a potential pathogenic gene for congenital premolar agenesis in this pedigree.

Objective To study the distolingual space of mandibular molars in adult patients with different sagittal skeletal patterns,and to analyze the main bony anatomical sites that restrict molar distalization,in order to provide guidance for the treatment plan of mo-lar distalization.Methods A total of 97 adult patients according to the inclusion criteria were selected from the Department of Ortho-dontics,the Affiliated Stomatological Hospital of Nanjing Medical University.The patients were divided into skeletal Class Ⅰ group(n=28),skeletal Class Ⅱ group(n=49)and skeletal Class Ⅲ group(n=20)according to the ANB angle.CBCT of the patients were im-ported into Dolphin software for 3D reconstruction.The width of the distal root of the second molar,the width of alveolar bone,the dis-tance between the most convex point of the distal and lingual side of the distal root and the inneredge of the lingual cortex of the mandi-ble were measured at the 2,4,and 6 mm plane from the root furcation to the root apex.Statistical analysis was performed using SPSS 26.0 software,and univariate analysis of variance and LSD-t test were used to compare the difference.Results Root width was significantly narrower than alveolar bone width at all measurement planes(P＜0.01).Molar distolingual space in patients with different sagittal skeletal patterns was smaller than molar distal space,and the size of the space gradually decreased with the deepening of the measurement level,reaching the minimum value at the R4 and R6 measurement planes.Measurement results of this study showed that at the R6 level,the molar distolingual space in skeletal Class Ⅱ group was the minimum(2.30±2.45)mm;on the contrary,skeletal Class Ⅲ group was the maximum(4.17±2.38)mm.Conclusion When designing the plan of molar distalization in clinical practice,CBCT should be used,and more attention should be paid to the lingual alveolar bone mass of the mandibular molar.It is a safe and effective treatment method for skeletal Class Ⅰ and Ⅲ patients with mild to moderate dental crowding.

The biological samples of oral genetic diseases and rare diseases are extremely precious. Collecting and preserving these biological samples are helpful to elucidate the mechanisms and improve the level of diagnose and treatment of oral genetic diseases and rare diseases. The standardized construction of biobanks for oral genetic diseases and rare diseases is important for achieving these goals. At present, there is very little information on the construction of these biobanks, and the standards or suggestions for the classification and coding of biological samples from oral and maxillofacial sources, and this is not conducive to the standardization and information construction of biobanks for special oral diseases. This consensus summarizes the background, necessity, principles, and key points of constructing the biobank for oral genetic diseases and rare diseases. On the base of the group standard "Classification and Coding for Human Biomaterial" (GB/T 39768-2021) issued by the National Technical Committee for Standardization of Biological Samples, we suggest 76 new coding numbers for different of biological samples from oral and maxillofacial sources. We hope the consensus may promote the standardization, and smartization on the biobank construction as well as the overall research level of oral genetic diseases and rare diseases in China.