A novel missense mutation of RPGR identified from retinitis pigmentosa affects splicing of the ORF15 region and causes loss of transcript heterogeneity.

Mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene, are the major cause of X-linked retinitis pigmentosa (RP), in which exon open reading frame 15 (ORF15) of RPGR has been implicated to play a substantial role. We identified a novel hemizygous missense mutation E585K of RPGR from whole-exome sequencing of RP. RNA-Seq analysis and functional study were conducted to investigate the underlying pathogenic mechanism of the mutation. Our results showed that the mutation actually affected RPGR ORF15 splicing. RNA-Seq analysis of the human retina followed by validation in cells revealed a complex splicing pattern near the 3' boundary of RPGR exon 14 in the ORF15 region, resulting from a variety of alternative splicing events (ASEs). The wildtype RPGR mini-gene expressed in human 293T cells confirmed these ASEs in vitro. In contrast, without new RNA species detected, the mutant mini-gene disrupted the splicing pattern of the ORF15 region, and caused loss of RPGR transcript heterogeneity. The RNA species derived from the mutant mini-gene were predominated by a minor out-of-frame transcript that was also observed in wildtype RPGR, resulting from an upstream alternative 5' splice site in exon 14. Our findings therefore provide insights into the influence of RPGR exonic mutations on alternative splicing of the ORF15 region, and the underlying molecular mechanism of RP.

A female patient carrying a novel DMD mutation with non-random X-chromosome inactivation from a DMD family.

OBJECTIVE: To analyze the clinical phenotype and genetic characteristics of a female proband carrying a novel mutation in the DMD gene with non-random X-chromosome inactivation in a large pedigree with pseudohypertrophic muscular dystrophy. METHODS: Clinical information of the female proband, her monozygotic twin sister, and other family members were collected. Potential pathogenic variants were detected with Multiplex Ligation-dependent Probe Amplification (MLPA) and whole-exome sequencing (WES). Methylation-sensitive restriction enzyme (HhaI) was employed for X-chromosome inactivation analysis. RESULTS: The proband was a female over 5 years old, displayed clinical manifestations such as elevated creatine kinase (CK) levels and mild calf muscle hypertrophy. Her monozygotic twin sister exhibited normal CK levels and motor ability. Her uncle and cousin had a history of DMD. WES revealed that the proband carried a novel variant in the DMD (OMIM: 300,377) gene: NM_004006.3: c.3051_3053dup; NP_003997.2: p.Tyr1018*. In this pedigree, five out of six female members were carriers of this variant, while the cousin and uncle were hemizygous for this variant. X-chromosome inactivation analysis suggested non-random inactivation in the proband. CONCLUSION: The c.3051_3053dup (p.Tyr1018*) variant in the DMD gene is considered to be the pathogenic variant significantly associated with the clinical phenotype of the proband, her cousin, and her uncle within this family. Integrating genetic testing with clinical phenotype assessment can be a valuable tool for physicians in the diagnosis of progressive muscular dystrophies, such as Becker muscular dystrophy (BMD) and Duchenne muscular dystrophy (DMD).

Engineered CBEs based on Macaca fascicularis A3A with improved properties for precise genome editing.

Cytidine deaminase defines the properties of cytosine base editors (CBEs) for C-to-T conversion. Replacing the cytidine deaminase rat APOBEC1 (rA1) in CBEs with a human APOBEC3A (hA3A) improves CBE properties. However, the potential CBE application of macaque A3A orthologs remains undetermined. Our current study develops and evaluates engineered CBEs based on Macaca fascicularis A3A (mA3A). Here, we demonstrate that BE4-mA3A and its RNA-editing-derived variants exhibit improved CBE properties, except for DNA off-target activity, compared to BE3-rA1 and BE4-rA1. Unexpectedly, deleting Ser-Val-Arg (SVR) in BE4-mA3A dramatically reduces DNA and RNA off-target activities and improves editing accuracy, with on-target efficiency unaffected. In contrast, a chimeric BE4-hA3A-SVR+ shows editing efficiency increased by about 50%, with other properties unaffected. Our findings demonstrate that mA3A-based CBEs could provide prototype options with advantages over rA1- and hA3A-based CBEs for further optimization, highlighting the importance of the SVR motif in defining CBE intrinsic properties.

Comparative Analysis of Molecular Landscape in Mouse Models and Patients Reveals Conserved Inflammation Pathways in Age-Related Macular Degeneration.

Purpose: The purpose of this study was to identify key genes and their regulatory networks that are conserved in mouse models of age-related macular degeneration (AMD) and human AMD. Methods: Retinal RNA-Seq was performed in laser-induced choroidal neovascularization (CNV) mice at day 3 and day 7 after photocoagulation. Mass spectrometry-based proteomic analysis was performed with retinas collected at day 3. Retinal RNA-Seq data was further compared among mouse models of laser-induced CNV and NaIO3-induced retinal degeneration (RD) and a large AMD cohort. Results: Retinal RNA-Seq revealed upregulated genes and pathways related to innate immunity and inflammation in mice with CNV, with more profound changes at the early stage (day 3). Proteomic analysis further validated these differentially expressed genes and their networks in retinal inflammation during CNV. Notably, the most evident overlap in the retina of mice with laser-induced CNV and NaIO3-induced RD was the upregulation of inflammation-related genes, pointing to a common vital role of retinal inflammation in the early stage for both mouse AMD models. Further comparative transcriptomic analysis of the mouse AMD models and human AMD identified 48 conserved genes mainly involved in inflammation response. Among them, B2M, C3, and SERPING1 were upregulated in all stages of human AMD and the mouse AMD models compared to controls. Conclusions: Our study demonstrates conserved molecular changes related to retinal inflammation in mouse AMD models and human AMD and provides new insight into the translational application of these mouse models in studying AMD mechanisms and treatments.

Application of long read sequencing in rare diseases: The longer, the better?

Rare diseases encompass a diverse group of genetic disorders that affect a small proportion of the population. Identifying the underlying genetic causes of these conditions presents significant challenges due to their genetic heterogeneity and complexity. Conventional short-read sequencing (SRS) techniques have been widely used in diagnosing and investigating of rare diseases, with limitations due to the nature of short-read lengths. In recent years, long read sequencing (LRS) technologies have emerged as a valuable tool in overcoming these limitations. This minireview provides a concise overview of the applications of LRS in rare disease research and diagnosis, including the identification of disease-causing tandem repeat expansions, structural variations, and comprehensive analysis of pathogenic variants with LRS.

Pretreatment with Bifidobacterium longum BAA2573 ameliorates dextran sulfate sodium (DSS)-induced colitis by modulating gut microbiota.

Objectives: Inflammatory bowel disease (IBD) is a chronic lifelong inflammatory disease. Probiotics such as Bifidobacterium longum are considered to be beneficial to the recovery of intestinal inflammation by interaction with gut microbiota. Our goals were to define the effect of the exclusive use of BAA2573 on dextran sulfate sodium (DSS)-induced colitis, including improvement of symptoms, alleviation of histopathological damage, and modulation of gut microbiota. Methods: In the present study, we pretreated C57BL/6J mice with Bifidobacterium longum BAA2573, one of the main components in an over-the-counter (OTC) probiotic mixture BIFOTO capsule, before modeling with DSS. 16S rDNA sequencing and liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based non-targeted metabolomic profiling were performed with the collected feces. Results: We found that pretreatment of Bifidobacterium longum BAA2573 given by gavage significantly improved symptoms and histopathological damage in DSS-induced colitis mice. After the BAA2573 intervention, 57 genera and 39 metabolites were significantly altered. Pathway enrichment analysis demonstrated that starch and sucrose metabolism, vitamin B6 metabolism, and sphingolipid metabolism may contribute to ameliorating colitis. Moreover, we revealed that the gut microbiome and metabolites were interrelated in the BAA2573 intervention group, while Alistipes was the core genus. Conclusion: Our study demonstrates the impact of BAA2573 on the gut microbiota and reveals a possible novel adjuvant therapy for IBD patients.

Host A-to-I RNA editing signatures in intracellular bacterial and single-strand RNA viral infections.

Background: Microbial infection is accompanied by remodeling of the host transcriptome. Involvement of A-to-I RNA editing has been reported during viral infection but remains to be elucidated during intracellular bacterial infections. Results: Herein we analyzed A-to-I RNA editing during intracellular bacterial infections based on 18 RNA-Seq datasets of 210 mouse samples involving 7 tissue types and 8 intracellular bacterial pathogens (IBPs), and identified a consensus signature of RNA editing for IBP infections, mainly involving neutrophil-mediated innate immunity and lipid metabolism. Further comparison of host RNA editing patterns revealed remarkable similarities between pneumonia caused by IBPs and single-strand RNA (ssRNA) viruses, such as altered editing enzyme expression, editing site numbers, and levels. In addition, functional enrichment analysis of genes with RNA editing highlighted that the Rab GTPase family played a common and vital role in the host immune response to IBP and ssRNA viral infections, which was indicated by the consistent up-regulated RNA editing of Ras-related protein Rab27a. Nevertheless, dramatic differences between IBP and viral infections were also observed, and clearly distinguished the two types of intracellular infections. Conclusion: Our study showed transcriptome-wide host A-to-I RNA editing alteration during IBP and ssRNA viral infections. By identifying and comparing consensus signatures of host A-to-I RNA editing, our analysis implicates the importance of host A-to-I RNA editing during these infections and provides new insights into the diagnosis and treatment of infectious diseases.

[The effect of orthognathic surgery on speech function in patients with skeletal Class â ¢ malocclusion].

PURPOSE: To investigate the influence of the position of the upper and lower jaws on the anatomical structure of pharynx before and after orthognathic surgery in patients with skeletal Class Ⅲ malocclusion. METHODS: Craniofacial CT scan and speech data were collected from 31 patients with skeletal Class Ⅲ malocclusion before and 3 months after surgery. The collected CT data was imported into Dolphin imaging 11.95 software to establish a digital original model, and the anatomical structure of the pharynx was measured and analyzed. Speech data were analyzed objectively and subjectively by Computerized Speech Lab 4500b and professional speech specialists. Statistical analysis was performed using SPSS 24.0 software package. RESULTS: The distance from the lower edge of the soft palate to the posterior pharyngeal wall, the shortest distance from the posterior margin of the tongue to the posterior pharyngeal wall and its corresponding cross-sectional area were significantly different from those before surgery (P<0.05). The changes of SNA, SNB, ANB, OJ, and OBJ before and after surgery were significant in this series. Importantly, the speech intelligibility of orthognathic patients before and after surgery changed significantly subjectively (P<0.05). Objectively, the postoperative vowels /a/B2, B3, B4, /i/B1,B2, /u/B1,B2 and B4 of the patients were significantly different from those before surgery. There was no significant difference in the lower limit frequency of the consonants /x/, /zh/, /s/, the energy value of /zh/ and the grammatical form of /z/ before and after surgery. The maxillary advancement distance was highly correlated or significantly correlated with △S1, △VOP, and voice changes. CONCLUSIONS: Orthognathic surgery moves the upper and lower jaws to cause changes in the anatomy of the pharyngeal cavity, leading to changes of postoperative speech.

Cytotoxic T-Cell Trafficking Chemokine Profiles Correlate With Defined Mucosal Microbial Communities in Colorectal Cancer.

The involvement of gut microbiota in T-cell trafficking into tumor tissue of colorectal cancer (CRC) remains to be further elucidated. The current study aimed to evaluate the expression of major cytotoxic T-cell trafficking chemokines (CTTCs) and chemokine-associated microbiota profiles in both tumor and adjacent normal tissues during CRC progression. We analyzed the expression of chemokine C-X-C motif ligands 9, 10, and 11 (CXCL9, CXCL10, and CXCL11), and C-C motif ligand 5 (CCL5), characterized gut mucosa-associated microbiota (MAM), and investigated their correlations in CRC patients. Our results showed that the expression of CXCL9, CXCL10, and CXCL11 was significantly higher in tumor than in adjacent normal tissues in 136 CRC patients. Notably, the high expression of CXCL9 in tumor tissues was associated with enhanced CD8+ T-cell infiltration and improved survival. Moreover, the MAM in tumor tissues showed reduction of microbial diversity and increase of oral bacteria. Microbial network analysis identified differences in microbial composition and structure between tumor and adjacent normal tissues. In addition, stronger associations between oral bacteria and other gut microbes were observed. Furthermore, the correlation analysis between the defined MAM and individual CTTCs showed that the CTTCs' correlated operational taxonomic units (OTUs) in tumor and adjacent normal tissues rarely overlap with each other. Notably, all the enriched OTUs were positively correlated with the CTTCs in either tumor or adjacent normal tissues. Our findings demonstrated stronger interactions between oral bacteria and gut microbes, and a shifted correlation pattern between MAM and major CTTCs in tumor tissues, underlining possible mechanisms of gut microbiota-host interaction in CRC.

The influence of sample size and gender composition on the meta-analysis conclusion of platelet-rich plasma treatment for osteoarthritis.

OBJECTIVE: The magnitude of the therapeutic effects of intra-articular injection of platelet-rich plasma (PRP) on osteoarthritis (OA) is still under debate. The goal of this study that was a systematic review of randomised controlled trials â€‹of PRP injections for the treatment of OA was to elucidate the therapeutic efficacy of PRP. METHODS: Electronic databases of PubMed, CENTRAL, EMBASE, EBSCO, ClinicalTrials.gov, and International Clinical Trials Registry Platform â€‹were searched from inception to June 2018 for RCTs that compared PRP injections to controls in patients with OA. A random-effects approach was used to compile data and subgroups according to trial size (large trials versus small trials), patient profile (age and gender), and PRP preparation method was performed. RESULTS: Thirty trials met the inclusion criteria and were analysed. All results had unexplained statistical heterogeneity. Patients treated with PRP compared with control showed statistically relevant pain relief and function improvement at short term (standardised mean difference [SMD] â€‹= â€‹-0.62, 95% confidence interval [CI]: -0.98 to -0.27, P â€‹= â€‹0.0006, SMD â€‹= â€‹-0.74, 95% CI: -1.11 to 0.36, P â€‹= â€‹0.0001, respectively), medium term (SMD â€‹= â€‹-0.53, 95% CI: -0.83 to -0.23, P â€‹= â€‹0.0006, SMD â€‹= â€‹-0.50, 95% CI: -0.75 to -0.25, P â€‹= â€‹0.0006), and long term (SMD â€‹= â€‹-0.69, 95% CI: -1.08 to -0.30, P â€‹= â€‹0.0006, SMD â€‹= â€‹-0.68, 95% CI: -0.1.09 to -0.27, P â€‹= â€‹0.001, respectively). A subgroup analysis of the data from large trials and from trials composed of less than 50% female patients revealed that therapeutic effects of the treatment are insignificant. CONCLUSIONS: According to the currently available data, PRP injections are beneficial for pain relief and function improvement in patients with OA. This meta-analysis, however, demonstrated that the efficacy of PRP is related to sample size and gender composition. Thus, more randomised controlled trials of high quality and larger patient size, also including gender aspects, are required to understand this phenomenon. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The translation potential of this meta-analysis is that provided another perspective to analyse the treatment effect of PRP for OA. In future research, phenotypes subpopulation and gender difference of OA patient should be considered for PRP treatment.

[Identification of a novel TP53 germline mutation in one child with adrenocortical carcinoma].

OBJECTIVE: To detect the germline TP53 gene mutation in a child with pediatric adrenocortical carcinoma (ADCC) in order to provide genetic diagnosis and counseling. METHODS: Genomic DNA was extracted from peripheral blood from a girl with ADCC and her parents. All TP53 exons and their flanking intronic sequences were PCR-amplified and subjected to automatic DNA sequencing. RESULTS: Direct sequencing of PCR products revealed a heterozygous G insertion between nucleotide 522 and 523 (c.522-523insG) in TP53 exon 5. This novel mutation is predicted to result in a frame shift at codon 175, producing a new reading frame ending in a stop at position 6 (p.R175AfsX6). The same heterozygous mutation was also found in her father, but not in her mother. CONCLUSION: A novel germline mutation in the TP53 gene has been identified in one case with pediatric ADCC.

The relationship between maternal corticosteroid use and orofacial clefts-a meta-analysis.

The aim was to evaluate the relationship between maternal corticosteroid use during first trimester of pregnancy and risk of orofacial clefts (OC). The overall findings showed a certain association between maternal corticosteroid use and occurrence of OC, compared with non-users (OR=1.16 [95% CI: 1.01-1.33]). When study type was considered this association was significant only for case-control studies (OR=1.22 [95% CI: 1.02-1.47]), and not for cohort studies (OR=1.09 [95% CI: 0.88-1.34]) when there are many confounders (dose, route of application, disease etc.) and biases (re-call, loss-to follow-up etc.) that still need to be considered. A subgroup analysis based on the type of OC gave an overall OR of 1.41 (95% CI: 1.14-1.74) in the case-control studies for cleft lip with or without palate (CL/P) and 1.09 (95% CI: 0.80-1.48) for cleft palate only (CPO), when comparing maternal corticosteroid users with non-users. However, for cohort studies, the overall OR for CL/P is 1.06 (95% CI: 0.82-1.37) and 1.20 (95% CI: 0.83-1.75) for CPO. The absolute risk of facial cleft after prenatal exposure to corticosteroids, if any, is small.

An effective seed protection method for planting Zostera marina (eelgrass) seeds: Implications for their large-scale restoration.

We describe an innovative method of planting Zostera marina (eelgrass) seeds in which hessian bags filled with high-silted sediments are used as a seed protecting device. Here, we evaluated the effectiveness of the method through a field seed-sowing experiment over a three year period. The suitable seed planting density required by the seeds of Z. marina in this method was also investigated. In the spring following seed distribution, seedling establishment rate of Z. marina subjected to different seed densities of 200-500seedsbag(-1) ranged from 16% to 26%. New eelgrass patches from seed were fully developed and well maintained after 2-3years following distribution. The seed planting density of 400seedsbag(-1) may be the most suitable for the establishment of new eelgrass patches. Our results demonstrate that seed-based restoration can be an effective restoration tool and the technique presented should be considered for future large-scale Z. marina restoration projects.

1,25(OH)2D3 inhibits the deleterious effects induced by high glucose on osteoblasts through undercarboxylated osteocalcin and insulin signaling.

Diabetes mellitus (DM) is associated with multiple skeletal disorders, and vitamin D may play a functional role in the preservation of glucose tolerance. However, the relationship between vitamin D deficiency and DM is not well known. The aim of this study was to investigate the potential molecular link between 1,25(OH)(2)D(3) regulation and glucose homeostasis. Rat primary osteoblasts were cultured in different conditioned medium: normal glucose, high glucose, high glucose and insulin, high glucose and 1,25(OH)(2)D(3), high glucose and insulin and 1,25(OH)(2)D(3). The activity of osteoblasts was measured by cell viability, alkaline phosphatase and osteocalcin assay. The potential mechanism of how 1,25(OH)(2)D(3) affect insulin sensitivity was investigated by the assay of insulin receptor (IR) and vitamin D receptor (VDR) expression, and undercarboxylated osteocalcin (ucOC) level. The combined treatment has the strongest effect of inhibiting the deleterious effects induced by high glucose on osteoblasts, and it promoted the %ucOC value to approximately 40%, which is much higher than that in high glucose without treatment. Levels of IR and VDR of osteoblasts in combined treatment culture increased significantly compared with that in high glucose without treatment. So maybe 1,25(OH)(2)D(3) promotes insulin sensitivity of osteoblasts by activating insulin signaling and simultaneously stimulating ucOC secretion, which in turn regulate insulin production and sensitivity. 1,25(OH)(2)D(3) might be beneficial not only for diabetes, but also, for osteoporosis by promoting bone formation.

[Distraction osteogenesis for correction of cleft palate in rhesus-histological and fluorescent labeling study].

OBJECTIVE: To investigate the mechanism of new bone formation in the distraction osteogenesis (DO) for correction of cleft palate (CP) in rhesus. METHODS: CP was created by operation in 23 rhesus. The CP was corrected with DO in 21 animals as experimental group. The distraction rate was 0.8 mm per day, two times a day. The bone fragments were fixed after cleft closure, every 3 animals were sacrificed to get specimen after 1, 2, 4, 6, 8, 12, 24 weeks of fixation. 6 days before sacrifice, tetracycline was administrated for labeling (30 mg/kg). RESULTS: The hard and soft tissue def of fixation. At the same time, the bone volume and calcification between the distraction gap increased. The cleft in the control group could not b ect was successfully closed with DO by intramembrane osteogenesis. The new formed bone was remodeling and became maturation during the period e corrected spontaneously. CONCLUSIONS: The DO can successfully correct both the soft and hard tissue defect in CP by intramembrane osteogenesis. The fixation is important for remodeling and maturation of the new formed bone.

[Ultrastructural and element spectrometric analysis of distraction osteogenesis for reconstruction of cleft palate in rhesus macaque model].

OBJECTIVE: To study the ultrastructure and Ca/P element spectrometry of distraction osteogenesis (DO) for reconstruction of cleft palate (CP), so as to explore the osteogenesis and remodeling of new bone in situ. METHODS: 23 rhesus macaques were operated to establish animal models of CP. 2 monkeys didn't received DO as controls. The other 21 monkeys in experimental group underwent DO to correct both bony and soft tissue defects in palate. The distraction was performed at a rate of 0.8 mm/d, twice a day until the cleft was closed. After fixation for 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were sacrificed to get the specimens at the distraction gap. The scanning electron microscopic study and Ca, P elements spectrometric analysis were adopted. There were also two unoperated animals as sham group. RESULTS: After fixation for 1-2 weeks, the distraction gap was full of collagen fibers oriented along vector of distraction. Few trabeculae was seen at the margin area. After fixation for 4-6 weeks, active osteogenesis was presented with new formed bone trabeculae and abundant cellular component. After fixation for 8-12 weeks, the new formed bone became mature and couldn't distinguish from the normal bone. 24 weeks later, the bone between the distraction gap had a similar structure to the normal bone. Elements spectrometric analysis results indicated that in early stage of osteogenesis, the P and S peaks were relatively high while the Ca peak was much lower. During the late stage, the S peak was obviously decreased, and Ca/P ratio increased to normal level as in the empty control group. CONCLUSIONS: The CP can be corrected by DO. The new bone between the distraction gap is formed and remodeled through intramembraneous osteogenesis.

[Bone morphogenetic protein expression in distraction osteogenesis correction for cleft palate in Rhesus monkeys].

OBJECTIVE: To study the expression and distribution of bone morphogenetic protein (BMP) in newly formed bone by distraction osteogenesis (DO), and to explore the mechanism of the DO bone formation and remodeling. METHODS: The cleft palate (CP) experimental animal models (23 Rhesus monkeys) were established surgically. In experimental group (21 Rhesus monkeys), the palatal defects were corrected by means of DO at the rhythm of 0.4 mm twice per day. The specimens were retrieved under euthanasia at 1, 2, 4, 6, 8, 12, 24 weeks intervals respectively in retention period. BMP immunohistochemical study was then performed. The blank control and experimental group (each of 2 animals) were set for comparison study. RESULTS: The immunohistochemical study showed that BMP existed mainly in cytoplasma of osteoblasts, during the process of new bone formation. In early stage of 1 or 2 weeks, abundant osteoblasts aggregating on surfaces of the new bone trabeculae with positive DAB dye were observed. Through 4 to 6 weeks, the proliferative osteoblasts with very strong positive DAB dye indicating BMP expression were recorded. From 8 to 12 weeks, the expression of BMP and quantity of osteoblasts decreased gradually while more matured new bone structures were observed. CONCLUSION: During the whole retention period, the expression of BMP showed a tendency from weak to strong and then to final cessation, this indicated a process of formation, remodeling and maturation of osteogenesis.

[Expression of osteopontin and osteocalcin during distraction osteogenesis on rhesus with cleft palate].

OBJECTIVE: To study the mechanism of new bone formation and remodeling of distraction osteogenesis (DO) by analysis of the expression of osteopontin (OPN) and osteocalcin (OC). METHODS: Rhesus were operated to reconstruct the animal model of cleft palate (CP). The CP was closed by DO in experimental group(n = 21). After consolidation of 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were killed to collect the specimens, respectively. The OPN and OC and their mRNA were detected quantitatively by Real-time RT-PCR and ELISA, respectively. The animals in control group (n = 2) and sham group (n = 2) were used as control. RESULTS: The mRNA expression of OPN increased since 2nd week of consolidation and reached the peak at 4th week (7.59 +/- 0.37). The mRNA expression of OC was up-regulated since 4th week, and reach the peak at 6th week (7.94 +/- 0.31). Then they decreased to about the level in sham group at 24th week (P > 0.05). The OPN and OC were highly expressed during 4 to 6 weeks of consolidation. During 8 to 12 weeks, they decreased like their mRNA expression. CONCLUSION: The intramembraneous new bone formation after DO can reconstruct the bone defect of CP. The new formed bone can be remodeled to be quite normal bone tissue.

[Expression of insulin-like growth factor-1 and alkaline phosphatase in the reconstruction of cleft palate with distraction osteogenesis in rhesus].

OBJECTIVE: To investigate the osteogenesis mechanism by analysis of the expression of insulin-like growth factor-I (IGF-1) and alkaline phosphatase (ALP) in the reconstruction of cleft palate (CP) with distraction osteogenesis (DO) in rhesus. METHODS: The CP animal models were established surgically. 21 rhesus in experimental group underwent DO to close the soft and bony defect, followed by consolidations. Every 3 animals were killed and the specimen were taken out after consolidation of 1, 2, 4, 6, 8, 12, 24 weeks. The mRNA of IGF-1 and ALP were detected with Real-time RT-PCR technique. The expression of IGF-1 and ALP was quantitatively analyzed by ELISA. The results were compared with those in control and sham groups (each of 2 animals), respectively. RESULTS: Since consolidation, the mRNA of IGF-1 and ALP increased significantly at one week and reached the peak at two weeks, but decrease to control level after 12 weeks of consolidation. The expression of IGF-1 also increased to peak level after two weeks of consolidation. The expression of ALT increased significantly since consolidation and reach the peak value after six weeks. They all decreased to nearly control level after 8-12 weeks. CONCLUSIONS: The palate cleft can be successfully closed with new formed bone after DO. The mechanism of bone consolidation is intramembranous bone formation.