Research progress on the regulation of craniofacial development and malformation by fibroblast growth factor 8 / 口腔疾病防治

@#Fibroblast growth factor 8 (FGF8) is a kind of secretory polypeptide that has crucial roles in the development of various tissues and organs. Current studies have found that FGF8 can regulate the differentiation of cranial neural crest cells by activating the mitogen-activated protein kinase (MAPK) signaling pathway and affect the establishment of mandibular arch polarity and the development of craniofacial symmetry by regulating the expression of target genes. Cleft lip with or without cleft palate, ciliopathies, macrostomia and agnathia are four developmental malformations involving the craniofacial region that seriously affect the quality of life of patients. The abnormal FGF8 signal caused by gene mutation, abnormal protein conformation or expression is closely related to the occurrence of craniofacial malformations, but the molecular mechanism and signaling pathway underlying these malformations have not been fully elucidated. Craniofacial development is a complex process mediated by a variety of signaling molecules. In the future, the role of various signaling molecules in craniofacial development and malformations need to be explored to provide a new perspective and vision for the prevention and treatment of these craniofacial malformations.

Composición y función salival en niños con labio con/sin paladar hendido no sindrómico / Salivary composition and function in children with non-syndromic cl w/wo p / Composição e função salivar em crianças com fissuras labiais e/ou palatinas não sindrômicas

Resumen Introducción y Objetivo: La saliva cumple una importante función en la homeostasis oral, aunque no es evaluada rutinariamente en pacientes con labio con/sin paladar hendido (LH±PH). Este estudio pretendió comparar las características salivares de niños con LH±PH no sindrómico vs. niños sin esta diferencia craneofacial. Materiales y métodos: Se incluyeron 17 niños con LH±PH no sindrómico y 25 niños sin LH±PH, de los cuales se colectaron muestras de saliva estimulada (SE) y no estimulada (SNE). Se realizaron pruebas fisicoquímicas, cuantificación de iones (Ca, Mg y P), proteína total y recuento de streptococcus mutans en ambos grupos para ambas muestras de saliva. Para establecer comparaciones entre grupos se realizó la prueba no paramétrica de Mann-Whitney. Resultados: Hubo diferencias estadísticamente significativas en el pH de SE y SNE, así como en proteína total y capacidad amortiguadora de SNE. El pH (p=0,002) y capacidad amortiguadora (p=0,023) de SNE de los niños con LH±PH fueron menores, mientras que la concentración de proteína total (p=0,003) fue mayor con respecto al grupo control. Se encontró mayor proporción de estreptococo mutans en SNE de niños con LH±PH (6,39x104 UFC mL-1) respecto al grupo control (2,21x104 UFC mL-1), aunque sin detectar diferencias estadísticamente significativas (p=0,243). Conclusiones: Estos hallazgos sugieren una alteración en la función y composición de la SNE en pacientes con LH±PH no sindrómico, que correspondería a una alteración en la formación y función de las glándulas salivales en estos individuos que requiere ser investigada.

Abstract Introduction and objective: Saliva plays an important role in oral homeostasis, although it is not routinely evaluated in patients with CL±P. This study aimed to compare these characteristics in children with non-syndromic CL±P vs. children without this craniofacial difference. Materials and methods: This descriptive study included 17 children with non-syndromic CL±P from Fundación Clínica Noel and 25 children without CL±P. Stimulated (SS) and non-stimulated saliva (NSS) from both groups were collected for physicochemical tests, quantification of ions (Ca, Mg and P), total protein and streptococcus mutans count. Results: There were statistically significant differences in pH of SS and NSS, as well as in total protein concentration and buffer capacity in NSS. The pH (p = 0.002) and buffering capacity (p = 0.023) of NSS of CL±P children decreased, while total protein concentration (p = 0.003) increased compared to the control group. A higher streptococcus mutans count was found in NSS of CL±P children (6.39x104 CFU UFC mL-1) compared to the control group (2.21x104 UFC mL-1), although without detecting statistically significant differences (p = 0.243). Conclusions: These findings suggest an alteration in the function and composition of NSS in patients with non-syndromic CL±P; which could possibly be explained from the genetic/biological point of view by an alteration in the salivary glands formation and function in these individuals that requires further investigation.

Resumo Introdução e objetivo: A saliva desempenha um papel fundamental na homeostase oral, não é avaliada rotineiramente em pacientes com FL/P. Portanto, este estudo objetivou indagar essas características em crianças com FL/P não sindrômico. Materiais e métodos: 17 crianças com FL/P não sindrômico da Fundación Clínica Noel, e 25 crianças sem FL/P foram incluídas neste estudo. Amostras de saliva estimulada (SE) e saliva não estimulada (SNE) foram coletadas para realização de testes físico-químicos, quantificação de íons (Ca, Mg e P), proteína total e contagem de streptococcus mutans. Resultados: Houve diferenças estatisticamente significativas no pH de SE e SNE, bem como na concentração de proteína total e capacidade tampão de SNE. O pH (p=0,002) e a capacidade tampão (p=0,023) de SNE de crianças com FL/P foram menores, enquanto a concentração de proteína total (p=0,003) foi maior, quando comparado ao grupo controle. Um contagem maior de streptococcus mutans foi encontrada na saliva não estimulada de crianças com FL/P (6,39x104 UFC mL-1) em comparação ao grupo controle (2,21x104 UFC mL-1), embora sem detectar diferenças estatisticamente significativas (p=0,243). Conclusão: Esses achados sugerem uma alteração na função e composição da SNE em pacientes com FL/P não sindrômico; isso possivelmente poderia ser explicado do ponto de vista genético/biológico por uma alteração na formação e função das glândulas salivares nesses indivíduos que requer investigação.

Association between the JAG2 gene polymorphism and the occurrence of nonsyndromic cleft lip with or without cleft palate in northwest Chinese population / 中华医学遗传学杂志

Objective@#To assess the association of JAG2 gene single nucleotide polymorphisms with the occurrence of nonsyndromic cleft lip with or without cleft palate (NSCLP) among northwest Chinese population.@*Methods@#A case-control study was carried out on 301 NSCLP patients and 304 healthy controls. An iMLDR™ genotyping technique was used to detect three single nucleotide polymorphisms (SNPs) [rs741859 (T/C), rs11621316 (A/G) and rs1057744(C/T)] of the JAG2 gene. Allelic and genotypic frequencies and haplotypic distribution among the two groups were compared.@*Results@#A significant difference was found in the frequency of C and T alleles for rs741859 between the two groups. The CT genotype of rs741859 could significantly reduce the risk for NSCLP to 65% (P < 0.05) and the risk for cleft lip with or without cleft palate (CL/P) to 62% (P < 0.05). rs11621316 and rs1057744 are in the same linkage disequilibrium (LD) region with a high degree of linkage (r2 > 0.8), whose distribution difference between the two groups was not statistically significant (P > 0.05).@*Conclusion@#The CT genotype of the JAG2 gene rs741859 may confer a protective effect for NSCLP among northwest Chinese population.

Exploring parent-of-origin effects for non-syndromic cleft lip with or without cleft palate on PTCH1, PTCH2, SHH, SMO genes in Chinese case-parent trios / 北京大学学报(医学版)

OBJECTIVE@#Non-syndromic cleft lip with or without cleft palate (NSCL/P) is a common birth defect, affecting 1.4 per 1 000 live births, and multiple genetic and environmental risk factors influencing its risk. All the known genetic risk factors accounted for a small proportion of the heritability. Several authors have suggested parent-of-origin effects (PoO) may play an important role in the etiology of this complex and heterogeneous malformation. To clarify the genetic association between PTCH1, PTCH2, SHH and SMO in hedgehog (HH) pathway and NSCL/P, as well as testing for potential PoO effects in Chinese case-parent trios.@*METHODS@#We tested for transmission disequilibrium tests (TDT) and PoO effects using 83 common single nucleotide polymorphic (SNP) markers of HH pathway genes from 806 NSCL/P case-parent trios. These trios were drawn from an international consortium established for a genome-wide association studies (GWAS) of non-syndromic oral clefts of multiple ethnicities. DNA samples were collected from each trio. Single marker and haplotype based analysis were performed both in TDT tests and PoO effects. SNPs were excluded if they (ⅰ) had a call rate of < 95%, (ⅱ) had a minor allele frequency (MAF) of < 0.05, (ⅲ) had Mendelian errors over all trios of >5%, (ⅳ) had a genotype distribution in the parents that deviated from the Hardy-Weinberg equilibrium (HWE) (<i>P</i> < 0.000 1). The process was done using Plink (version 1.07, <a href="http://pngu.mgh.harvard.edu/~purcell/plink/data.shtml" target="_blank">http://pngu.mgh.harvard.edu/~purcell/plink/data.shtml</a>). TDT test was performed in Plink v1.07. A log-linear model was used to explore PoO effects using Haplin v6.2.1 as implemented in R package v3.4.2. Significance level was assessed using the Bonferroni correction.@*RESULTS@#A total of 18 SNPs were dropped due to low MAF, thus leaving 65 SNPs available for the analysis. Thus the Bonferroni threshold was 7.7×10-4 (0.05/65). Nominal significant association with NSCL/P was found at a SNP (rs4448343 in PTCH1, P=0.023) and six haplotypes (rs10512249-rs4448343, rs1461208-rs7786445, rs10512249-rs4448343, rs16909865-rs10512249-rs4448343, rs1461208-rs7786445-rs12698335, and rs288756-rs288758-rs1151790, P < 0.05). A total of six haplotypes (rs288765-rs1233563, rs12537550-rs11765352, rs872723-rs288765-rs1233563, rs288765-rs1233563-rs288756, rs6459952-rs12537550-rs11765352, and rs12537550-rs11765352-rs6971211) showed PoO effect (P < 0.05). None of the results remained significant after the Bonferroni correction (P>7.7×10-4).@*CONCLUSION@#Neither significant association between SNPs within HH pathway and the risk of NSCL/P nor PoO effects was seen in this study.

Association between platelet-derived growth factor-C single nucleotide polymorphisms and nonsyndromic cleft lip with or without cleft palate in Western Chinese population / 华西口腔医学杂志

OBJECTIVE@#To explore the association between two single nucleotide polymorphisms (SNPs), namely, rs4691383 and rs7667857, in the platelet-derived growth factor-C (PDGF-C) gene, the genotypes, environmental exposure factors, and nonsyndromic cleft lip with or without cleft palate (NSCL/P) in Western Chinese population.@*METHODS@#A total of 268 case-parent trios were selected, and two SNPs (rs4691383 andrs7667857) were genotyped by using polymerase chain reaction and restriction enzyme fragment length polymorphic method and direct sequencing method. Hardy-Weinberg equilibrium, linkage disequilibrium test, transmission disequilibrium test, and haplotype analysis were conducted to analyze the data. Meanwhile, the questionnaires on the epidemiology of cleft lip and palate filled by the included samples were collected, and the interaction between the genotypes of the two SNPs and environmental exposure factors was assessed by conditional logistic regression.@*RESULTS@#The A allele at rs4691383 and the G allele at rs7667857 of PDGF-C gene were over-transmitted for NSCL/P (P0.05).@*CONCLUSIONS@#The rs4691383 and rs7667857 at PDGF-C gene are closely related to the occurrence of NSCL/P in Western Chinese population. However, the interaction between environmental exposure factors and PDGF-C genotypes is not obvious in the occurrence of NSCL/P.

Associations among PRDM16 polymorphisms, environmental exposure factors during mother's pregnancy, and nonsyndromic cleft lip with or without cleft palate / 华西口腔医学杂志

OBJECTIVE@#We aimed to study the association between rs7525173, rs2236518, rs2493264 single nucleotide polymorphism (SNP) in the PRDM16 gene, smoking, alcohol exposures, and nonsyndromic cleft lip with or without cleft palate (NSCL/P).@*METHODS@#A total of 157 case-parent trios were selected, and SNPs were genotyped by using ligase detection reaction (LDR) and direct sequencing methods. Transmission disequilibrium test (TDT) and linkage disequilibrium (LD) tests were con-ducted to analyze the data. A total of 1 710 patients with orofacial clefts and 956 healthy newborns were enrolled in the epidemiological survey. The smoking and drinking exposures of parents during early pregnancy were analyzed.@*RESULTS@#The C allele at rs2236518 was over-transmitted for NSCPO (P<0.05). Statistical differences were observed among three factors, namely, maternal smoking, maternal passive smoking, and maternal drinking (P<0.05).@*CONCLUSIONS@#The rs2236518 at PRDM16 gene, maternal smoking, maternal passive smoking, and maternal drinking were closely related to the occurrence of NSCL/P.

Association of single nucleotide polymorphisms in AXIN2, BMP4, and IRF6 with Non-Syndromic Cleft Lip with or without Cleft Palate in a sample of the southeast Iranian population

Abstract Non-syndromic cleft lip with or without palate (NSCL/P) is a common congenital malformation worldwide, with complex etiology. It has been proposed that interaction of genes and environmental factors play a role in the predisposition to this disease. Objectives: The aim of this study was to examine the association between AXIN2 (axis inhibition protein 2) rs7224837, BMP4 (bone morphogenetic protein 4) rs17563, and IRF6 (interferon regulatory factor 6) rs861019 and 2235371 polymorphisms and NSCL/P in an Iranian population. Material and Methods: This case-control study was carried out on 132 unrelated NSCL/P patients and 156 healthy subjects. The variants were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The findings suggest that BMP4 rs17563 polymorphism significantly decreased the risk of NSCL/P in codominant (OR=0.36, 95%CI=0.17-0.79, p=0.012, CT vs CC and OR=0.11, 95%CI=0.01-0.88, p = 0.019, TT vs CC), dominant (OR=0.30, 95%CI=0.15-0.62, p = 0.0007, CT+TT vs CC), recessive (OR=0.12, 95%CI=0.02-0.99, p = 0.023, TT vs CC+CT), overdominant (OR=0.39, 95%CI = 0.18-0.84, p=0.021, CT vs CC+TT), and allele (OR=0.28, 95%CI=0.15-0.55, p<0.0001, T vs C) inheritance models. Our findings did not support an association between AXIN2 rs7224837 and IRF6 rs861019 polymorphism and risk/protection of NSCL/P. The IRF6 2235371 variant was not polymorphic in our population. Conclusion: The results indicate that the BMP4 rs17563 variant is likely to confer a protective effect against the occurrence of NSCL/P in a sample of the southeast Iranian population.

Association between non-syndromic cleft lip with or without cleft palate and environmental factors in Ningxia / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the association between non-syndromic cleft lip with or without cleft palate (NSCL/P) and environmental factors in Ningxia population.</p><p><b>METHODS</b>This case-control study involved 453 NSCL/P patients and 452 normal newborns from Ningxia. A questionnaire focusing on various factors, including family history, pregnancy reaction, drug use during pregnancy, and infections, was used and responses were analyzed through Chi-square test and Logistic regression analysis with SPSS 16.0.</p><p><b>RESULTS</b>The constituent ratio of different types of NSCL/P was cleft lip∶cleft lip and palate∶cleft palate equal to 1︰2.02︰1.51. Logistic regression analysis revealed that abnormal pregnancy, infection, abortion, drugs, drinking, smoking, and living near factories likely increased the risk of NSCL/P (P<0.05). Single fetus, pregnancy-related nausea, vomiting, parents' moderate tastes, and eating soy foods and fruits decreased the risk of NSCL/P (P<0.05).</p><p><b>CONCLUSIONS</b>The incidence of NSCL/P should be reduced to enhance the conditions of women during pregnancy by maintaining a balanced diet and avoiding infections, abortion, drugs, and negative habits. 
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Long-Term Follow-Up Study of Young Adults Treated for Unilateral Complete Cleft Lip, Alveolus, and Palate by a Treatment Protocol Including Two-Stage Palatoplasty: Speech Outcomes

BACKGROUND: No consensus exists on the optimal treatment protocol for orofacial clefts or the optimal timing of cleft palate closure. This study investigated factors influencing speech outcomes after two-stage palate repair in adults with a non-syndromal complete unilateral cleft lip and palate (UCLP). METHODS: This was a retrospective analysis of adult patients with a UCLP who underwent two-stage palate closure and were treated at our tertiary cleft centre. Patients ≥17 years of age were invited for a final speech assessment. Their medical history was obtained from their medical files, and speech outcomes were assessed by a speech pathologist during the follow-up consultation. RESULTS: Forty-eight patients were included in the analysis, with a mean age of 21 years (standard deviation, 3.4 years). Their mean age at the time of hard and soft palate closure was 3 years and 8.0 months, respectively. In 40% of the patients, a pharyngoplasty was performed. On a 5-point intelligibility scale, 84.4% received a score of 1 or 2; meaning that their speech was intelligible. We observed a significant correlation between intelligibility scores and the incidence of articulation errors (P<0.001). In total, 36% showed mild to moderate hypernasality during the speech assessment, and 11%–17% of the patients exhibited increased nasalance scores, assessed through nasometry. CONCLUSIONS: The present study describes long-term speech outcomes after two-stage palatoplasty with hard palate closure at a mean age of 3 years old. We observed moderate long-term intelligibility scores, a relatively high incidence of persistent hypernasality, and a high pharyngoplasty incidence.

Association of rs1 1 742688 polymorphism of DHFR gene with non-syndromie cleft lip with or without cleft palate in the northeast of China / 实用口腔医学杂志

Objective:To investigate the association between dihyrofolate reductase(DHFR)gene rs1 1 742688 polymorphism and non-syndrom cleft lip with or without cleft palate (NSCL/P)in northest Chinese population.Methods:PCR-restriction fragment length polymorphism(PCR-RFLP)was used to identify the rs1 1 742688 polymorphism of DHFR gene of 220 NSCL/P patients(inclu-ding 1 38 core families)and 1 80 healthy controls.Hardy-Weinberg test and SPSS statistical software were used to calculate the data, OR and 95% confidence intervalarents.Results:In case-contral analysis,there was no significant difference in TT genotype of rs1 1 742688 between NSCL/P subjects and the controls(χ2 =0.439,P >0.05)in.Conclusion:The polymorphism of rs1 1 742688 in DHFR gene is not associated with NSCL/P in northest Chinese population.

Research progress on susceptible gene loci of non-syndromic cleft lip with or without deft palate / 国际儿科学杂志

Non-syndromic cleft lip with or without cleft palate is a common birth defect with complex causes,now widely considered as the result of interactions between genetic and environmental factors.Susceptible genes of congenital cleft lip with or without cleft palate have become a focus since the whole genome sequencing was available.Numerous candidate genes which were screened out are being constantly validated by gene polymorphism detection,case-control study and meta analysis.But the results are inconsistent.In this article,we review the research progress on susceptible genes of non-syndromic cleft lip with or without cleft palate and the relationship between environmental factors and cleft lip with or without palate in recent years.

The 19-bp deletion polymorphism of dihydrofolate reductase (DHFR) and nonsyndromic cleft lip with or without cleft palate: evidence for a protective role

Objective Nonsyndromic cleft lip with or without cleft palate (NS-CL/P) are among the most common congenital birth defects worldwide. Several lines of evidence point to the involvement of folate, as well as folate metabolizing enzymes in risk reduction of orofacial clefts. Dihydrofolate reductase (DHFR) enzyme participates in the metabolic cycle of folate and has a crucial role in DNA synthesis, a fundamental feature of gestation and development. A functional polymorphic 19-bp deletion within intron-1 of DHFR has been associated with the risk of common congenital malformations. The present study aimed to evaluate the possible association between DHFR 19-bp deletion polymorphism and susceptibility to NS-CL/P in an Iranian population. Material and Methods The current study recruited 100 NS-CL/P patients and 100 healthy controls. DHFR 19-bp deletion was determined using an allele specific-PCR method. Results We observed the DHFR 19-bp homozygous deletion genotype (D/D) vs. homozygous wild genotype (WW) was more frequent in controls than in NS-CL/P patients (25% vs. 13%), being associated with a reduced risk of NS-CL/P in both codominant (OR=0.33, P=0.027) and recessive (OR=0.45, P=0.046) tested inheritance models. We also stratified the cleft patients and reanalyzed the data. The association trend for CL+CL/P group compared to the controls revealed that the DD genotype in both codominant (OR=0.30, P=0.032) and recessive models (OR=0.35, P=0.031) was associated with a reduced risk of CL+CL/P. Conclusions Our results for the first time suggested the DHFR 19-bp D/D genotype may confer a reduced risk of NS-CL/P and might act as a protective factor against NS-CL/P in the Iranian subjects. .

The association of MAFB gene polymorphism with nonsyndromic cleft lip with or without cleft palate in Ningxia Hui and Han population / 实用口腔医学杂志

Objective:To study the association between non-syndrome cleft lip and palate (NSCL/P)and MAFB (rs6072081, rs6065259,rs17820943,rs11696257)gene polymorphism in NingXia Hui and Han population.Methods:512 cases of NSCL/P,258 case of Hui nationality and 254 of Han,174 cases of complete 3 core family members(patients and their parents),142 cases of single parents families(patients and their single parents),were collected in patient group.505 cases of healthy newborns were collected in control group.The MAFB gene SNPs were determined with TaqMan SNPs genotyping methods and the data were analyzed by case-con-trol analysis,transmission disequilibrium test(TDT)and family based association test(FBAT).Results:Case-control analysis found that,there was a statistical significance of the genotypes and allele frequency between patients and the controls in MAFB of the 4 locus (P 0.05).Conclusion:The 4 locus single nucle-otide polymorphism was associated with NSCL/P in Ningxia population.

Association between TGF-α gene BaraH Ⅰ polymorphism and nonsyndromic cleft lip with or without cleft palate / 中华医学美学美容杂志

Objective Nonsyndromic cleft lip with or without cleft palate(NSCL/P)is a common craniofacial birth defect which results in lifelong medical and social consequences.Although Asians have the highest birth prevalence of oral-facial clefts,the majority of gene mapping studies of cleft lip with or without cleft palate(CL/P)have been in European or Ameriean Caucasians.Therefore,the obiective of this study was to evaluate association between transforming growth factor alpha(TGF-α)gene BamH Ⅰ polymorphism and NSCL/P in Chinese.Methods 107 patients with NSCL/P and 136 healthy controls were examined for TGF-α/BamH Ⅰ genotypes.TGF-α/BamH Ⅰ typing was carried out by digesting the locus specific polymerase chain reaction amplified products with alleles specific BamH Ⅰ restriction enzyme(PCR-RELP).Resuits A1 allele frequency was 0.06 and A2 allele frequency was 0.94 in the controls.A1 allele frequency was 0.14 and A2 allele frequency was 0.86 in patients with NSCL/P(x2=8.27,df=1,P＜0.05).A1 allele frequency was 0.17 and A2 allele frequency was 0.83 in the bilateral cleft lip with or without cleft palate.A1 allele frequency was 0.13 and A2 allele frequency was 0.87 in the unilateral cleft lip with or without cleft palate(x2=0.36,df=1,P＞0.05).There was no statistically significant between the case with family history and the case without family history(x2=0.34,df=1,P＞0.05).Conclusions The above data demonstrate that there is evidence for the association of TGF-α polymorphism with development of NSCL/P in Chinese.

Association of some SNP in TGFA,IRF6 and nonsyndromic cleft lip with or without cleft palate by DNA microarray technology / 中华检验医学杂志

Objective To explore the relationship between some single nucleotide polymorphisms (SNP)loci of interferon regulatory factor 6(IRF6)gene,transforming growth faetor-?(TGFA)gene and nonsyndromic cleft lip with or without cleft palate(NSCL/P)in nuclear families consisting of fathers, mothers and affected offspring with NSCL/P from southeast China.Methods Some SNloci of IRF6 and TGFA were detected by applying microarray technology in nuclear families,and then haplotype relative risk (HRR)and transmission disequilibrium test(TDT)were performed.Results There were no significant difference in genotypes and alleles distribution between patients and their parents.The SNP locus——V274I of IRF6 was associated with NSCL/P(HRR:?~2=4.5816,P