ObjectiveGenome-wide association studies have identified over 80 loci associated with nonsyndromic orofacial cleft (NSOC), yet substantial heritability remains unexplained. Insights from syndromic orofacial cleft (SOC) implicated genes could help bridge this gap.DesignA case-control association study in a Han Chinese cohort was conducted to evaluate the association between SOC-implicated genes and NSOC subtypes using association, linkage disequilibrium (LD), and haplotype analyses.SettingTertiary medical center.Patients, ParticipantsThe study included 1626 cases of non-syndromic cleft lip with or without cleft palate, 930 cases of non-syndromic cleft palate only, and 2255 controls.InterventionsPeripheral blood (cases) and umbilical cord blood (controls) were collected for DNA extraction.Main Outcome MeasuresAllelic (Pearson' s χ2, 1 df) and genotypic (Pearson' s χ2, 2 df) associations between SNPs and NSOC subtypes were evaluated, with odds ratios (ORs) and 95% confidence intervals (CIs). LD and sliding-window haplotype association analyses were performed in Haploview. SNPs with minor allele frequency (MAF) >0.05 and in Hardy-Weinberg equilibrium in controls were analyzed. The significance threshold was P < 1.27 × 10-5 after Bonferroni correction.ResultsAllelic analysis identified 23 SNPs that were significantly associated with NSOC subtypes (lowest P = 2.02 × 10-22). Genotypic analysis identified 39 significant SNPs (lowest P = 1.09 × 10-36). Signals mapped to 7 genes. Haplotype analyses revealed a shared causal variant block at MYMK and TWIST2, and allelic heterogeneity at NEDD4L.ConclusionsWe identified MYMK, TWIST2, and NEDD4L as NSOC-associated genes. Using SOC genes as prior knowledge reveals loci missed by standard GWAS, offering key insights into NSOC pathogenesis.

OBJECTIVE:To explore the association of PAX3 gene variants and non-syndromic orofacial cleft lip/palate (NSCL/P) among the Han Chinese population in Western China. METHODS:A total of 1,626 NSCL/P patients, 886 non-syndromic cleft palate only (NSCPO) pa-tients and 2,255 controls were recruited for association analysis. A total of 8 NSCPO patients, 7 non-syndromic cleft lip only (NSCLO) patients, and 3 normal controls were recruited for RNA sequencing. Blood samples were collected, genotype data for single nucleotide polymorphisms (SNPs) on the PAX3 gene were obtained and multi-level association analyses were conducted to explore their relationship with NSCL/P, including functional predictions of disease-related SNPs. Additionally, RNA sequencing was performed to investigate differences in PAX3 gene expression between patients with NSCL/P and controls. The study was conducted in PLINK analysis. RESULTS:rs2033806 was significantly associated with NSCL/P, whereas rs1430651 was mar-ginally associated with NSCL/P. None were significantly associated with NSCPO. In addition, rs2033806 was also associated with the subtypes NSCLP and NSCLO. CONCLUSION:rs2033806 at PAX3 showed different frequency distributions between non-syn-dromic oral cleft (NSOC) patients and controls, providing new insights into the aetiologies of NSOC.

OBJECTIVES:This study aims to explore the association between single nucleotide polymorphisms (SNPs) loci near the haplotype region hg19 chr9:100560865-100660865 of the forkhead box E1 (FOXE1) gene and the occurrence of non-syndromic cleft lip with or without cleft palate (NSCL/P) in western Han Chinese population. METHODS:In the first stage, our study recruited 159 NSCL/P patients and performed targeted region sequencing to screen SNPs loci near the haplotype region of the FOXE1 gene associated with NSCL/P. In the second stage, we selected 21 common SNPs and re-enrolled 1 000 non-syndromic cleft lip only (NSCLO) patients, 1 000 non-syndromic cleft palate only (NSCPO) patients, and 1 000 normal controls to verify the association. PLINK software was used to perform Hardy-Weinberg equilibrium (HWE) test. Association analysis for common variants, gene burden analysis for rare mutations, and function prediction of SNPs with non-synonymous mutations were performed using Mutation Taster and other software programs. RESULTS:In the first stage, 126 variants, including 76 single nucleotide variants and 50 insertion-deletions were identified. All the included SNPs confirmed to HWE, and the results of gene burden analysis and prediction of functional harmfulness for rare variants were not statistically significant. Association analysis showed that rs13292899 of the FOXE1 gene was significantly associated with NSCL/P (P=1.85E-27) and was also correlated with NSCLO (P=6.41E-23) and non-syndromic cleft lip with cleft palate (NSCLP) (P=2.36E-15) subtypes. In the validation phase, rs79268293 (P=0.013, P=0.022), rs10983951 (P=0.009 2, P=0.007 6), rs117227387 (P=0.009 2, P=0.007 6), rs3758250 (P=0.009 2, P=0.007 6), and rs116899397 (P=0.009 2, P=0.007 6) were significantly associated with NSCLO and NSCPO; rs13292899 (P=0.008 5), rs74606599 (P=0.008 3), rs143226042 (P=0.008 3), and rs117236550 (P=0.01) were associated with the occurrence of NSCLO; and rs12343182 (P=0.008 7), rs10119760 (P=0.012), rs10113907 (P=0.012), and rs13299924 (P=0.012) were associated with the occurrence of NSCPO. CONCLUSIONS:This study found a new susceptible SNP rs13292899 of the FOXE1 gene that is closely associated with NSCL/P and NSCLO subtype and 13 other SNPs associated with NSCLO or NSCPO.

ObjectivesNonsyndromic cleft lip with or without palate (NSCL/P) is one of the most prevalent congenital orofacial defects. It arises from a combination of genetic and environmental factors. This study aims to identify new risk loci around the musculoaponeurotic fibrosarcoma oncogene family, protein B (MAFB) gene in NSCL/P patients from the Western Han Chinese population.DesignA targeted region sequencing approach was employed to examine the MAFB gene in 159 NSCL/P cases. We conducted both single-variant association and gene-based burden analyses.SettingThe study was conducted in a stomatological hospital.Patients, participantsOne hundred and fifty-nine NSCL/P cases were analyzed.InterventionsBlood samples were collected.Main outcome measuresTo explore the association analysis between variants at MAFB and NSCL/P in Western Han Chinese population.ResultsWe identified a cluster of significant common variants near the 3' end of MAFB. Notably, rs6029223 showed significantly associated with NSCL/P (P = 3.82E-09, odds ration [OR] = 0.29, 95% confidence interval [CI]: 0.18-0.46), nonsyndromic cleft lip and palate (NSCLP) (P = 5.31E-08, OR = 0.25, 95% CI: 0.14-0.44) and nonsyndromic cleft lip only (NSCLO) (P = 1.3E-04, OR = 0.34, 95%CI: 0.19-0.62). In Addition, rs79836852 and rs200392238 were significantly associated with both NSCL/P and NSCLP.ConclusionsOur study revealed that single nucleotide polymorphisms near the 3' end of the MAFB gene are risk factors for NSCL/P and NSCLP in the Western Han Chinese population. Our findings reinforce the notion that MAFB is a susceptibility gene for NSCL/P.

The biological interactions between genetic and environmental modifiers play critical roles in the etiology of non-syndromic orofacial cleft (NSOC), but it is rarely studied. This study selected 47 environmental related genes from the metabolic pathways of smoking, drinking, hypoxia, and vitamins (including vitamin A, vitamin B9 (folic acid), vitamin C (ascorbic acid), vitamin D, and vitamin E), and test their associations with NSOC and its subtypes. We found that a novel gene SLC23A2, the vitamin C transporter gene is significantly associated with non-syndromic cleft palate only (NSCPO) (p = 3.25E-07, OR = 8.45, 95%CI:3.73-19.17). SLC23A2 is expressed in the craniofacial region of zebrafish (24hpf to 120hpf), obvious craniofacial abnormalities appeared in zebrafish (48hpf) when knock down the slc23a2 (slc23a2-MO). Knock down SLC23A2 in human embryonic palatal mesenchymal cell line (HEPM) induced decreased intracellular ascorbic acid (AA), increased reactive oxygen species (ROS), inhibited cell proliferation and triggered apoptosis, activated the PI3K-AKT-mTOR signaling pathway and inhibited autophagy; ROS levels and apoptosis ratio significantly decreased when we supplemented AA to HEPM cells with high ROS levels induced by Sin-1 (an exogenous ROS mimic). Knocked down SLC23A2 in HEPM cells or zebrafish, they became more sensitive to Sin-1, and AA supplementation was ineffective. In conclusion, we identified a novel susceptibility gene SLC23A2 for NSCPO, it may function by decreasing AA level, increasing the ROS levels, inducing apoptosis, and inhibiting autophagy through the activation of the PI3K-AKT-mTOR pathway in etiology of cleft palate.

Muenke syndrome is an autosomal dominant genetic disorder that is typically characterized by unilateral or bilateral coronal synostosis,macrocephaly,midface hypoplasia,and developmental delays.This article reports a case of Muenke syndrome with a soft cleft palate.A heterozygous missense mutation c.749C>G(p.P250A)was identified in the FGFR3 gene through genetic testing.The patient exhibited typical features including coronal synostosis,bilateral hearing loss,right accessory auricle,and developmental delays and underwent surgery to repair the soft cleft palate.Cases of Muenke syndrome with cleft palate in the literature are relatively rare,and common associated symptoms include coro-nal suture craniosynostosis and hearing impairment.This article reports a differential diagnosis with other craniosynosto-sis syndromes and provides a reference for clinical diagnosis and treatment.

Background Non-syndromic orofacial cleft (NSOC) is one of the most common craniofacial malformations with complex etiology. This study aimed to explore the role of specific SNPs in ZFP36L2 and its functional relevance in zebrafish models. Methods We analyzed genetic data of the Chinese Han population from two previous GWAS, comprising of 2512 cases and 2255 controls. Based on the Hardy-Weinberg Equilibrium (HWE) and minor allele frequency (MAF), SNPs in the ZFP36L2 were selected for association analysis. In addition, zebrafish models were used to clarify the in-situ expression pattern of zfp36l2 and the impact of its Morpholino-induced knockdown. Results Via association analysis, rs7933 in ZFP36L2 was significantly associated with various non-syndromic cleft lip-only subtypes, potentially conferring a protective effect. Zebrafish embryos showed elevated expression of zfp36l2 in the craniofacial region during critical stages of oral cavity formation. Furthermore, Morpholino-induced knockdown of zfp36l2 led to craniofacial abnormalities, including cleft lip, which was partially rescued by the addition of zfp36l2 mRNA. Conclusion Our findings highlight the significance of ZFP36L2 in the etiology of NSOC, supported by both human genetic association data and functional studies in zebrafish. These results pave the way for further exploration of targeted interventions for craniofacial malformations.

Non-syndromic orofacial clefts (NSOC) are common craniofacial birth defects, and result from both genetic and environmental factors. NSOC include three major sub-phenotypes: non-syndromic cleft lip with palate (NSCLP), non-syndromic cleft lip only (NSCLO) and non-syndromic cleft palate only (NSCPO), NSCLP and NSCLO are also sometimes grouped as non-syndromic cleft lip with or without cleft palate (NSCL/P) based on epidemiology. Currently known loci only explain a limited proportion of the heritability of NSOC. Further, differences in genetic susceptibility among the sub-phenotypes are poorly characterized. We performed a multi-ancestry GWAS meta-analysis on 44,094 individuals (9,381 cases, 28,510 controls, 2042 case-parent trios and 18 multiplex pedigrees) of East Asian, European, Latin and South American, and African ancestry for both NSOC and subtypes. We identified 50 loci, including 11 novel loci: four loci ( CALD1 , SHH , NRG1 and LINC00320 ) associated with both NSOC and NSCL/P, two loci ( NTRK1 and RUNX1 ) only associated with NSOC, four loci ( HMGCR , PRICKLE1 , SOX9 and MYH9 ) only associated with NSCL/P and one locus ( ALX1 ) specifically associated with NSCLO. Five of the novel loci are located in regions containing genes associated with syndromic orofacial clefts ( SHH , NTRK1, CALD1, ALX1 and SOX9 ); seven of the novel loci are located in regions containing genes-implicated in craniofacial development ( HMGCR, SHH, PRICKLE1, ALX1, SOX9, RUNX1, MYH9 ). Genetic correlation and colocalization analyses revealed an overlap between signals associated with NSCLO, NSCPO and NSCLP, but there were also notable differences, emphasizing the complexity of common and distinct genetic processes affecting lip and palate development.

Objective Paired box 7 (PAX7) has been considered as a candidate gene for non-syndromic cleft lip with or without palate (NSCL/P). However, there is no research for the XXX, and previous studies concentrated on limited variants. This study aimed to conduct sufficiently dense and powerful scans of variants at PAX7 and explored the roles of variants at PAX7 in NSCL/P among the XXX. Design Targeted region sequencing was performed to thoroughly screen variations, followed by a two-phase association analysis. 159 NSCL/P cases and 542 controls were analyzed in phase 1. Then in phase 2, the validation study was performed using 1626 cases and 2255 controls. We also explored the roles of variants at PAX7 gene in NSCL/P subtypes. Additionally, indirect associations were found by calculating LD and haplotypes. Setting The study was conducted in XXX. Patients, participants 159 NSCL/P cases and 542 controls were analyzed in phase 1. Then in phase 2, the validation study was performed using 1626 cases and 2255 controls. Interventions Blood samples were collected. Main outcome measures To explore the association analysis between variants at PAX7 and NSCL/P in XXX. Results The results showed that rs2236810, rs114882979 and rs2236804 were significantly associated with NSCL/P, which were predicted to have regulatory functions. Besides, variants at PAX7 function differently in the NSCL/P subtypes. We also discovered a PAX7 missense variant, NM_001135254 p.A369 V (NM_002584.2:c.1106C > T). Conclusions In summary, we confirmed 3 SNPs at PAX7 were significantly associated with NSCL/P in XXX and identified a missense variant, NM_001135254 p.A369 V (NM_002584.2:c.1106C > T).

Currently, no effective disc reconstruction treatment strategy is clinically available for temporomandibular joint (TMJ) disc-related diseases. To address this, we developed a prosthesis construct with laser-drilled decellularized natural disc reinforced by polycaprolactone, which mimics the natural morphology, and structural, biomechanical and biological property of the TMJ disc. The construct demonstrated good biocompatibility, safety and immunological tolerance both in vitro, and in a rat subcutaneous model. During 6 months implantation in an allogeneic rabbit TMJ disc reconstruction model, the disc prosthesis maintained its integrity, collagen fiber-orientation, mechanical property, joint structural stability and prevented articular cartilage and bone from damage. Furthermore, the "upgraded" disc prosthesis obtained from decellularized porcine disc was implanted into a goat TMJ disc reconstruction model. The xenograft prosthesis, with strength and viscoelasticity similar to a natural TMJ disc, was able to restore the structure and function of TMJ up to 20 weeks. These results demonstrate the translational feasibility of an allogeneic or xenogeneic decellularized disc prosthesis for treatment of advanced TMJ disc-related diseases. STATEMENT OF SIGNIFICANCE: This study makes a significant contribution to TMJ disc disease treatment both in theory and in clinics, because: (1) it provided an innovative approach to prepare an artificial TMJ disc with decent mechanical properties and long-term condyle-protecting effect; (2) it specified an advanced decellularized method for fibrocartilage decellularization and xenograft application; (3) it developed a facile and reproducible TMJ disc reconstruction model not only for middle size animal but also for large animal study; (4) the comprehensive and unreported biomechanical tests on the natural TMJ discs would act as a valuable reference for further research in the field of artificial TMJ disc materials or TMJ disc tissue engineering; (5) it suggested a potential treatment for patients with severe TMJ diseases that were commonly met but difficult to treat in clinics.

Objectives Non-syndromic cleft palate only (NSCPO) is a common congenital deformity with complex etiologies. GRHL3, FAF1, and KCNJ2 have been reported to be involved in the pathogenesis of NSCPO. Up till now, there have been no replication studies based on large Han Chinese. Therefore, this study aimed to investigate associations between GRHL3, FAF1, KCNJ2, and NSCPO sub-phenotypes patients in Han Chinese. Materials and Methods Firstly, we selected 2 SNPs based on previous literatures: FAF1 (rs3827730) and GRHL3 (rs41268753). Also, we selected 8 tagSNPs in GRHL3 (rs557811, rs609352, rs10903078, rs6659209, rs12401714, rs12568599, rs3887581, rs12024148) and 2 tagSNPs in KCNJ2 (rs75855040 and rs236514). Afterward, we evaluated these SNPs among 1668 NSCPO patients and 1811 normal controls from Han Chinese. Following data were analyzed by PLINK and Haploview program. Results Association analysis under additive model showed that allele A at rs12568599 in GRHL3 gene is significantly associated with NSCPO (p = 0.0034, OR = 1.38 and 95%CI: 1.11-1.72) and its sub-phenotype incomplete cleft palate (ICP) (p = 0.0039, OR = 1.4 and 95%CI: 1.11-1.75), and it could increase the risk of both NSCPO and ICP. Conclusions This study firstly found that rs12568599 in GRHL3 is associated with NSCPO and ICP in Han Chinese, indicating that sub-phenotypes of NSCPO have different etiologies.

The purpose of this study is to analyze the clinical characteristics of a Treacher Collins syndrome (TCS) patient carrying a de novo variant of TCOF1, and briefly analyze the correlation between genetic results and clinical features. Also, the pathogenesis and clinical treatment of TCS are reviewed. A Chinese pedigree with TCS containing 8 members was enrolled. Phenotype of the proband was evaluated by a surgeon, then whole exome sequencing of the proband was performed. Then we verified the proband-derived variants by Sanger sequencing in the pedigree. Correlation between genotype and phenotype was analyzed. The study was conducted in a stomatological hospital. A Chinese pedigree with TCS containing 8 members. To ascertain the genetic variants in the Chinese pedigree with TCS. Blood samples were collected. We reported a case of typical TCS with a de novo missense variant (NM_001371623.1:c.38T>G, p.(Leu13Arg)) in exon 1 of TCOF1, who presented asymmetrical facial abnormalities, including downward slanting of the palpebral fissures, sparse eyebrows, lateral tilt of the eyeballs, bilateral external ears deformities, hypoplasia of midface, reduction of the zygomatic body, bilateral orbital invagination, right external auditory canal atresia, mandibular ramus short deformity, cleft palate and the whole face was convex. This research found a novel variant of TCS in Chinese, expanding the spectrum of TCS pathogenic variants. Genetic results combined with clinical phenotype can make a definite diagnosis and provide genetic counseling for the family.

唇腭裂是最常见的先天畸形之一.根据是否伴有全身其他部位的先天疾病,可分为非综合征型唇腭裂和综合征型唇腭裂.研究发现非综合征型唇腭裂为多因素遗传病,同时受多种遗传和环境危险因素的影响.而综合征型唇腭裂人群发病率相对较低,为单基因病,符合孟德尔遗传定律,呈明显的家族遗传性,且大多有明确的致病基因.本文将就5种发病率较高,较为常见的综合征型唇腭裂近年来遗传学方面的进展进行介绍,为将来综合征型唇腭裂遗传研究提供参考.

Objective To explore the changes of T-lymphocy te subsets in peripheral blood and immunophenotype features in bone marrow of the patients with mutiple myeloma(MM). Methods The immuophenotype in bone marrow and the T-lymphocyte subsets in peripheral blood of 32 MM patie nts were detected by flow cytometry (FCM). Results In preth erapy stage and developing stage,the percentages of CD3,CD4 and the ratios CD4/C D8 were decreased, while the percentags of CD8 were increased.The percentages of CD3,CD4, CD8, and the ratio CD4/CD8 in stable stage were normal.To compare wit h normal controls, the percentage of NK cells in pretherapy stage was increased, but there was no obvious changes in stable stage and developing stage.The posit ive rate of CD56,CD54,CD38,CD19 or CD20 in pretherapy stages was 89.4%,100%,100% ,15.8%, and 5.35%,respectively.The positive rate of CD56,CD54,CD38,CD19 or CD2 0 in developing stages was 100%,100%,100%,20%, and 40%,respectively.The positiv e rate of CD56,CD54 or CD38 in stable stage was 37.5 %,100%, and 100%,respec tively, while there were no expression of CD19 and CD20.There were a few express ions of the markes of T-cell in pretherapy stage. Conclusion Immuophenotype analysis in bone marrow and detection of T-lymphocyte subsets in peripheral blood are helpful to the diagosis and monitoring treatment for MM patients.