In vivo genome editing via CRISPR/Cas9-mediated homology-independent targeted integration for Bietti crystalline corneoretinal dystrophy treatment.

Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessive chorioretinal degenerative disease without approved therapeutic drugs. It is caused by mutations in CYP4V2 gene, and about 80% of BCD patients carry mutations in exon 7 to 11. Here, we apply CRISPR/Cas9 mediated homology-independent targeted integration (HITI)-based gene editing therapy in HEK293T cells, BCD patient derived iPSCs, and humanized Cyp4v3 mouse model (h-Cyp4v3mut/mut) using two rAAV2/8 vectors via sub-retinal administration. We find that sgRNA-guided Cas9 generates double-strand cleavage on intron 6 of the CYP4V2 gene, and the HITI donor inserts the carried sequence, part of intron 6, exon 7-11, and a stop codon into the DNA break, achieving precise integration, effective transcription and translation both in vitro and in vivo. HITI-based editing restores the viability of iPSC-RPE cells from BCD patient, improves the morphology, number and metabolism of RPE and photoreceptors in h-Cyp4v3mut/mut mice. These results suggest that HITI-based editing could be a promising therapeutic strategy for those BCD patients carrying mutations in exon 7 to 11, and one injection will achieve lifelong effectiveness.

Epstein-Barr virus-positive iris diffuse large B-cell lymphoma detected by metagenomic next-generation sequencing.

PURPOSE: Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) is a relatively rare subtype of DLBCL. Herein, we report a case of a patient with EBV-positive iris DLBCL after undergoing penetrating keratoplasty and discuss its possible pathogenesis. METHODS: A 72-year-old male patient presented to our hospital with progressive blurring of vision in the left eye for the past 4 months. Small white nodular lesions were observed on the iris and retinal surface of the left eye, with a white cloud-like opacity in the vitreous cavity. RESULTS: The patient was eventually diagnosed with EBV-positive iris DLBCL after undergoing pathological and metagenomic tests. After injecting methotrexate in the left vitreous cavity and administering systemic and local antiviral treatments, the ocular lesions disappeared. CONCLUSION: EBV infection, drug immunosuppression, and aging-related immune deterioration may play significant roles in the pathogenesis of EBV-positive iris DLBCL. SYNOPSIS: Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) is a new subtype of DLBCL, which rarely occurs. Herein, we report a case of a patient with EBV-positive iris DLBCL after undergoing penetrating keratoplasty and discuss its possible pathogenesis.

The complete mitochondrial genome of the red-footed tortoise (Chelonoidis carbonarius Spix 1824).

In this study, we report the complete mitochondrial genome of the red-footed tortoise (Chelonoidis carbonarius). The compete mitochondrial genome of C. carbonarius is 16,639 bp in length, including 13 protein-coding genes, 22 tRNA genes, and two rRNA genes. The mitogenome was deposited in NCBI GenBank under the accession number OQ789392. Furthermore, we also constructed a phylogenetic tree of Chelonoidis using eight species. These results will aid the conservation of Chelonoidis from the perspective of genetic evolution.

Interpretable Detection of Diabetic Retinopathy, Retinal Vein Occlusion, Age-Related Macular Degeneration, and Other Fundus Conditions.

Diabetic retinopathy (DR), retinal vein occlusion (RVO), and age-related macular degeneration (AMD) pose significant global health challenges, often resulting in vision impairment and blindness. Automatic detection of these conditions is crucial, particularly in underserved rural areas with limited access to ophthalmic services. Despite remarkable advancements in artificial intelligence, especially convolutional neural networks (CNNs), their complexity can make interpretation difficult. In this study, we curated a dataset consisting of 15,089 color fundus photographs (CFPs) obtained from 8110 patients who underwent fundus fluorescein angiography (FFA) examination. The primary objective was to construct integrated models that merge CNNs with an attention mechanism. These models were designed for a hierarchical multilabel classification task, focusing on the detection of DR, RVO, AMD, and other fundus conditions. Furthermore, our approach extended to the detailed classification of DR, RVO, and AMD according to their respective subclasses. We employed a methodology that entails the translation of diagnostic information obtained from FFA results into CFPs. Our investigation focused on evaluating the models' ability to achieve precise diagnoses solely based on CFPs. Remarkably, our models showcased improvements across diverse fundus conditions, with the ConvNeXt-base + attention model standing out for its exceptional performance. The ConvNeXt-base + attention model achieved remarkable metrics, including an area under the receiver operating characteristic curve (AUC) of 0.943, a referable F1 score of 0.870, and a Cohen's kappa of 0.778 for DR detection. For RVO, it attained an AUC of 0.960, a referable F1 score of 0.854, and a Cohen's kappa of 0.819. Furthermore, in AMD detection, the model achieved an AUC of 0.959, an F1 score of 0.727, and a Cohen's kappa of 0.686. Impressively, the model demonstrated proficiency in subclassifying RVO and AMD, showcasing commendable sensitivity and specificity. Moreover, our models enhanced interpretability by visualizing attention weights on fundus images, aiding in the identification of disease findings. These outcomes underscore the substantial impact of our models in advancing the detection of DR, RVO, and AMD, offering the potential for improved patient outcomes and positively influencing the healthcare landscape.

Predictive value of ellipsoid zone-related angle parameters in primary surgery of large macular hole: a case control study.

BACKGROUND: To explore the predictive value of Ellipsoid Zone (EZ) -related angle parameters for the outcome of primary macular hole surgery. METHODS: This was a retrospective study. Patients diagnosed with large macular hole (MH) (minimum diameter > 500 µm) between 2018 and 2021 were enrolled. All patients underwent 25-gauge pars plana vitrectomy, internal limiting membrane (ILM) peeling and air tamponade. Spectral-domain OCT (SD-OCT) and best corrected visual acuity (BCVA) were measured in preoperative and 2-week post-operative. Classic and angle related parameters were measured via ImageJ. Angle regularity (AR) were defined as the standard deviation of the angle parameters in vertical and horizontal direction. RESULTS: Seventy-six eyes were included for analysis; 24 eyes showed an unclosed macular hole at the 2-week postoperative and 52 eyes showed a closed hole. Preoperatively, MLD (P < 0.001), BD (P = 0.009) and diameter of EZ/ELM disruption (P = 0.002 and 0.025) in patients failed to close the hole after primary surgery were significantly larger than those succeeded. EZ-MH (P = 0.018), EZ-NFL (P = 0.006), EZ-GCL (P = 0.004), EZ-INL (P = 0.002), EZ-OPL (P = 0.009) and EZ-ONL (P = 0.011) angles were smaller in patients with unclosed hole. AR of the EZ-NFL (P = 0.009), EZ-GCL (P = 0.009), EZ-OPL (P = 0.023), EZ-ONL (P = 0.048) and Basal-NFL (P = 0.030) angles among the unclosed patients were significantly larger than those of the closed group. EZ-NFL (P = 0.015), EZ-GCL (P = 0.004), EZ-INL (P < 0.001), EZ-OPL (P < 0.001), EZ-ONL (P < 0.001), Basal (P = 0.023) and Basal-NFL (P < 0.001) angles of hole-unclosed patients enlarged significantly after the surgery. CONCLUSIONS: Patients with large macular holes and an increased EZ-related angle and angle AR are more likely to experience unsuccessful outcomes following primary MH surgery. Therefore, EZ-related angles hold potential as valuable parameters for predicting the surgical outcome.

Multimodal Imaging of Choroidal Structural in Torpedo Maculopathy.

Objective: To report a case of torpedo maculopathy with multimodal fundus imaging methods, and apply the choroid vascularity index to quantitatively describe the choroidal structural changes in torpedo maculopathy. Case presentation: An asymptomatic 41-year-old Chinese woman with an incidentally found yellowish-white macular lesion in her left eye was referred to our hospital. She was unaware of any prior medical conditions. The best corrected visual acuity (BCVA) was 20/20 OD and 20/25 OS, respectively. Fundus exam of her left eye revealed a well-circumscribed torpedo-like hypopigmented lesion in the macula region, and the tapered edge directed toward the fovea. Pigment deposition could be seen in the inferotemporal portion of the torpedo lesion. Fluorescein angiography showed the corresponding window defect without leakage and fundus autofluorescence demonstrated low signal throughout the lesion. Enhanced depth imaging optical coherence tomography revealed outer retinal attenuation, subretinal cavitation, subtle inner choroidal excavation and thinning of outer nuclear layer. The diagnosis of torpedo maculopathy was clinically made. Choroidal vascularity index (CVI) and sub-foveal choroidal thickness (SFCT) were applied to display changes of choroidal structure. The results implied that both subfoveal CVI and SFCT of the affected eye seemed relatively lower when compared with the fellow eye. Optical coherence tomography angiography showed reduced density of the choriocapillaris in the temporal area of the lesion and increased capillary density in the nasal area. Functional examinations, including microperimetry, multifocal electroretinogram and static perimetry also revealed reduced retinal sensitivity, decreased stimulated amplitude and suspected scotoma in the lesion area. After 12 months of follow-up, the patient's visual acuity and the clinical appearance of the lesion were unchanged. Conclusion: The torpedo maculopathy may be identified by abnormal appearance with multimodal imaging. Decreased choroidal vascularity in the lesion area measured quantitatively by choroid vascularity index may play a role in pathogenesis of torpedo maculopathy.

Change of termite hindgut metabolome and bacteria after captivity indicates the hindgut microbiota provides nutritional factors to the host.

The gut-dwelling microbiota is an indispensable part of termites. It is influenced by a series of factors, such as diet and captivity. The objectives of this study were to study the metabolic functions of hindgut microbiota and to investigate the influence of captivity on the hindgut microbiota. The dampwood termite Hodotermopsis sjostedti was reared in the laboratory for 6 months. We conducted the metabolome analysis of the fat body from the freshly-collected workers (FBF), the hindgut fluid of the freshly-collected workers (HFF), and the hindgut fluid of laboratory-maintained workers. In addition, the 16S rRNA genes from the hindgut bacteria in the freshly-collected and laboratory-maintained workers were sequenced. According to our results, the concentrations of metabolites associated with amino acid biosynthesis, vitamin biosynthesis, fatty acid biosynthesis, and cofactor biosynthesis were higher in HFF compared with those in FBF, suggesting that the hindgut microbiota provides nutritional factors to the host. However, after captivity, the concentrations of metabolites in the hindgut associated with amino acid biosynthesis, nucleotide sugar metabolism, vitamin biosynthesis, and carbon metabolism decreased, while those associated with the steroid hormone biosynthesis and ovarian steroidogenesis increased. Meanwhile, the 16S amplicon study revealed that the abundance of certain bacteria changed after captivity, such as uncultured Termite Group 1 bacterium, Candidatus Symbiothrix dinenymphae, and unclassified Desulfovibrio. Our findings show that captivity influences the hindgut microbiota and shed light on the metabolic potential of the hindgut microbiota.

Local chronicles reveal the effect of anthropogenic and climatic impacts on local extinctions of Chinese pangolins (Manis pentadactyla) in mainland China.

Anthropogenic and climatic factors affect the survival of animal species. Chinese pangolin is a critically endangered species, and identifying which variables lead to local extinction events is essential for conservation management. Local chronicles in China serve as long-term monitoring data, providing a perspective to disentangle the roles of human impacts and climate changes in local extinctions. Therefore, we established generalized additive models to identify factors leading to local extinction with historical data from 1700-2000 AD in mainland China. Then we decreased the time scale and constructed extinction risk models using MaxEnt in a 30-year transect (1970-2000 AD) to further assess extinction probability of extant Chinese pangolin populations. Lastly, we used principal component analysis to assess variation of related anthropogenic and climatic variables. Our results showed that the extinction probability increased with global warming and human population growth. An extinction risk assessment indicated that the population and distribution range of Chinese pangolins had been persistently shrinking in response to highly intensive human activities (main cause) and climate change. PCA results indicated that variability of climatic variables is greater than anthropogenic variables. Overall, the factors causing local extinctions are intensive human interference and drastic climatic fluctuations which induced by the effect of global warming. Approximately 28.10% of extant Chinese pangolins populations are confronted with a notable extinction risk (0.37 ≤ extinction probability≤0.93), specifically those in Southeast China, including Guangdong, Jiangxi, Zhejiang, Hunan and Fujian Provinces. To rescue this critically endangered species, we suggest strengthening field investigations, identifying the exact distribution range and population density of Chinese pangolins and further optimizing the network of nature reserves to improve conservation coverage on the landscape scale and alleviate human interference. Conservation practices that concentrate on the viability assessment of scattered populations could help to improve restoration strategies of the Chinese pangolin.

Effects of spatially heterogeneous warming on gut microbiota, nutrition and gene flow of a heat-sensitive ungulate population.

Effects of climate warming on trophic cascades are increasingly reported for large herbivores occupying northern latitudes. During the last 40 years, moose (Alces alces) in northeast China have lost nearly half of their historical distribution through their habitat shifting northwards. There are many possible causes of bottom-up and top-down effects of temperature and for moose in northeast China they are poorly understood. Of particular relevance are the effects of extrinsic environmental factors on gene flow, nutritional adaptions, and gut microbiota that occur as moose populations retreat northwards. We combined molecular biology, nutritional ecology and metagenomics to gain deeper mechanistic insights into the effects of temperature on moose populations. In this study, we revealed that the direction and intensity of gene flow is consistent with global warming driving retreats of moose populations. We interpret this as evidence for the northward movement of moose populations, with cooler northern populations receiving more immigrants and warmer southern populations supplying emigrants. Comparison across latitudes showed that warmer late spring temperatures were associated with plant community composition and facilitated related changes in moose protein and carbohydrate intake through altering forage availability, forage quality and diet composition. Furthermore, these nutrient shifts were accompanied by changes in gut microbial composition and functional pathways related to nutrient metabolism. This study provided insights into mechanisms driving effects of spatial heterogeneous warming on genetic, nutritional and physiological adaptions related to key demographic rates and patterns of survival of heat-sensitive ungulates along a latitude gradient. Understanding such changes helps to identify key habitat areas and plant species to ensure accurate assessment of population status and targeted management of moose populations.

Influence of Pleistocene climate fluctuations on the demographic history and distribution of the critically endangered Chinese pangolin (Manis pentadactyla).

BACKGROUND: Pleistocene climate fluctuations have strongly modified species genetic diversity and distributions. The Chinese pangolin (Manis pentadactyla) has been recognized as a critically endangered animal due to heavy poaching and trafficking. However, the effect of Pleistocene climate fluctuations on the genetic diversity and spatial distribution of the Chinese pangolin remains largely unknown. Here, we combined whole genome sequencing data, analysis of complete mitochondrial genomes, and a large amount of occurrence data from field surveys to infer the ancestral demographic history and predict the past spatial dynamics of the Chinese pangolin in Guangdong Province, China. RESULTS: Our results indicated that there were two subpopulations, which showed similar trends of population size change in response to past climatic changes. We estimated a peak effective population size (Ne) during the last interglacial (LIG), followed by a marked decrease (~ 0.5 to fivefold change) until the last glacial maximum (LGM) and a rebound to a small peak population size during the Mid-Holocene (MH). The estimated time of the separation event between two subpopulations was approximately 3,000-2,500 years ago (ka). We estimated that the distribution of suitable areas shrank by 14.4% from the LIG to LGM, followed by an expansion of 31.4% from the LGM to MH and has been stable since then. In addition, we identified an elevational shift and suitable area decreased significantly during the LGM, but that the geographic extent of suitable areas in the western region increased from the LIG to present. The eastern region of Guangdong Province had the highest habitat suitability across all the climate scenarios. CONCLUSIONS: Our results suggested that Pleistocene climate fluctuations played an important role in shaping patterns of genetic diversity and spatial distribution, and that human stressors likely contributed to the recent divergence of two Chinese pangolin subpopulations sampled here. We argue that a key protected area should be established in the eastern region of Guangdong Province. As such, this study provides a more thorough understanding of the impacts of Pleistocene climate fluctuations impacts on a mammalian species in southern China and suggests more robust management and conservation plans for this Critically Endangered species of special interest.

Effect of intravitreal ranibizumab pretreatment on vitrectomy in young patients with proliferative diabetic retinopathy.

BACKGROUND: Younger patients who underwent vitrectomy for proliferative diabetic retinopathy (PDR) display more aggressive nature distinguished from the older patients. Preoperative anti-VEGF therapy has been widely used as an adjunct for PDR surgery. However, the effect of anti-VEGF administration in young diabetics has rarely been evaluated in previous studies. The purpose of this study was to evaluate the effects of ranibizumab pretreatment on vitrectomy surgery in young patients with PDR. METHODS: This was a prospective nonrandomized comparative study. Young patients (<40 years old) undergoing diabetic vitrectomy with or without ranibizumab pretreatment (25 eyes in each group) were analyzed in this study. The use of the drug was determined by the patients' own preference. The two surgical groups were matched according to a complexity score. Intravitreal injection of ranibizumab (IVR) was performed 3-5 days prior to the vitrectomy surgery in the IVR group. Intraoperative records including total surgical time, intraoperative bleeding, the use of endodiathermy, the frequency of relaxing retinotomies, the incidence of iatrogenic retinal breaks, and the use of perfluorocarbon liquid (PFCL) and silicone oil tamponade, and postoperative indices regarding recurrent vitreous hemorrhage (VH), neovascular glaucoma (NVG), recurrent retinal detachment, and visual outcome were evaluated between the two groups. All patients were followed up for one year after surgery. RESULTS: In young PDR patients, the severity of intraoperative bleeding was significantly lower in the IVR group than in the control group (P=0.04). The total surgical time was shorter in the IVR group than in the control group. However, the rate of relaxing retinotomy, the incidence of iatrogenic retinal breaks and the use of PFCL and silicone oil tamponade were not affected by IVR pretreatment but affected by the complexity score of the case. Early postvitrectomy hemorrhage occurred less frequently in the IVR group than in the control group (P<0.001), Early visual recovery was better in the IVR group than in the control group (P=0.03). However, there were no significant differences in the development of late recurrent VH, NVG, recurrent retinal detachment, and final visual outcome. CONCLUSIONS: IVR pretreatment is a safe and effective adjunct to vitrectomy in reducing intraoperative and early postvitrectomy bleeding and should be suggested in young PDR patients. However, IVR does not reduce the incidence of intraoperative and late postoperative complications in these patients. The risk of iatrogenic retinal breaks and silicone oil use are closely correlated with the complexity score of the surgical cases.

Macro-Nutritional Adaptive Strategies of Moose (Alces alces) Related to Population Density.

The distribution area of moose in China has been shrinking back toward the north and northeast because of climate change and human disturbance, and the population number has been declining. Between 2011 and 2015, we studied moose at six sites in the northeast of China during the snowy seasons. We collected fecal samples and plant samples that were used to estimate population densities for moose, as well as their macro-nutrient selection. Out of a total of 257 fecal samples collected at six sites, we identified a total of 120 individual moose (57 females and 63 males). The population density (moose/km2 ± SE) was highest at Hanma with 0.305 ± 0.064 moose/km2 and lowest at Meitian with only 0.028 ± 0.013 moose/km2. Forage availability was different among sites, with the lowest availability at Mohe (58.17 number/20 m2) and highest was Zhanhe (250.44 number/20 m2). Moose at Zhanhe, Hanma, and Nanwenghe had a balanced diet with higher N:C (1:7), while at Meitian, Shuanghe and Mohe the N:C was 1:8. Our results indicate that the southern areas had low forage quality and quantity and this may be the reason for the distribution of the population of moose shrinking northward.

Comparison of the safety and efficacy of triple sequential therapy and transscleral cyclophotocoagulation for neovascular glaucoma in the angle-closure stage.

To compare the efficacy and safety of triple therapy combining intravitreal injection of anti-vascular endothelial growth factor, trabeculectomy, and pan-retinal photocoagulation via binocular indirect ophthalmoscopy, with that of transscleral cyclophotocoagulation (TCP) to treat neovascular glaucoma in the angle-closure stage. Eighteen triple therapy patients and 25 TCP patients between May 2014 and May 2016 were retrospectively analysed. Anterior chamber puncture and anti-VEGF intravitreal injection were performed on the first day of sequential therapy. Trabeculectomy was performed 3-5 d after injection; pan-retinal laser photocoagulation via binocular indirect ophthalmoscopy was initiated 5-7 d later. The IOP of the triple therapy group was lower than that of the TCP group (15.2 ± 2.2 vs. 20.0 ± 8.5 mmHg) and fewer anti-glaucoma drugs were used (0.5 ± 1.0 vs. 0.6 ± 1.0) after treatment. The success rates of the two groups were 89% and 60% respectively (P = 0.032). The visual function of 94% of triple therapy patients was preserved or improved compared to 64% of TCP patients with statistical significance (P = 0.028). No patient in the triple therapy group showed hypotony or eyeball atrophy. Compared to TCP, triple therapy shows higher success rate, fewer complications, and attributes to visual function preservation.

Effects of interspecific interaction-linked habitat factors on moose resource selection and environmental stress.

Resource selection of herbivores is a complex ecological process that operates in relation to biological or non-biological factors, which may affect the feeding and movement, and subsequently their spatial distribution and environmental stress. Here, we estimated moose (Alces alces cameloides) resource selection for habitat variables and the effect of interspecific interactions related to roe deer (Capreolus pygargus bedfordi) on its population distribution and environmental stress in the Khingan Mountain region of northeast China at local and regional scales. Different response patterns of moose resource selection, spatial distribution, and environmental stress to interspecific interaction-linked habitat factors were shown at the two scales. A general ecological chain, response of moose to interspecific interaction-linked habitat factors, was exhibited at the regional scale, and at the local scale, heterogeneous responses, linkages of habitat selection and environmental stress of moose population might be driven by different interspecific interaction patterns. Our study firstly suggested that moose resource selection, food availability, diet quality, population density and environmental stress indicators were impacted by interactions with the distribution of other sympatric herbivore species and showed differences in ecological response chains at various spatial scales. These findings are useful for sympatric herbivore assembly conservation, habitat quality monitoring and management.

A comparison of autologous transplantation of retinal pigment epithelium (RPE) monolayer sheet graft with RPE-Bruch's membrane complex graft in neovascular age-related macular degeneration.

PURPOSE: To compare the outcome after choroidal neovascular (CNV) membrane excision and retinal pigment epithelium (RPE) transplantation and make further evaluation of two types of RPE grafts on the visual function in patients with neovascular age-related macular degeneration (AMD), complicated by massive subretinal haemorrhage. METHODS: We retrospectively reviewed 80 patients who underwent CNV membrane excision with or without RPE transplantation. Two types of RPE grafts were adopted, RPE-Bruch's membrane complex graft (subgroup 1) and RPE monolayer sheet graft (subgroup 2). Outcome measures included pre- and postoperative visual acuity score (VAS), clinical findings, complications and recurrence rates. RESULTS: The mean VAS [Early Treatment of Diabetic Retinopathy Study (ETDRS)] in the CNV membrane excision group was 11.06 ± 18.28 at baseline and 14.41 ± 16.86 at follow-up (p = 0.12) in a mean follow-up period of 24.35 ± 9.31 months. While in subgroup 1, VAS increased from 22.62 ± 23.72 to 35.50 ± 24.46 (p = 0.02) in a mean period of 20.63 ± 6.25 months. The percentage of visual acuity (VA) improvement (at least two or more lines changed) in subgroup 1 is 62.5%, which is significantly higher than that in CNV excision group (23.5%), p = 0.02. In subgroup 2, VAS increased from 16.61 ± 27.98 to 29.16 ± 23.80 (p = 0.02) in a mean period of 21.72 ± 11.09 months. The percentage of VA improvement in subgroup 2 is 58.0%, which is also significantly higher than that in CNV excision group (23.5%), p = 0.02. Postoperative VA elevation was comparable between the two subgroups (p = 0.05). Complications including retinal detachment, proliferative vitreal retinopathy and CNV recurrence occurred in both techniques. Central fixation stability was observed in eight eyes in subgroup 1 and five eyes in subgroup 2. CONCLUSIONS: The autologous RPE transplantation can increase the vision of patients with haemorrhagic AMD. Two types of autologous RPE grafts were both effective and comparable in restoring visual function and central fixation stability.

Dependable and Efficient Clinical Molecular Diagnosis of Chinese RP Patient with Targeted Exon Sequencing.

Retinitis pigmentosa (RP) is the most common inherited retinal disease. It is a clinically and genetically heterogeneous disorder, which is why it is particularly challenging to diagnose. The aim of this study was to establish a targeted next-generation sequencing (NGS) approach for the comprehensive, rapid, and cost-effective clinical molecular diagnosis of RP. A specific hereditary eye disease enrichment panel (HEDEP) based on exome capture technology was used to collect the protein coding regions of 371 targeted hereditary eye disease genes, followed by high-throughput sequencing on the Illumina HiSeq2000 platform. From a cohort of 34 Chinese RP families, 13 families were successfully diagnosed; thus, the method achieves a diagnostic rate of approximately 40%. Of 16 pathogenic mutations identified, 11 were novel. Our study demonstrates that targeted capture sequencing offers a rapid and effective method for the molecular diagnosis of RP, which helps to provide a more accurate clinical diagnosis and paves the way for genetic counseling, family planning, and future gene-targeted treatment.

Effect of supplemental lutein and zeaxanthin on serum, macular pigmentation, and visual performance in patients with early age-related macular degeneration.

PURPOSE: To compare the 2-year effect of multiple doses of lutein/zeaxanthin on serum, macular pigmentation, and visual performance on patients with early age-related macular degeneration (AMD). METHODS: In this randomized, double-blinded, and placebo-controlled trial, 112 early AMD patients randomly received either 10 mg lutein, 20 mg lutein, a combination of lutein (10 mg) and zeaxanthin (10 mg), or placebo daily for 2 years. Serum concentration of lutein/zeaxanthin, macular pigment optical density (MPOD), visual functions including best-spectacle corrected visual acuity (BCVA), contrast sensitivity (CS), flash recovery time (FRT), and vision-related quality of life (VFQ25) was quantified. RESULTS: Serum lutein concentration and MPOD significantly increased in all the active treatment groups. Supplementation with 20 mg lutein was the most effective in increasing MPOD and CS at 3 cycles/degree for the first 48 weeks. However, they both significantly increased to the same peak value following supplementation with either 10 mg or 20 mg lutein during the intervention. No statistical changes of BCVA or FRT were observed during the trial. CONCLUSIONS: Long-term lutein supplementation could increase serum lutein concentration, MPOD, and visual sensitivities of early AMD patients. 10 mg lutein daily might be an advisable long-term dosage for early AMD treatment.

Changes following supplementation with lutein and zeaxanthin in retinal function in eyes with early age-related macular degeneration: a randomised, double-blind, placebo-controlled trial.

AIMS: To investigate functional and macular pigment (MP) changes in patients with early age-related macular degeneration (AMD) after multiple supplementation with lutein and zeaxanthin. METHODS: 112 patients with early AMD were randomly (1:1:1:1) assigned to receive 10 mg lutein, 20 mg lutein, lutein (10 mg)+zeaxanthin (10 mg), or placebo daily for 2 years. MP optical density (MPOD) was recorded at baseline, 48 weeks and 2 years. Retinal sensitivities were measured by multifocal electroretinogram for peak-to-trough amplitude (N1P1) at baseline and at 48 weeks, and in terms of microperimeter-determined mean retinal sensitivity (MRS) at 48 weeks and 2 years. RESULTS: Supplementation with lutein and zeaxanthin augmented MPOD significantly in active treatment groups (all p<0.05). N1P1 response densities showed significant increases in ring 1 and ring 2 after 48 weeks of supplementation, while no significant changes were seen in rings 3-6. Significant increases in MRS were detected after supplementation with either 10 or 20 mg lutein, whereas no such increases were seen in the placebo arm. CONCLUSIONS: Supplementation with lutein and/or zeaxanthin increases MPOD, and supplemental lutein enhances retinal sensitivity, in patients with early AMD. TRIAL REGISTRATION NUMBER: Clinicaltrials.gov NCT10528605.

Anatomical outcome of vitreoretinal surgery using temporary keratoprosthesis and replacement of the trephined corneal button for severe open globe injuries: one-year result.

In this case series of 74 patients with coexisting vitreoretinal injury and severe corneal opacification, after temporary keratoprosthesis (TKP) assisted pars plana vitrectomy (PPV), an allograft corneal transplant was not performed at the same time; instead, the patient's trephined corneal button was sutured back. One year after the surgery, if intraocular pressure of the injured eyes was above 8 mmHg, removing silicone oil was attempted, and penetrating keratoplasty could be performed. Finally, 10 eyes (13.5%) were enucleated due to atrophia bulbi; 46 eyes (62.2%) were silicone-oil sustained; 15 eyes (20.3%) were anatomically restored; and 3 eyes (4.0%) experienced recurrent retinal detachment. These figures only demonstrate a small percentage of the injured eyes in our series, which have PKP indications. It is a practical option to suture back the patient's trephined cornea following a TKP assisted PPV; keratoplasty was reserved for selected cases.

Autologous transplantation of simple retinal pigment epithelium sheet for massive submacular hemorrhage associated with pigment epithelium detachment.

PURPOSE: To evaluate the long-term outcome of autologous simple RPE sheet transplantation in patients with simultaneous massive submacular hemorrhage and pigment epithelium detachment (PED). METHODS: Fourteen patients with 10 occult AMD eyes and 4 PCV eyes underwent a surgical procedure including a 180° peripheral temporal retinotomy, choroidal neovascular membrane (CNVM) excision, and transplantation of an autologous simple RPE sheet developed from the PED region outside the CNVM lesion. Best-corrected visual acuity, multifocal ERG, and microperimetry were recorded to evaluate macular function. Optical coherence tomography, fundus fluorescein angiography, and an RPE autofluorescence test were performed to observe the status of the transplanted simple RPE sheet. RESULTS: Mean follow-up duration was 24 ± 6 months. Mean ETDRS score increased from 14.0 ± 23.4 preoperatively to 31.9 ± 23.8 at 18 months postoperatively (P < 0.05). Of 14 eyes, 5 (36%) presented with central fixation postoperatively, whereas only 3 (21%) eyes presented with central fixation preoperatively. Ten grafts (71%) showed normal RPE color. Postoperative complications included one (7%) retinal detachment due to proliferative vitreoretinopathy and one (7%) delayed recurrent submacular hemorrhage. CONCLUSIONS: Transplantation of an autologous simple RPE sheet for eyes with massive submacular hemorrhage associated with PED was achieved with a simplified surgical procedure, resulting in increased postoperative visual acuity.