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The cell type-specific molecular pathology of post-stroke cognitive impairment (PSCI) in the hippocampus has not been thoroughly elucidated. We analyzed 27,069 cells by using single-cell RNA sequencing, and four oligodendrocyte precursor cell (OPC) subtypes were identified, Vcan+ OPCs, which were determined to be the primary cluster among them. Additionally, we examined the features of endothelial cells (ECs) and found that Lcn2+ ECs might play neuroprotective roles via Vwf after stroke. These results may facilitate further studies attempting to identify new avenues of research and novel targets for PSCI treatment.

BackgroundThis study proposed a computational method to detect the cancer areas and calculate the tumor proportion score (TPS) of PD-L1 immunohistochemistry (IHC) expression for lung adenocarcinoma based on deep learning and transfer learning.Patients and methodsPD-L1 22C3 and SP142 IHC slides of lung adenocarcinoma samples on digitized whole-slide images (WSI) database were employed. We build a deep convolutional neural network (DCNN) to automatically segment cancer regions. TPS was calculated based on segmented areas and then compared with the interpretations of pathologists.ResultsWe trained a DCNN model based on 22C3 dataset and fine-tuned it with SP142 dataset. We obtain a robust performance on cancer region detection on both datasets, with a sensitivity of 93.36% (22C3) and 92.80% (SP142) and a specificity of 93.97% (22C3) and 89.25% (SP142). With all the coefficient of determinations larger than 0.9 and Fleiss’ and Cohen’s Kappa larger than 0.8 (between mean or median of pathologists and TPS calculated by our method), we also found out the strong correlation between the TPS estimated by our computational method and estimation from multiple pathologists’ interpretations of 22C3 and SP142 respectively.ConclusionWe provide an AI method to efficiently predict cancer region and calculate TPS in PD-L1 IHC slide of lung adenocarcinoma on two different antibodies. It demonstrates the potential of using deep learning methods to conveniently access PD-L1 IHC status. In the future, we will further validate the AI tool for automated scoring PD-L1 in large volume samples.

We generate arbitrary 1-dimensional spatial profiles in a laser pulse by mapping the temporal electrical waveform sent to an acousto-optic modulator (AOM). The AOM can be therefore used as a spatial light modulator with 45 µm pixel pitch, fast refresh rate, and high damage threshold.

MotivationOncogenes are genes whose malfunctions play critical roles in cancer development, and their discovery is a major aim of cancer mechanisms study. By counting the mutation frequency, oncogenes have been identified with frequent mutations, while it is believed that many more oncogenes could be discovered by differential mutational profile analysis. However, it is common that current methods only utilize mutations in the cancer population, which have an obvious bias in background mutation modelling.MethodsTo predict oncogenes efficiently, we developed a method, DGAT-onco that analyzed the frequency distribution and functional impacts of mutations in both cancer and natural population. Our method can capture the mutational difference of two population, and provide a comprehensive view of genomics basis underlying cancer development. DGAT-onco was constructed by germline mutations from the 1000 Genomes project and somatic mutations of 33 cancer types from the Cancer Genome Atlas (TCGA) dataset. Its reliability was verified on an independent test set including 19 cancers from other sources.ResultsWe demonstrated that our method is more effective than alternative methods in oncogenes discovering. Using this approach achieves higher classification performance in oncogene discovery than 6 alternative methods, and 22.8% significant genes identified by our method were verified as oncogenes by the Cancer Gene Census (CGC).AvailabilityDGAT-onco is available at https://github.com/zhanghaoyang0/DGAT-onco.Contactyangyd25@mail.sysu.edu.cn or zhaohy8@mail.sysu.edu.cn

Sumoylation is one of the key regulatory mechanisms in eukaryotes. Our previous studies reveal that sumoylation plays indispensable roles during lens differentiation (Yan et al. 2010. Proc Natl Acad Sci USA. 107:21034–21039; Gong et al. 2014. Proc Natl Acad Sci USA. 111:5574–5579). Whether sumoylation is implicated in cataractogenesis, a disease largely derived from aging, remains elusive. In the present study, we have examined the changing patterns of the sumoylation ligases and de‐sumoylation enzymes (SENPs) and their substrates including Pax6 and other proteins in cataractous lenses of different age groups from 50 to 90 years old. It is found that compared with normal lenses, sumoylation ligases 1 and 3, de‐sumoylation enzymes SENP3/7/8, and p46 Pax6 are clearly increased. In contrast, Ubc9 is significantly decreased. Among different cataract patients from 50s to 70s, male patients express more sumoylation enzymes and p46 Pax6. Ubc9 and SENP6 display age‐dependent increase. The p46 Pax6 displays age‐dependent decrease in normal lens, remains relatively stable in senile cataracts but becomes di‐sumoylated in complicated cataracts. In contrast, sumoylation of p32 Pax6 is observed in senile cataracts and increases its stability. Treatment of rat lenses with oxidative stress increases Pax6 expression without sumoylation but promotes apoptosis. Thus, our results show that the changing patterns in Ubc9, SENP6, and Pax6 levels can act as molecular markers for senile cataract and the di‐sumoylated p46 Pax6 for complicated cataract. Together, our results reveal the presence of molecular signature for both senile and complicated cataracts. Moreover, our study indicates that sumoylation is implicated in control of aging and cataractogenesis., The molecular signatures for both senile cataract and complicated cataract. Upregulation of sumoylation ligases Ubc9 and PIAS1, and de‐sumoylation enzyme SENP6, and SUMO1 conjugation of p32 Pax6 are molecular markers for senile cataract. Di‐sumoylation of p46 Pax6 is the molecular marker for complicated cataract.

We generate arbitrary 1-dimensional spatial profiles in a laser pulse by mapping the temporal electrical waveform sent to an acousto-optic modulator (AOM). The AOM can be therefore used as a spatial light modulator with 50 q,m pixel pitch, fast refresh rate, and high damage threshold.

We generate arbitrary 1-dimensional spatial modes of light using an acousto-optic modulator to control the optical wavefront of a laser pulse, realizing a spatial light modulator with 50 µm pixel pitch and fast refresh rate.

Background: The COVID-19 pandemic crisis is posing challenges for clinical practice worldwide. We describe the characteristics and effectiveness of a virtual clinical service run by a tertiary ophthalmic center in China with an aim to provide practical guidance for such services during and beyond the pandemic. Methods: We conducted a longitudinal observational study. To provide the diagnosis and treatment of eye diseases under stringent epidemic control measures, the Zhongshan Ophthalmic Center (ZOC) established a virtual clinical service (virtual 2020) by harnessing several digital technologies (5G telecommunication networks, big data analytics, artificial intelligence (AI) and blockchain technology). Multiple interrelated services were offered online, including AI prescreening, virtual live consultation, and online pharmacy. We extracted data from the online clinical service records, along with on-site face-to-face (F2F) clinical records obtained from February 1 to March 13 in 2020 (F2F 2020) as well as a similar period in 2019 (F2F 2019) for analysis. We analyzed the clinical service pattern by day and time; indication of visit and demographics of age, gender, and geographical origin between three groups: virtual 2020, F2F 2020, and F2F 2019. Findings: A total of 38,038 visits to the ZOC were conducted online and F2F from February 1 to March 13, 2020, including 10,641 visits from an AI chatbot, 9,850 virtual live consultations by 127 doctors, and 17,547 on-site F2F consultations in the clinics. With the gradual opening of on-site outpatient and emergency services, online service numbers steadily increased. The AI chatbot services after office hours (8 pm to 8 am) accounted for 26.9% of the total visits, which was significantly higher than the proportions of virtual live consultations (10.9%) and on-site F2F consultations (3.5%) in 2020. Among the indications for virtual live consultation, specific disease consultation was the most common across all ages (64.7–70.7%); younger patients tended to have visual symptoms (18–34 years old, 60.4%), whereas seniors were more likely to require prescription renewal (>55 years old, 63.5%). Virtual live consultation in 2020 was more common in younger (median age 32 years) and female patients (53.3%) than in the F2F 2020 or F2F 2019 groups. Ocular surface diseases were the most frequent diagnosis in the virtual live consultations, which differed from the top diagnosis in F2F 2020 (retinopathy) and F2F 2019 (refraction) groups (P

Background: Computed tomography (CT) characteristics associated with critical outcomes of patients with coronavirus disease 2019 (COVID-19) have been reported. However, CT risk factors for mortality are poorly understood. We aimed to investigate the automatically quantified CT imaging predictors for COVID-19 mortality. Methods: In this retrospective study, laboratory-confirmed COVID-19 patients at Wuhan Central Hospital between December 9, 2019, and March 19, 2020, were included. A novel prognostic biomarker, V-HU score, depicting the volume of total pneumonia infection and the average Hounsfield unit (HU) value of consolidation areas was quantified from CT by an artificial intelligence (AI) system. Cox proportional hazards models were used to investigate risk factors for mortality. Findings: This study included 238 patients (126 survivors and 112 non-survivors). The V-HU marker was an independent predictor (hazard ratio [HR] 2·78, 95% CI 1·50-5·17; p=0·0012) after adjusting for several COVID-19 prognostic indicators significant in univariable analysis. The prognostic performance of the model containing clinical and outpatient laboratory factors was improved by integrating the V-HU marker (c-index: 0·695 versus 0·728; p =65 years; HR 3·56, 95% CI 1·64-7·71; p=0·0006) and younger patients (age

Under photon-limited detection which is limited by the low-light illumination and short detection time, off-the-shelf classification methods based on clear imaging of the object cannot achieve considerable classification accuracy. To solve this problem, we propose a non-imaging classification method based on single-pixel imaging system. With low-intensity pulsed illumination and time-correlated single-photon counting detection, binarized feature sequence of the objects that need to be classified can be obtained. Combining with a simple machine learning algorithm trained with simulated data based on Poissonian photon detection algorithm, the objects could be classified with considerable accuracy. Proof-of-principle experiments use the MNIST handwriting digit database, showing that up to $$90\%$$ classification accuracy could be achieved with fewer than 1 detected photon per pixel.

The differences in transcription start sites (TSS) and transcription end sites (TES) among gene isoforms can affect the stability, localization, and translation efficiency of mRNA. Isoforms also allow a single gene different functions across various tissues and cells However, methods for efficient genome-wide identification and quantification of RNA isoforms in single cells are still lacking. Here, we introduce single cell Cap And Tail sequencing (scCAT-seq). In conjunction with a novel machine learning algorithm developed for TSS/TES characterization, scCAT-seq can demarcate transcript boundaries of RNA transcripts, providing an unprecedented way to identify and quantify single-cell full-length RNA isoforms based on short-read sequencing. Compared with existing long-read sequencing methods, scCAT-seq has higher efficiency with lower cost. Using scCAT-seq, we identified hundreds of previously uncharacterized full-length transcripts and thousands of alternative transcripts for known genes, quantitatively revealed cell-type specific isoforms with alternative TSSs/TESs in dorsal root ganglion (DRG) neurons, mature oocytes and ageing oocytes, and generated the first atlas of the non-human primate cornea. The approach described here can be widely adapted to other short-read or long-read methods to improve accuracy and efficiency in assessing RNA isoform dynamics among single cells.

Single-pixel imaging has emerged over recent years as a novel imaging technique, which has significant application prospects. In this paper, we propose and experimentally demonstrate a scheme that can achieve single-pixel non-imaging object recognition by acquiring the Fourier spectrum. In an experiment, a four-step phase-shifting sinusoid illumination light is used to irradiate the object image, the value of the light intensity is measured with a single-pixel detection unit, and the Fourier coefficients of the object image are obtained by a differential measurement. The Fourier coefficients are first cast into binary numbers to obtain the hash value. We propose a new method of perceptual hashing algorithm, which is combined with a discrete Fourier transform to calculate the hash value. The hash distance is obtained by calculating the difference of the hash value between the object image and the contrast images. By setting an appropriate threshold, the object image can be quickly and accurately recognized. The proposed scheme realizes single-pixel non-imaging perceptual hashing object recognition by using fewer measurements. Our result might open a new path for realizing object recognition with non-imaging.

The aim of the present study was to investigate the clinical characteristics and the underlying genetic causes of Best vitelliform macular dystrophy (BVMD) in a sporadic case in a Chinese patient. A 10-year-old boy was diagnosed with BVMD; complete ophthalmic examinations were performed, including best-corrected visual acuity, intraocular pressure, slit-lamp examination, fundus photograph, optical coherence tomography and fundus fluorescein angiography imaging. Genomic DNA was extracted from leukocytes of the peripheral blood collected from this patient and his family members. DNA samples from 200 unrelated subjects from the Chinese population were used as controls. A total of 11 exons of the bestrophin 1 (BEST1) gene were amplified by polymerase chain reaction and directly sequenced. The results revealed that the patient presented with yellowish lesions in the macular area. Heterozygous mutations c.292G>A (p.Glu98Lys) in exon 4 and c.1608C>T (p.Thr536Thr) in exon 10 of the BEST1 gene were identified in this sporadic case; however, this was not identified in any of his unaffected family members or in the normal controls. The c.292G>A (p.Glu98Lys) mutation has not been previously reported, whereas the c.1608C>T (p.Thr536Thr) mutation is a previously characterized single nucleotide polymorphism (SNP). In conclusion, BEST1 gene mutations and polymorphisms have been reported in diverse ethnic groups, and the present study identified a novel BEST1 gene mutation and an SNP that occurred simultaneously in a Chinese patient with BVMD.

The current study was performed with aim to investigate the fibroblast growth factor receptor 2 (FGFR2) gene in two Chinese families with two different forms of syndromic craniosynostosis, and to characterize their associated clinical features. Two families underwent complete ophthalmic examinations, and two patients from each family were diagnosed with craniosynostosis. Genomic DNA was extracted from leukocytes of peripheral blood collected from these two families and from 200 unrelated subjects within the same population as controls. Exons 8 and 10 of the FGFR2 gene were amplified by polymerase chain reaction and directly sequenced. Ophthalmic examinations of the two patients revealed shallow orbits and ocular proptosis, accompanied by midface hypoplasia and craniosynostosis. Case 1 had retinal detachment, abnormal limbs and hands, while case 2 exhibited normal hands and feet upon clinical examination. A heterozygous FGFR2 missense mutation c.833G>T (C278F) in exon 8 was identified in these two patients, but not in unaffected family members or the normal controls. Although FGFR2 gene mutations and polymorphisms have been studied in various ethnic groups, we report a mutation of FGFR2 in two different Chinese patients with two different types of syndromic craniosynostosis.

Fibrosis is a chronic process involving development and progression of multiple diseases in various organs and is responsible for almost half of all known deaths. Epithelial–mesenchymal transition (EMT) is the vital process in organ fibrosis. Lens is an elegant biological tool to investigate the fibrosis process because of its unique biological properties. Using gain- and loss-of-function assays, and different lens fibrosis models, here we demonstrated that microRNA (miR)-26a and miR-26b, members of the miR-26 family have key roles in EMT and fibrosis. They can significantly inhibit proliferation, migration, EMT of lens epithelial cells and lens fibrosis in vitro and in vivo. Interestingly, we revealed that the mechanisms of anti-EMT effects of miR-26a and -26b are via directly targeting Jagged-1 and suppressing Jagged-1/Notch signaling. Furthermore, we provided in vitro and in vivo evidence that Jagged-1/Notch signaling is activated in TGFβ2-stimulated EMT, and blockade of Notch signaling can reverse lens epithelial cells (LECs) EMT and lens fibrosis. Given the general involvement of EMT in most fibrotic diseases, cancer metastasis and recurrence, miR-26 family and Notch pathway may have therapeutic uses in treating fibrotic diseases and cancers.

Single-cell transcriptome and single-cell methylome analysis have successfully revealed the heterogeneity in transcriptome and DNA methylome between single cells, and have become powerful tools to understand the dynamics of transcriptome and DNA methylome during the complicated biological processes, such as differentiation and carcinogenesis.Inspired by the success of using these single-cell -omics methods to understand the regulation of a particular "-ome," more interests have been put on elucidating the regulatory relationship among multiple-omics at single-cell resolution. The simultaneous profiling of multiple-omics from the same single cell would provide us the ultimate power to understand the relationship among different "-omes," but this idea is not materialized for decades due to difficulties to assay extremely tiny amount of DNA or RNA in a single cell.To address this technical challenge, we have recently developed a novel method named scMT-seq that can simultaneously profile both DNA methylome and RNA transcriptome from the same cell. This method enabled us to measure, from a single cell, the DNA methylation status of the most informative 0.5-1 million CpG sites and mRNA level of 10,000 genes, of which 3200 genes can be further analyzed with both promoter DNA methylation and RNA transcription. Using the scMT-seq data, we have successfully shown the regulatory relationship between DNA methylation and transcriptional level in a single dorsal root ganglion neuron (Hu et al., Genome Biol 17:88, 2016). We believe the scMT-seq would be a powerful technique to uncover the regulatory mechanism between transcription and DNA methylation, and would be of wide interest beyond the epigenetics community.

Single-cell transcriptome and single-cell methylome analysis have successfully revealed the heterogeneity in transcriptome and DNA methylome between single cells, and have become powerful tools to understand the dynamics of transcriptome and DNA methylome during the complicated biological processes, such as differentiation and carcinogenesis.Inspired by the success of using these single-cell -omics methods to understand the regulation of a particular "-ome," more interests have been put on elucidating the regulatory relationship among multiple-omics at single-cell resolution. The simultaneous profiling of multiple-omics from the same single cell would provide us the ultimate power to understand the relationship among different "-omes," but this idea is not materialized for decades due to difficulties to assay extremely tiny amount of DNA or RNA in a single cell.To address this technical challenge, we have recently developed a novel method named scMT-seq that can simultaneously profile both DNA methylome and RNA transcriptome from the same cell. This method enabled us to measure, from a single cell, the DNA methylation status of the most informative 0.5-1 million CpG sites and mRNA level of 10,000 genes, of which 3200 genes can be further analyzed with both promoter DNA methylation and RNA transcription. Using the scMT-seq data, we have successfully shown the regulatory relationship between DNA methylation and transcriptional level in a single dorsal root ganglion neuron (Hu et al., Genome Biol 17:88, 2016). We believe the scMT-seq would be a powerful technique to uncover the regulatory mechanism between transcription and DNA methylation, and would be of wide interest beyond the epigenetics community.

Purpose This study aims to reveal retinal abnormities in a spontaneous diabetic nonhuman primate model and explore the mechanism of featured injuries. Methods Twenty-eight cynomolgus monkeys were identified to suffer from spontaneous type 2 diabetes from a colony of more than eight-hundred aged monkeys, and twenty-six age-matched ones were chosen as controls. Their blood biochemistry profiles were determined and retinal changes were examined by multimodal imaging, hematoxylin and eosin staining, and immunofluorescence. Retinal pigment epithelium (RPE) cells were further investigated by RNA sequencing and computational analyses. Results These diabetic monkeys were characterized by early retinal vascular and neural damage and dyslipidemia. The typical acellular capillaries and pericyte ghost were found in the diabetic retina, which also exhibited reduced retinal nerve fiber layer thickness compared to controls (all P < 0.05). Of note, distinct sub-RPE drusenoid lesions were extensively observed in these diabetic monkeys (46.43% vs. 7.69%), and complements including C3 and C5b-9 were deposited in these lesions. RNA-seq analysis revealed complement activation, AGE/RAGE activation and inflammatory response in diabetic RPE cells. Consistently, the plasma C3 and C4 were particularly increased in the diabetic monkeys with drusenoid lesions (P = 0.028 and 0.029). Conclusions The spontaneous type 2 diabetic monkeys featured with early-stage retinopathy including not only typical vascular and neural damage but also a distinct sub-RPE deposition. The complement activation of RPE cells in response to hyperglycemia might contribute to the deposition, revealing an unrecognized role of RPE cells in the early-stage pathological process of diabetic retinopathy.

The Tianshan Shengli Tunnel is currently the longest high-altitude highway tunnel. In order to ensure the safe operation of the tunnel, the SES software is used to investigate the wind speed laws in the tunnel, and the tunnel is divided into 16 disaster prevention sections based on the shaft and critical wind speed. The principles of disaster prevention and rescue of Tianshan Shengli Highway Tunnel are given, and the emergency treatment process of tunnel disaster prevention and rescue is given. The disaster prevention and rescue plan of Tianshan Shengli Tunnel from fire occurrence, personnel and vehicle evacuation, and fire treatment are discussed in detail.

Taking advantages of aqueous electrolytes such as high safety, ease of fabrication, and high ion conductivity, a series of aqueous batteries have been reported, including lithium-ion batteries, sodium-ion batteries, zinc-ion batteries, aluminum-ion batteries, and novel batteries such as ammonium batteries. They realize energy storage via the repeated (de)intercalation of charge carriers or conversion, which sets strict criteria for choosing proper electrode hosts. Molybdenum-based materials are very competitive candidates for aqueous battery assembly because of their specific layered/tunnel structure and low cost, but their development in this area remains at the infant stage. This review sums up the latest advances on the use of molybdenum-based materials as electrode materials for aqueous batteries. The main strategies for improving their electrochemical properties are summarized, including the introduction of oxygen/sulfur vacancy, interlayer spacing tuning, substrate coating, and electrolyte formulation. The working principles of these methods are compared and discussed in detail. In addition, the main opportunities, achievements, and challenges in this field are briefly commented.

•Li1·2(Mn0·53Co0.27)O2 was successfully synthesized by microwave sintering method.•The batteries show superior rate capacity and cyclic performance.•Crystal growth during microwave and conventional sintering are compared.•Rapid sintering is studied by in-situ TEM heating to give direct evidences.

Crouzon syndrome, a dominantly inherited disorder and the most common type of craniosynostosis syndrome, is caused by mutations in the fibroblast growth factor receptor 2 (FGFR 2) gene, and characterized by craniosynostosis, shallow orbits, ocular proptosis, midface hypoplasia and a curved, beak-like nose. The purpose of the present study was to investigate the fibroblast growth factor receptor 2 (FGFR 2) gene in two Chinese families with Crouzon syndrome and to characterize the associated clinical features. Two families underwent complete ophthalmic examination, and three patients in two families were diagnosed with Crouzon syndrome. Genomic DNA was extracted from leukocytes of peripheral blood samples, which were collected from the family members and 200 unrelated control subjects from the same population. Exons 8 and 10 of the FGFR 2 gene were amplified using polymerase chain reaction analysis and were directly sequenced. Ophthalmic examinations, including best-corrected visual acuity, slit-lamp examination, fundus examination and Computerized Tomography scans, and physical examinations were performed to exclude systemic diseases. These patients were affected with shallow orbits and ocular proptosis, accompanied by midface hypoplasia, craniosynostosis, strabismus or papilloedema, with clinically normal hands and feet. A heterozygous FGFR 2 missense mutation, c.811-812insGAG (p.273insGlu) in exon 8 was identified in the affected individual, but not in the unaffected family members or the normal control individuals in family 1. In family 2, another heterozygous FGFR 2 missense mutation, c.842A>G (P.Tyr281Cys or Y281C), in exon 8 was identified in the affected boy and his mother, but not in the unaffected family members or the normal control individuals. Although FGFR 2 gene mutations and polymorphisms have been reported in various ethnic groups, particularly in the area of osteology, the present study reported for the first time, to the best of our knowledge, the identification of two novel FGFR 2 gene mutations in Chinese patients with Crouzon syndrome.

Objective— Angiogenesis is tightly controlled by growth factors and cytokines in pathophysiological settings. Interleukin 37 (IL-37) is a newly identified cytokine of the IL-1 family, some members of which are important in inflammation and angiogenesis. However, the function of IL-37 in angiogenesis remains unknown. We aimed to explore the regulatory role of IL-37 in pathological and physiological angiogenesis. Approach and Results— We found that IL-37 was expressed and secreted in endothelial cells and upregulated under hypoxic conditions. IL-37 enhanced endothelial cell proliferation, capillary formation, migration, and vessel sprouting from aortic rings with potency comparable with that of vascular endothelial growth factor. IL-37 activates survival signals including extracellular signal-regulated kinase 1/2 and AKT in endothelial cells. IL-37 promoted vessel growth in implanted Matrigel plug in vivo in a dose-dependent manner with potency comparable with that of basic fibroblast growth factor. In the mouse model of retinal vascular development, neonatal mice administrated with IL-37 displayed increased neovascularization. We demonstrated further that IL-37 promoted pathological angiogenesis in the mouse model of oxygen-induced retinopathy. Conclusions— Our findings suggest that IL-37 is a novel and potent proangiogenic cytokine with essential role in pathophy siological settings.

We isolated peripheral blood mononuclear cells from a patient diagnosed with early-onset non-syndromic retinitis pigmentosa caused by compound heterozygous mutations in CRB1 (NM_001257965): c.1369C>T (p.R457X) and c.2027C>T (p.T676M). These cells were transfected with four inducible plasmids encoding human OCT4, SOX2, KLF4 or C-MYC together. Transfected cells were induced to form pluripotent stem cells (iPSCs) expressing the pluripotent stem cell markers SOX2, OCT4, KLF4, SSEA4 and NANOG and presenting a normal karyotype. These cells could be differentiated into three germ layers as well as retinal organoids, and thus provide a valuable cellular model for the study of development of major retina diseases.

Photon-limited imaging technique is desired in tasks of capturing and reconstructing images by detecting a small number of photons. However, it is still a challenge to achieve high photon-efficiency. Here, we propose a novel photon-limited imaging technique that explores the consistency of photon detection probability in a single pulse and light intensity distribution in a single-pixel correlated imaging system. We demonstrated theoretically and experimentally that our technique can reconstruct a high-quality 3D image by using only one pulse each frame, thereby achieving a high photon efficiency of 0.01 detected photons per pixel. Long-distance field experiments for 100 km cooperative target and 3 km practical target are conducted to verify its feasibility. Compared with the conventional single-pixel imaging, which requires hundreds or thousands of pulses per frame, our technique saves two orders of magnitude in the consumption of total light power and acquisition time.

Photon-limited imaging has significant application under extreme environments, in which the photon efficiency is an important parameter. In this paper, we investigate the photon efficiency of computational ghost imaging with single-photon detection. By exploiting the Poisson statistical characteristics of single-photon counting, the relationships between the photon efficiency and the signal-to-noise ratio of the retrieved image are theoretically obtained, and both are closely related to the distribution characteristics of target and spatial modulation in the ghost imaging framework. To verify the availability of the built theoretical model for the photon efficiency, a specific computational ghost imaging with a binary object and the first-photon imaging algorithm is rigorously demonstrated in theory and in experiment. Our investigation paves the way for optimization of photon-limited ghost imaging.

Background CC-Cruiser is an artificial intelligence (AI) platform developed for diagnosing childhood cataracts and providing risk stratification and treatment recommendations. The high accuracy of CC-Cruiser was previously validated using specific datasets. The objective of this study was to compare the diagnostic efficacy and treatment decision-making capacity between CC-Cruiser and ophthalmologists in real-world clinical settings. Methods This multicentre randomized controlled trial was performed in five ophthalmic clinics in different areas across China. Pediatric patients (aged ≤ 14 years) without a definitive diagnosis of cataracts or history of previous eye surgery were randomized (1:1) to receive a diagnosis and treatment recommendation from either CC-Cruiser or senior consultants (with over 5 years of clinical experience in pediatric ophthalmology). The experts who provided a gold standard diagnosis, and the investigators who performed slit-lamp photography and data analysis were blinded to the group assignments. The primary outcome was the diagnostic performance for childhood cataracts with reference to cataract experts' standards. The secondary outcomes included the evaluation of disease severity and treatment determination, the time required for the diagnosis, and patient satisfaction, which was determined by the mean rating. This trial is registered with ClinicalTrials.gov (NCT03240848). Findings Between August 9, 2017 and May 25, 2018, 350 participants (700 eyes) were randomly assigned for diagnosis by CC-Cruiser (350 eyes) or senior consultants (350 eyes). The accuracies of cataract diagnosis and treatment determination were 87.4% and 70.8%, respectively, for CC-Cruiser, which were significantly lower than 99.1% and 96.7%, respectively, for senior consultants (p

Neurodegenerative diseases are a set of disorders characterized by progressive neuronal death and are associated with microglia-mediated neuroinflammation. Recently, neuroinflammation is proposed as a promising therapeutic target for many neurodegenerative diseases. Alpha-1 antitrypsin (AAT) is recognized as a novel immunomodulatory agent in autoimmune diseases and transplantation, however, its impact on neuroinflammation and neurodegeneration remains unknown. This study aims to explore the effects of AAT on microglia-mediated neuroinflammation and retinal degeneration in rd1 mouse model. We found reduced expression of AAT in rd1 retina, and AAT supplement exhibited certain protective effect on retinal degeneration, presenting with increased amount of photoreceptor nuclei, and amplified wave amplitudes in electroretinogram analysis. Of note, AAT shifted microglia phenotype from pro-inflammatory M1 (CD16/CD32+, iNOS+) to anti-inflammatory M2 (CD206+, Arg1+) both in vivo and in vitro, underscoring the concept of immunomodulation on microglia polarization by AAT during neurodegeneration. Furthermore, AAT suppressed the activation of STAT1, promoted the expression of IRF4 while inhibited IRF8 expression, indicating the involvement of these signaling pathways in AAT immunomodulation. Collectively, our data provided evidence for a novel protective role of AAT through immunomodulation on microglia polarization. Attenuating neuroinflammation by AAT may be beneficial to retard neurodegeneration in rd1 mice.

Accumulating evidence suggests the hippocampus being involved in, and modified with, chronic neuropathic pain. However, it is still not clear whether hippocampal activity has direct control over neuropathic behaviors. Here we show that activation of the dorsal, but not ventral, hippocampus, by glutamate microinjection or by chemogenetically increasing excitability (PSAM/PSEM), completely or partially reversed neuropathic behaviors: tactile allodynia and thermal hyperalgesia in the models of spared nerve injury and lumbar spinal nerve ligation. Using a new methodology (chemo-fMRI), where we combine awake resting state brain imaging with viral vector mediated chemogenetic activation (PSAM/PSEM), we could demonstrate that increased excitability of dorsal hippocampus neurons altered resting state functional connectivity within circuitry specifically related to the extent of diminution of neuropathic behavior (tactile allodynia). The identified circuitry most reliably (survived a validation procedure) identified dorsal hippocampal connections to the somatosensory cortex and the thalamus. Moreover, anterograde tracing indicated non-overlapping projections from dorsal and ventral hippocampus. Thus, the present study exhibits a novel causal role for the dorsal hippocampus, and mediating circuitry, controlling neuropathic pain-related behaviors. Altogether, these results imply downregulation of dorsal hippocampus circuitry in chronic neuropathic pain; the activation of which reverses pain behaviors either through disruption of accumulated memories and/or by enhancing extinction circuitry.

SUMMARYCancer cell survival upon cytotoxic drug exposure leads to changes in cell identity, dictated by the epigenome. Several metabolites serve as substrates or co-factors to chromatin-modifying enzymes, suggesting that metabolic changes can underlie change in cell fate. Here, we show that progression of triple-negative breast cancer (TNBC) to taxane-resistance is characterized by altered methionine metabolism and S-adenosylmethionine (SAM) availability, giving rise to DNA hypomethylation in regions enriched for transposable elements (TE). Compensatory redistribution of H3K27me3 forming Large Organized Chromatin domains of lysine (K) modification (LOCK) prevents expression of TE in taxane-resistant cells. Pharmacological inhibition of EZH2, the H3K27me3 methyltransferase, alleviates TE repression, leading to the accumulation of dsRNA and activation of the interferon viral mimicry-response, specifically inhibiting the growth of taxane-resistant TNBC. Together, our work delineates a role for metabolic adaptations in redefining the epigenome of taxane-resistant TNBC cells and underlies an epigenetic vulnerability toward pharmacological inhibition of EZH2.

Table S1. Primers used for real-time PCR. (DOCX 17Â kb)

Transforming growth factor (TGF)-β2-induced epithelial-mesenchymal transition (EMT) in human retinal pigment epithelium (RPE) cells has an important role in the pathophysiology of intraocular fibrotic disorders, which may cause vision impairment and blindness. Autophagy, an intracellular homeostatic pathway, contributes to the physiological and pathological processes of RPE. Furthermore, autophagy has previously been reported to function in the EMT process in numerous tissue and cell types. However, the association between autophagy and the EMT process in RPE cells has not yet been fully determined. The present study demonstrated that TGF‑β2‑treated human RPE cells (ARPE‑19 cell line) exhibited a significantly increased autophagic flux compared with control cells, as determined by western blot analysis of the protein levels of microtubule‑associated protein 1 light chain 3‑II and p62 (also termed sequestosome 1). Furthermore, it was demonstrated that autophagy activation enhanced the TGF‑β2‑induced EMT process in ARPE‑19 cells, and inhibition of autophagy by chloroquine administration attenuated TGF‑β2‑induced EMT, which was determined by analyzing the expression of mesenchymal and epithelial markers by reverse transcription‑quantitative polymerase chain reaction and/or western blotting. A transwell migration and invasion assays was also performed that demonstrated that autophagy activation by rapamycin enhanced TGF‑β2‑stimulated RPE cell migration and invasion, and inhibition of autophagy reduced TGF‑β2‑stimulated RPE cell migration and invasion. These results also demonstrated that autophagy activation enhanced the TGF‑β2‑induced EMT process in ARPE‑19 cells, and inhibition of autophagy attenuated TGF‑β2‑induced EMT. Overall, the results of the present study demonstrated that TGF‑β2‑induced EMT may be regulated by autophagy, thus indicating that autophagy may serve as a potential therapeutic target for the attenuation of EMT in intraocular fibrotic disorders.

Conventional imaging at low light levels requires hundreds of detected photons per pixel to suppress the Poisson noise for accurate reflectivity inference. We propose a high-efficiency photon-limited imaging technique, called fast first-photon ghost imaging, which recovers the image by conditional averaging of the reference patterns selected by the first-photon detection signal. Our technique merges the physics of low-flux measurements with the framework of computational ghost imaging. Experimental results demonstrate that it can reconstruct an image from less than 0.1 detected photon per pixel, which is three orders of magnitude less than conventional imaging techniques. A signal-to-noise ratio model of the system is established for noise analysis. With less data manipulation and shorter time requirements, our technique has potential applications in many fields, ranging from biological microscopy to remote sensing.

The purpose of the current study was to investigate the 11 bestrophin-1 (BEST1) exons in patients with best vitelliform macular dystrophy (BVMD), and to characterize the associated clinical features. Complete ophthalmic examinations were conducted on two families, and two family members were diagnosed with BVMD. Genomic DNA was extracted from the leukocytes of peripheral blood collected from the patients and their family members, in addition to 100 unrelated control subjects recruited from the same population. The polymerase chain reaction was used to amplify a total of 11 exons of the BEST1 gene, which were directly sequenced. Ophthalmic examinations, including best-corrected visual acuity, slit-lamp examination, fundus examination, fundus photography and fluorescein angiography imaging, as well as anterior segment analysis with Pentacam and optical coherence tomography, were conducted. The patients exhibited yellowish lesions in the macular area. A heterozygous mutation c.910_912delGAT (p.304del Asp) in exon 7 was identified in Case 1. A heterozygous BEST1 missense mutation c.685T>G (p.Trp229Gly) in exon 5 was identified in Case 2, but not in any of the unaffected family members or normal controls. Although BEST1 gene mutations and polymorphisms have previously been reported in various ethnic groups, the current study identified, for the first time to the best of our knowledge, two novel BEST1 gene mutations in patients with BVMD.

Fibrosis is a chronic process involving development and progression of multiple diseases in various organs and is responsible for almost half of all known deaths. Epithelial-mesenchymal transition (EMT) is the vital process in organ fibrosis. Lens is an elegant biological tool to investigate the fibrosis process because of its unique biological properties. Using gain- and loss-of-function assays, and different lens fibrosis models, here we demonstrated that microRNA (miR)-26a and miR-26b, members of the miR-26 family have key roles in EMT and fibrosis. They can significantly inhibit proliferation, migration, EMT of lens epithelial cells and lens fibrosis in vitro and in vivo. Interestingly, we revealed that the mechanisms of anti-EMT effects of miR-26a and -26b are via directly targeting Jagged-1 and suppressing Jagged-1/Notch signaling. Furthermore, we provided in vitro and in vivo evidence that Jagged-1/Notch signaling is activated in TGFβ2-stimulated EMT, and blockade of Notch signaling can reverse lens epithelial cells (LECs) EMT and lens fibrosis. Given the general involvement of EMT in most fibrotic diseases, cancer metastasis and recurrence, miR-26 family and Notch pathway may have therapeutic uses in treating fibrotic diseases and cancers.

IL-37 is a novel pro-angiogenic cytokine that potently promotes endothelial cell activation and pathological angiogenesis in our previous study, but the mechanisms behind the pro-angiogenic effect of IL-37 are less well understood. Extending our observations, we found that TGF-β interacts with IL-37, and potently enhances the binding affinity of IL-37 to the ALK1 receptor complex, thus allowing IL-37 to signal through ALK1 to activate pro-angiogenic responses. We further show that TGF-β and ALK1 are required in IL-37 induced pro-angiogenic response in ECs and in the mouse model of Matrigel plug and oxygen-induced retinopathy. The result suggests that IL-37 induces pro-angiogenic responses through TGF-β, which may act as the bridging molecule that mediates IL-37 binding to the TGF-β receptor complex.

Background/Aims: Platelet-derived growth factors (PDGFs) have emerged as pivotal in pathological angiogenesis, which is a hallmark of various tumors and retinal diseases. Here we evaluated the anti-angiogenic effect of imatinib, an inhibitor of PDGF receptors α and β (PDGFR-α and -β), in retinal neovascularization using an oxygen-induced retinopathy (OIR) model. Methods: The OIR model was established and given imatinib or vehicle treatments daily from P12 to P16. At the peak of angiogenesis at P17, the neovascularization area was quantified on retinal whole-mounts with isolectin B4 staining. Immunofluorescence staining and western blots were used to determine the effect of imatinib on different vascular cells and the pathway molecules involved. Results: Imatinib effectively suppressed pathological angiogenesis in OIR mice and reduced the number of all three types of vascular cells, including endothelial cells, pericytes, and smooth muscle cells. Moreover, the expression and activation of PDGFR-α and -β were inhibited by imatinib. The imatinib-treated OIR mice presented with reduced expression of other potent pro-angiogenic factors such as VEGF and FGF2. No obvious retinal or systemic side effects were observed in the imatinib treatment group. Conclusions: Imatinib appears to be safe and effective in suppressing retinal neovascularization. Targeting PDGFs/PDGFRs may also be important for anti-angiogenic treatment and offer a viable alternative treatment for retinal angiogenic diseases.

Background and objective Lens regeneration is an optimal strategy for cataract patients to regain visual acuity with accommodation. We recently designed a novel, minimally invasive capsulorhexis surgical method for cataract removal that achieved functional lens regeneration in human infants. However, small anterior capsulorhexis requires advanced surgical expertise. To examine whether the quality of the regenerated lens can be maintained with enlarged anterior capsulorhexis, we investigated the shape and transparency of the regenerated lenses with different anterior capsulorhexis diameters (ACDs). Methods Thirty-six 4-week-old New Zealand albino rabbits were randomly divided into three groups which underwent lens extraction with different ACDs (Group A: 2.0±0.5 mm, Group B: 4.0±0.5 mm, Group C: 6.0±0.5 mm). The anterior capsule opening area (ACOA) was quantified, and the morphology, weight, and histological characteristics of the regenerated lenses were examined. Results Lens regeneration was observed in all three groups. In Group A, the regenerated lenses were relatively complete and transparent. In Groups B and C, the regenerated lenses were doughnut-shaped and opaque. The speed of lens regeneration in Group A was significantly faster than that in Groups B and C. The ACOA in Group A healed quickly and completely approximately 2 weeks after surgery. However, in Groups B and C, ACOA did not heal completely until 12 weeks after surgery. Histological examination showed that in Group A, most of the lens epithelial cells differentiated into well-organized lens fibers. However, in Groups B and C, the regenerated lens fibers were disorganized. Conclusion Capsulorhexis size is a critical determinant of integrity and transparency in lens regeneration.

We propose a photon-limited imaging technique, first-photon ghost imaging, which reconstructs image by counting the pulses before the first photon arrives. It can achieve visibility enhancement with

We propose and experimentally demonstrate a non-imaging perceptual hashing algorithm based on ghost imaging system can realize non-imaging recognition of image. This technique could find applications in image recognition.

We propose and experimentally demonstrate object saliency detection by means of Fourier spectrum acquisition scheme can realize single pixel non-imaging object saliency detection. This technique can be applied to saliency detection and recognition of image.

Interleukin-1β (IL-1β) is a potent proinflammatory cytokine that plays a critical role in initiating immunoinflammatory responses. In this study, we generated recombinant mouse IL-1β and anti-mouse IL-1β polyclonal antibodies to examine the effect of IL-1β on experimental autoimmune uveoretinitis (EAU), a mouse model for T cell-mediated eye autoimmune disease. Administration of mouse IL-1β by i.p. in the priming phase, but not in the effector phase, of immune response of EAU enhanced disease scores and its related immune responses including DTH, Ag-specific T cell proliferation and the production of IL-17 and IFN-γ. Furthermore, administration of anti-IL-1β antibody in the priming phase reduced EAU scores. These results suggest that IL-1β is an important mediator in the pathogenesis of autoimmune diseases such as uveitis.

Blood vessel degeneration is critically involved in nearly all types of degenerative diseases. Therefore strategies to enhance blood vessel protection and survival are highly needed. In this study, using different animal models and cultured cells, we show that PDGF-CC is a potent vascular protective and survival factor. PDGF-CC deficiency by genetic deletion exacerbated blood vessel regression/degeneration in various animal models. Importantly, treatment with PDGF-CC protein not only increased the survival of retinal blood vessels in a model of oxygen-induced blood vessel regression but also markedly rescued retinal and blood vessel degeneration in a disease model of retinitis pigmentosa. Mechanistically, we revealed that heme oxygenase-1 (HMOX1) activity is critically required for the vascular protective/survival effect of PDGF-CC, because blockade of HMOX1 completely abolished the protective effect of PDGF-CC in vitro and in vivo. We further found that both PDGF receptors, PDGFR-β and PDGFR-α, are required for the vasoprotective effect of PDGF-CC. Thus our data show that PDGF-CC plays a pivotal role in maintaining blood vessel survival and may be of therapeutic value in treating various types of degenerative diseases.

The proliferation and epithelial–mesenchymal transition (EMT) of retinal pigment epithelium (RPE) cells are the major pathological changes in development of proliferative vitreoretinopathy (PVR), which leads to severe visual impairment. Histone deacetylases (HDACs)-mediated epigenetic mechanisms play important roles in controlling various physiological and pathological events. However, whether HDACs are involved in the regulation of proliferation and EMT in PRE cells remains unidentified. In this study, we evaluated the expression profile of HDAC family (18 genes) and found that some of class I and class II HDACs were up-regulated in transforming growth factor-β2 (TGF-β2)/TGF-β1-stimulated RPE cells. Tricostatin A (TSA), a class I and II HDAC inhibitor, suppressed the proliferation of RPE cells by G1 phase cell cycle arrest through inhibition of cyclin/CDK/p-Rb and induction of p21 and p27. In the meantime, TSA strongly prevented TGF-β2–induced morphological changes and the up-regulation of α-SMA, collagen type I, collagen type IV, fibronectin, Snail and Slug. We also demonstrated that TSA affected not only the canonical Smad signalling pathway but also the non-canonical TGF-β/Akt, MAPK and ERK1/2 pathways. Finally, we found that the underlying mechanism of TSA affects EMT in RPE cells also through down-regulating the Jagged/Notch signalling pathway. Therefore, this study may provide a new insight into the pathogenesis of PVR, and suggests that epigenetic treatment with HDAC inhibitors may have therapeutic value in the prevention and treatment of PVR.

Highly-dispersed Fe3O4@ZrO2 yolk–shell structures with a ZrO2 shell of homogeneous shell thickness was successfully prepared via a polymer surfactant (hydroxypropyl cellulose) assisted sol–gel method. By using HPC as a surfactant, highly dispersed particles with Zirconia shells of about 25–30 nm thickness were obtained. The yolk–shell Fe3O4@ZrO2 structure was characterized by several techniques, including transmission electron microscopy (TEM), scanning electron microscopy, (SEM) and X-ray diffraction (XRD), which indicated that the particles had a ZrO2 shell 500 nm in diameter and 25–30 nm in thickness, and a Fe3O4 core of 300 nm in diameter. An in situ TEM heating experiment from 20 °C to 1000 °C demonstrated that the obtained yolk–shell Fe3O4@ZrO2 structure was stable, without any distinguishable structural damage below 700 °C. This material has great potential as a high temperature stable microwave absorber. Even at temperatures of 500 °C, this material still preserved over 90% of its reflection loss (RL) value compared to its room temperature properties. These findings may shed light on the development of novel microwave absorbers for high temperature operation.

The treatment of pediatric cataracts includes surgical and conservative interventions. The ultimate goal of both treatment strategies is to improve visual functions of pediatric patients to the maximum extent and thereby enhance their quality of life. This chapter briefly reviews the history of pediatric cataract surgeries, indicating that the operative methods are constantly evolving with the development of surgical techniques and devices. However, surgery is not the only treatment option for pediatric cataracts. Conservative treatment may be considered for pediatric patients when the lens opacity is mild or the visual axis is not affected. For premature infants or children complicated with developmental disorders or systemic diseases, conservative treatment should also be considered. This chapter also introduces lens regeneration as a treatment of infantile cataracts—a new approach in pediatric cataract management.

The unique characteristics of pediatric eyes, such as thin and elastic sclera, shallow anterior chamber, extremely elastic lens capsule, and high posterior chamber pressure not only enhance the operative difficulty of lens surgery but also increase the risk for intraoperative complications. Prevention and proper management of intraoperative complications would help to ensure more favorable surgical outcomes. Owing to the development and application of modern phacoemulsification techniques and devices in pediatric cataract surgery, the safety of pediatric lens surgery has been greatly improved; however, even a single improper procedure at any stage of the operation may lead to the occurrence of complications. Based on the anatomical and physiological characteristics of pediatric eyes, this chapter will analyze in detail the cause, prevention, and management of potential complications at each step of cataract surgery, including incision construction, capsulorhexis, aspiration of lens materials, intraocular lens implantation, posterior capsulotomy, and vitrectomy.