Deletion of cis-regulatory Element in FOXL2 Promoter in a Chinese Family of Type II Blepharophimosis-ptosis-epicanthus Inversus Syndrome with Polydactyly.

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a relatively uncommon autosomal-dominant genetic disorder, primarily attributed to mutations in the forkhead box L2 (FOXL2) gene. Albeit the involvement of protein-coding regions of FOXL2 has been observed in the majority of BPES cases, whether deficiencies in regulatory elements lead to the pathogenesis remains poorly understood. Herein, an autosomal-dominant BPES type II family was included. Peripheral venous blood has been collected, and genomic DNA has been extracted from leukocytes. A whole exome sequencing analysis has been performed and analyzed (Deposited in NODE database: OER422653). The promoter region of FOXL2 was amplified using polymerase chain reaction (PCR). The luciferase reporter assay was performed to identify the activity of this region. In this study, we present a Chinese family diagnosed with type II BPES, characterized by the presence of small palpebral fissures, ptosis, telecanthus, and epicanthus inversus. Notably, all male individuals within the family display polydactyly. A 225-bp deletion in the 556-bp 5'-upstream to transcription start site of FOXL2 , decorated by multiple histone modifications, was identified in affected members of the family. This deletion significantly decreased FOXL2 promoter activity, as measured by the luciferase assay. Conclusively, a novel 255-bp-deletion of the FOXL2 promoter was identified in Chinese families with BPES. Our results expand the spectrum of known FOXL2 mutations and provide additional insight into the genotype-phenotype relationships of the BPES pathogenesis. In addition, this study indicates the important role of genetic screening of cis-regulatory elements in testing heritable diseases.

A non-invasive smart scaffold for bone repair and monitoring.

Existing strategies for bone defect repair are difficult to monitor. Smart scaffold materials that can quantify the efficiency of new bone formation are important for bone regeneration and monitoring. Carbon nanotubes (CNT) have promising bioactivity and electrical conductivity. In this study, a noninvasive and intelligent monitoring scaffold was prepared for bone regeneration and monitoring by integrating carboxylated CNT into chemically cross-linked carboxymethyl chitosan hydrogel. CNT scaffold (0.5% w/v) demonstrated improved mechanical properties with good biocompatibility and electrochemical responsiveness. Cyclic voltammetry and electrochemical impedance spectroscopy of CNT scaffold responded sensitively to seed cell differentiation degree in both cellular and animal levels. Interestingly, the CNT scaffold could make up the easy deactivation shortfall of bone morphogenetic protein 2 by sustainably enhancing stem cell osteogenic differentiation and new bone tissue formation through CNT roles. This research provides new ideas for the development of noninvasive and electrochemically responsive bioactive scaffolds, marking an important step in the development of intelligent tissue engineering.

Orbital Growth is Associated with Eyeball Size: A Study Using CT-based Three-dimensional Techniques.

AIM OF THE STUDY: This study was aimed to investigate the growth patterns and the relationship of the eyeball and the orbit using computed tomography (CT)-based three-dimensional (3D) techniques. MATERIALS AND METHODS: A total of 175 Chinese patients who had undergone craniofacial or orbital CT scans were enrolled. This study only included data from the unaffected eye and orbit. Images were processed using 3D reconstruction to obtain the eyeball and the orbit parameters. RESULTS: In early postnatal years, the sizes of eyeball and orbit increased significantly with age (p < 0.001) and reached a turning point at a critical age (8.967 and 12.800 years for the eyeball and orbit volume, respectively). The orbital index and orbital depth index, showing the shape of the orbital aperture and walls, decreased significantly with age (p < 0.001). In all ages, the orbit size was correlated with eyeball size (p < 0.001). The eye-orbit index, equivalent to the ratio of eye volume to orbital volume, declined steadily with age (p < 0.001). CONCLUSIONS: The eyeball and orbit developed rapidly in early postnatal years, and then matured at a critical age. The eyeball size significantly contributed to the orbital growth; this contribution may be reduced as the eye-orbit index decreased with age. To the best of our knowledge, this is the first report on the growth and interrelation of the eyeball and the orbit using CT-based 3D techniques.

Targeting Local Osteogenic and Ancillary Cells by Mechanobiologically Optimized Magnesium Scaffolds for Orbital Bone Reconstruction in Canines.

Large-sized orbital bone defects have serious consequences that destroy orbital integrity and result in maxillofacial deformities and vision loss. The treatment of orbital bone defects is currently palliative and not reparative, suggesting an urgent demand for biomaterials that regenerate orbital bones. In this study, via alloying, extrusion and surface modification, we developed mechanobiologically optimized magnesium (Mg) scaffolds (Ca-P-coated Mg-Zn-Gd scaffolds, referred to as Ca-P-Mg) for the orthotopic reconstruction of large-sized orbital bone defects. At 6 months after transplanting the scaffolds to a clinically relevant canine large animal model, large-sized defects were successfully bridged by an abundance of new bone with normal mechanical properties that corresponded to gradual degradation of the implants. The osteogenic and ancillary cells, including vascular endothelial cells and trigeminal neurons, played important roles in this process. The scaffolds robustly enhanced bone marrow mesenchymal stem cell (BMSC) osteogenic differentiation. In addition, the increased angiogenesis including increased ratio of the specific endothelial subtype CD31hi endomucinhi (CD31hiEmcnhi) endothelial cells can facilitate osteogenesis. Furthermore, the scaffolds trigger trigeminal neurons via transient receptor potential vanilloid subtype 1 (Trpv1) to produce the neuropeptide calcitonin gene-related peptide (CGRP), which promotes angiogenesis and osteogenesis. Overall, our investigations revealed the efficacy of Ca-P-Mg scaffolds in healing orbital bone defects and warrant further exploration of these scaffolds for clinical applications.

Prediction of late displacement of the globe in orbital blowout fractures.

PURPOSE: To establish a linear measuring method in computed tomographic (CT) images to predict the displacement of the globe late after orbital blowout fracture. METHODS: Subjects were retrospectively included. Inclusion criteria were as follows: (1) adult subjects (≥18 years old at the time of trauma); (2) unilateral orbital medial-wall and/or floor fractures; (3) CT examination at least 30 days after trauma. Exclusion criteria were as follows: (1) facial or orbital fracture extending to other parts of the orbit than medial-wall and/or floor; (2) history of orbital or ocular abnormality other than the orbital trauma; (3) severe ocular trauma accompanied by the orbital trauma; (4) orbital fracture treated surgically before the CT examination. A co-ordinate system was built based on the orbital CT scans. Displacements of orbital walls, displacement of the globe and relative location of the fracture site were measured. Correlations between the variables were investigated. RESULTS: Ninety-nine per cent of fracture sites of the medial wall and 100% of fracture sites of the floor were posterior to the centre of the unaffected globe. The affected globe moved significantly medially (p < 0.001) and backwards (p < 0.001) in pure medial-wall fracture; backwards (p < 0.001) and downwards (p = 0.017) in pure floor fracture; and medially (p < 0.001), backwards (p < 0.001) and downwards (p < 0.001) in medial-wall and floor fractures. Displacement of the globe was correlated with displacements of the orbital walls, and the regression formulae were therefore fitted. Application of the formulae revealed that the same extent of orbital wall displacement caused more displacement of the globe in female patients than in male patients. CONCLUSIONS: A linear measuring method in a three-dimensional co-ordinate system was established to identify the displacements of orbital walls and the displacement of the globe in orbital blowout fractures. The regression formulae generated in this study might be used in clinical practice to predict late displacement of the globe by measuring the displacements of orbital walls.

Measurement of Intra-Orbital Structures in Normal Chinese Adults Based on a Three-Dimensional Coordinate System.

PURPOSE OF THE STUDY: This study was to establish a three-dimensional (3D) coordinate system and to study the normal dimensions of intra-orbital structures in Chinese adults. MATERIALS AND METHODS: One hundred and forty-five adult Chinese were selected from patients who had undergone cranio-facial computed tomography scans with diagnosis other than orbital or ocular abnormality. An orbital 3D coordinate system was built on the basis of the scans. Morphological variables of intra-orbital structures were measured in this coordinate system. Bilateral symmetry, sexual dimorphism, and correlations between variables were investigated. RESULTS: No evident laterality was found in bilateral intra-orbital structures. The distance from the center of the eyeball to the prechiasmatic groove, the length of the optic nerve, and the thickness of rectus extraocular muscles were larger in males than in females. No sex-related difference was observed in the anteroposterior diameter of the eyeball or the exophthalmometric value. The exophthalmometric value was found to be related to the anteroposterior diameter of the eyeball, whereas the y-coordinate of the center of the eyeball had no correlation with the anteroposterior diameter of the eyeball. The optic nerve length was closely correlated to the distance from the center of the eyeball to the prechiasmatic groove. CONCLUSIONS: The 3D coordinate system and measurement method established in this study can be applied to the standardization of orbital morphometry. The measurements obtained from normal Chinese adults may provide reference values for the morphology of intra-orbital structures.

Epidemiologic characteristics and outcomes of open globe injury in Shanghai.

AIM: To investigate the epidemiologic characteristics and outcomes of open globe injury in Shanghai. METHODS: A retrospective study was conducted for 148 unilateral open globe injury cases presenting to a tertiary referral hospital of Shanghai. Electronic medical records were reviewed and phone surveys were conducted to collect and analyze 1) background of patient; 2) setting of injury and clinical signs at presentation; 3) treatment procedure and outcome; 4) quality of life after injury. RESULTS: There were more male patients (77.03%) than females (22.97%), more temporary habitants (79.05%) than residents (20.95%). The subjects in this study presented a significantly lower constitutional status of education than that of the whole Shanghai population (P<0.001). Occupational injury was the first cause of injuries (39.86%), followed by home accident (20.27%), road accident (16.89%), violent behavior (16.89%) and outdoor injury (6.08%). The 143 subjects (96.62%) were not wearing spectacles at the time of injury. Of all patients, 77 subjects (52.03%) had the outcome of no vision (including enucleation). The classification and regression tree (CART) prognosis presents 59.58% sensitivity to predict visual survival correctly and 80.19% specificity to predict no vision correctly. The patients whose injured eye had no vision reported more reduction of life quality. CONCLUSION: We found that male subject, temporary habitants, low educational status and no eyewear are risk factors of open globe injury in Shanghai. Occupational injury is the leading cause. CART analysis presents a certain agreement to the actual visual outcome. The injury imposes negative impact on quality of life especially in no vision cases. The education of eye protection may help to avoid the injury.

Bony orbital maldevelopment after enucleation.

One common belief in ophthalmology is that enucleation at an early age will result in bony orbital maldevelopment and facial asymmetry. However, the age range in which enucleation is associated with risk of orbital maldevelopment and the extent of asymmetry remains controversial. In this study, patients who had undergone unilateral enucleation at different ages without orbital implantation were analysed to investigate bony orbital development after enucleation. A total of 87 Chinese adult patients were included. Their bony orbital volume and orbital aditus area were measured using three-dimensional reconstructive models based on patients' computer tomography scans. The ratio of the parameter values of the affected orbit to the unaffected orbit was calculated and described as the orbital symmetry index. The results showed that the bony orbit grew until approximately 18 years old. Enucleation after that age did not affect the orbit, whereas enucleation before that age led to significant orbital maldevelopment. The relative reduction ranged up to 20% in orbital volume and 17% in the orbital aditus area. The extent of orbital maldevelopment was correlated to the age of enucleation. The symmetry index of orbital volume = -0.0003x(2)  + 0.0159x + 0.8112 (x = the age of enucleation). The symmetry index of the orbital aditus area = -0.0002x(2)  + 0.0119x + 0.8504 (x = the age of enucleation). The regression formulae were used to predict the severity of orbital asymmetry after unilateral enucleation, and evaluate the necessity and efficacy of interventions following enucleation.

Ocular complications of human immunodeficiency virus infection in eastern china.

PURPOSE: To investigate ocular complications in patients with HIV/AIDS in eastern China during the time of highly active antiretroviral therapy (HAART). DESIGN: Prospective study. METHODS: This study was carried out from August 1, 2009 to July 31, 2010. Recruited HIV/AIDS patients underwent a series of surveys and ophthalmologic and laboratory examinations (including CD4 level) at enrollment. RESULTS: In this study, all 787 HIV/AIDS patients (1574 eyes) had a history of HAART. Of these patients, 28.72% (95% CI = 0.26-0.32) had a history of systemic disease and 26.30% (95% CI = 0.23-0.29) had ocular complications. Of these ocular complications, cytomegalovirus retinitis (CMVR) had the highest prevalence (10.6%, 83/787) and ocular microangiopathy had the second-highest prevalence (9.4%, 74/787). Among the patients with CMVR, 16.9% (14/83) suffered from immune recovery uveitis (IRU). Furthermore, 3.4% (27/787) of the recruited AIDS patients had neuro-ophthalmologic disorders. The mean logMAR visual acuity of the group with ocular complications was 0.47 ± 0.64, which was significantly different from the asymptomatic group (0.17 ± 0.39, P < .001). The median CD4 T-cell count of the group with ocular complications is 43 cells/µL, which was significantly different from the asymptomatic group (116.5 cells/µL, P < .001). CONCLUSIONS: The study shows a high rate of treatable ocular complications among patients with HIV/AIDS in eastern China. HIV/AIDS treatment programs in China must be prepared to identify ocular complications and refer patients to the correct treatment facilities.

Functional study on a novel missense mutation of the transcription factor FOXL2 causes blepharophimosis-ptosis-epicanthus inversus syndrome (BPES).

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal dominant disease caused by FOXL2 gene mutations. However, only one missense mutation has been found in family with BPES type I. Here, we report a novel missense mutation in the forkhead domain of the FOXL2 gene (c.340A > G, NM_023067) resulted in the replacement of lysine by glutamic acid at amino acid position 114 of the FOXL2 protein (p.K114E, NP_075555) that was identified in a Chinese family with BPES type I, members of which displayed clinical symptoms such as shortened palpebral fissures, drooping eyelids, a vertical skin fold arising from the lower eyelid, and premature ovarian failure (POF) in affected females. Based on the patients' clinical features and computational analysis of this missense mutation in a three-dimensional structural model, we hypothesised that the mutation might disturb the intermolecular contacts between FOXL2 and the StAR gene. The disturbance of this interaction might contribute to the POF observed in BPES type I patients. We performed subcellular localisation and functional studies and as expected, observed significant nuclear aggregation and cytoplasmic mislocalization of the mutant type protein and loss-of-function was confirmed by electrophoretic mobility shift assays, transcriptional activity assays and quantitative real-time polymerase chain reaction. This functional study on a novel missense mutation has important implications for the molecular analysis of this gene.

FOXL2 mutations in Chinese families with Blepharophimosis syndrome (BPES).

Blepharophimosis syndrome (BPES) is a rare, autosomal dominant disease. Two clinical types of BPES have been distinguished. In BPES type I, an eyelid malformation is associated with infertility in affected females as a result of premature ovarian failure. In BPES type II, eyelid anomalies alone are observed. Mutations of FOXL2, which is a gene encoding a forkhead transcription factor, have recently been shown to cause both types of BPES. Here, we report 1 novel duplication mutation of the FOXL2 gene identified in a large Chinese family affected by type II BPES and 1 less recurrent 17-bp duplication in a large Chinese family affected by BPES of an undetermined type. These new cases give additional support to the previously reported genotype-phenotype correlations and our findings have expanded the spectrum of known mutations of the FOXL2 gene.

Quantitative morphometry of the orbit in Chinese adults based on a three-dimensional reconstruction method.

The three-dimensional (3D) reconstruction technique serves as a practical tool in diagnosis, surgical planning, and outcome prediction of plastic and reconstructive surgery. In our study, the morphologic features of the Chinese adult orbit were described by 11 anatomic parameters using a 3D reconstruction technique. Sixty-four Chinese adults were selected randomly from patients who had undergone craniofacial computed tomography (CT) scans to diagnose conditions other than craniofacial or orbital deformations. The morphologic parameters of orbit such as bony orbital volume, orbital foramen area and orbital rim perimeter were measured on 3D models using this technique. Differences between the two orbits and between the two sexes were investigated. The method of measurement showed high reproducibility of results. No difference between the two orbits was found. There were significant differences between men and women in all anatomic parameters other than orbital height. In men and women, respectively, mean bony orbital volume was 26.02 and 23.32 mL, mean orbital foramen area 11.80 and 11.10 cm², mean orbital rim perimeter 12.65 and 12.20 cm, mean orbital height 33.35 and 33.22 mm, mean orbital width 40.02 and 38.00 mm; mean orbital floor length 47.93 and 46.18 mm, mean orbital roof length 52.93 and 50.89 mm, mean medial orbital wall length 46.43 and 44.41 mm, mean lateral orbital wall length 48.38 and 46.91 mm, mean intraorbital distance 27.18 and 25.11 mm, mean extra-orbital distance 98.77 and 93.69 mm. It is concluded that the measurements of these orbital parameters could be obtained from a 3D reconstruction method. The two orbits were symmetric based on orbital volume and other anatomic parameters. Orbital size was significantly smaller in women than in men; orbital height, however, was similar. The findings of the present study allow for quantification of the orbital features of Chinese adults and provide parameters for preoperative planning and prediction of postoperative outcome.