目的
1. 通过Linked-Read测序全基因组分析非综合征型唇腭裂家系中的遗传信息，寻找与该非综合征型唇裂家系相关的遗传变异。
2.1通过对采用阶梯状Le Fort I 型截骨利用下颌骨外板植骨手术患者的随访研究，探索该手术方式是否能降低唇腭裂正颌患者术后的复发率。
2.2通过对采用二阶段牵引成骨（上颌牵引成骨+上颌即刻坚固内固定）患者的随访分析，初步了解二阶段牵引成骨的复发率和采用改良外置式牵引支架中发生的临床问题。
方法
1. 以非综合征型唇腭裂家系为研究对象，使用Linked-Read测序技术获取患者及其家庭相关成员的全基因组遗传变异信息。与人类基因组参考序列hg38进行比对，通过Long Ranger软件调用BWA、GATK等检测模块获取变异信息；利用ANNOVAR对检测出的变异位点进行功能注释；再根据突变所在的位置、位点的保守性及危害度、等位基因频率、与颅面部发育的相关性、与唇腭裂的相关性等进一步筛选并获得与非综合征型唇腭裂相关的遗传变异。
2.1回顾性分析了58位上颌后缩前移距离在6-9mm之间的患者。其中实验组28位采用下颌骨外板植骨，对照组30位采用常规Le Fort I 型截骨。术后随访12个月，分别在术前术后拍摄头颅侧位片。通过投影测量分析实验组与对照组术前术后上颌骨距离与角度的变化，比较两组复发率是否存在差异。
2.2回顾性分析了11例采用二阶段牵引成骨的单侧唇腭裂患者。术后随访12个月，分别于术前、牵引完成、上颌坚固内固定术后2周和术后1年拍摄头颅侧位片，用以分析上下颌骨在距离和角度上的变化情况。其中5名患者在前方牵引的过程中配合横向扩弓的治疗，于术前和术后固定结束时拍摄三维CT，分析扩弓治疗的效果。同时对患者牵引过程中出现的临床问题和术后语音功能的变化做了相应的分析和记录。
结果
1.本研究通过对非综合征型唇腭裂家系的Linked-Read全基因测序分析，发现了与该家系非综合征型唇腭裂发生相关的变异位点和拷贝数变异，即AXIN2（c.2018G＞A, p.S673F）和155kb的ADAM3A/ADAM5片段（chr8, 39374555-39529710）。
2.1实验组上颌前移7.13±0.7mm（范围：6.01-8.23mm），术后1年平均复发率为25.07±6.64%。对照组上颌前移6.90±0.55mm（范围：6.05-7.39mm），术后1年平均复发率为24.89±4.25%。两组之间复发率没有显著性差异。
2.2 11名患者上颌骨在牵引完成时上颌骨平均向前移动13.9mm，向下移动7.7mm；下颌骨平均向后移动5.3mm，向下8.5mm。上颌骨和下颌骨出现了明显的顺时针旋转。患者出现了明显的张口受限。上颌坚固内固定后与术前相比，上颌骨平均向前移动10.1mm， 向下移动3.4mm；下颌骨平均向后移动2.4mm，向下4.7mm。患者最大张口度正常。术后1年患者上颌骨水平向复发率为22.2%，垂直向复发率为51.6%。患者张口受限症状恢复。5名患者横向扩弓治疗后，牙弓宽度增加明显，尖牙牙冠之间宽度平均增加6.4mm，第一磨牙近中颊尖间的牙冠宽度平均增加6.2mm。术后没有出现明显的语音功能恶化的情况。
结论
1. 本研究发现了AXIN2基因和ADAM3A/ADAM5片段为该非综合型唇腭裂家系的易感基因和易感片段，为阐明唇腭裂的发病机制提供了新的遗传学依据。
2.1在采用阶梯状Le Fort I型截骨上颌骨需要前移6-9mm的单侧唇腭裂患者中，采用下颌骨外板移植并不能有效减小上颌骨的复发率。
2.2 采用二阶段牵引成骨的手术方法能在有效前移上颌骨的同时，在很大程度上缩短了患者整体治疗的时间使患者更容易接受。术后复发率尚可，没有遗留明显的并发症。
 

ive
1.identifing the gene variation associated with non-syndromic cleft lip and palate in a chinese family using linked-read technology.
2.1 to investigate horizontal maxillary osteotomy stability after using bone grafts for the treatment of patients with unilateral cleft lip and palate.
2.2 to present the clinical results and complications of a two-stage surgical approach using a modified external distraction system that consists of maxillary distraction and then maxillary fixation.
methods
1.in this study, we analyzed a non-syndromic cleft lip and palate multiplex family, collected the epidemiological information, and mapped the pedigree. the genomic dna were obtained from whole blood sample. linked-read sequencing was performed to achieve genome-wide variants. based on hg38, long ranger software was used to map the variants, annovar was used to annotate the variants. the candidate genes variants were selected according to the location of the variants, allele frequencies, function, conservation, disease risk, as well as their association with craniofacial development and cleft lip and palate.
2.1 fifty-eight patients with maxillary hypoplasia were included, who required a maxillary advancement of 6 to 9 mm. the test group was comprised of 28 patients who underwent mandibular outer cortex bone grafting in the gaps created by a modified le fort i osteotomy. the control group was comprised of 30 patients who underwent a traditional le fort i osteotomy without bone grafts.
2.2 we treated eleven patients with cleft lip and palate by two-stage maxillary distraction osteogenesis. lateral cephalograms taken before the first operation, after distraction, two weeks after the second operation, and one year after treatment were used to examine maxillofacial morphology. five patients were treated with transverse expansion at the same time. computer tomography was taken before and after consolidation to analyze the effect of maxillary expansion. velopharyngeal function was evaluated by a speech therapist.
results
1. we identified two candidate variants for this non-syndromic cleft lip and palate multiplex family including a missense of axin2(c.2018g＞a, p.s673f) and a deletion of 155 kilo-base region overlapping adam3a and adam5 (chr8, 39374555-39529710) .
2.1 in the test group, the mean maxillary horizontal advancement was 7.13±0.7mm (range: 6.01–8.23 mm), and the mean postoperative horizontal relapse was 25.07±6.64%. in the cg, the mean maxillary horizontal advancement was 6.90±0.55mm (range: 6.05–7.39 mm), and the mean postoperative horizontal relapse was 24.89±4.25%. there were no significant betweengroup differences in the mean horizontal relapses.
2.2 the mean movements of the maxilla forwards and downwards at point a were 13.9mm and 7.7mm at the completion of distraction and those at point b were 5.3mm backwards and 8.5mm downwards. clockwise rotation was observed in maxilla and mandible. mouth opening was limited at this time. after the maxilla was fixed, compared with pre-operation, point a moved 10.1mm anteriorly and 3.4mm inferiorly and point b moved 2.4mm posteriorly and 4.7mm inferiorly. limitation of mouth opening was relieved after maxillary fixation. the mean relapse of point a one year postoperatively was 22.2% horizontally and 51.6% vertically. after expansion in 5 patients, the width of dental arch was significantly increased. the mean width between cusps of canines increased 6.4mm, and between mesial-buccal cusps of the first molars increased 6.2mm. velopharyngeal function was unchanged by the operation.
conclusion
1. the two gene variants may be involved in the etiology of oral clefts.
2.1 the use of mandibular outer cortex bone grafts in patients with unilateral cleft lip and palate did not increase postoperative stability when the maxillary advancement was 6 to 9 mm.
2.2 we conclude that the method had advantages that included the short duration of wearing distractors and increased acceptance by patients. the modified external device advanced the midface sufficiently.