PURPOSE:To establish and validate an artificial intelligence (AI)-assisted automatic cataract grading program based on the Lens Opacities Classification System III (LOCS III).SETTING:Eye and Ear, Nose, and Throat Hospital, Fudan University, Shanghai, China.DESIGN:AI training.METHODS:Advanced deep-learning algorithms, including Faster R-CNN and ResNet, were applied to the localization and analysis of the region of interest. An internal dataset from the EENT Hospital of Fudan University and an external dataset from the Pujiang Eye Study were used for AI training, validation, and testing. The datasets were automatically labeled on the AI platform regarding the capture mode and cataract grading based on the LOCS III.RESULTS:The AI program showed reliable capture mode recognition, grading, and referral capability for nuclear and cortical cataract grading. In the internal and external datasets, 99.4% and 100% of automatic nuclear grading, respectively, had an absolute prediction error of ≤1.0, with a satisfactory referral capability (area under the curve [AUC]: 0.983 for the internal dataset; 0.977 for the external dataset); 75.0% (internal dataset) and 93.5% (external dataset) of the automatic cortical grades had an absolute prediction error of ≤1.0, with AUCs of 0.855 and 0.795 for referral, respectively. Good consistency was observed between automatic and manual grading when both nuclear and cortical cataracts were evaluated. However, automatic grading of posterior subcapsular cataracts was impractical.CONCLUSIONS:The AI program proposed in this study showed robust grading and diagnostic performance for both nuclear and cortical cataracts, based on LOCS III.

Purpose:To assess the efficacy, safety, and predictability of presbyopia-correcting intraocular lenses (IOLs) in cataract patients with previous corneal refractive surgery.Methods:A systematic literature search was performed to identify studies evaluating the clinical outcomes of presbyopia-correcting IOLs implantation in cataract surgery after laser refractive surgery. Outcomes were efficacy, safety and predictability parameters.Results:The authors identified 13 studies, involving a total of 128 patients and 445 eyes. Presbyopia-correcting IOLs were effective at improving distance, intermediate and near visual acuity aftercataract surgery. The proportion of post-laser surgery eyes with uncorrected distance visual acuity (UDVA) ≥ 20/25 was 0.82 [95% confidence interval (CI), 0.74-0.90] and the pooled rates of spectacle independence at near, intermediate, and far distances were 0.98 (95% CI, 0.94-1.00), 0.99 (95% CI, 0.95-1.00) and 0.78 (95% CI, 0.65-0.94) respectively. The percentage of participants who suffered from halos and glare was 0.40 (95% CI, 0.25-0.64) and 0.31 (95% CI, 0.16-0.60), respectively. The predictability had a percentage of 0.66 (95% CI, 0.57-0.75) and 0.90 (95% CI, 0.85-0.96) of eyes within ±0.5 diopters (D) and ±1.0 D from the targeted spherical equivalent.Conclusions:Presbyopia-correcting IOLs provide satisfactory results in terms of efficacy, safety and predictability in patients with previous corneal refractive surgery, but have a higher risk of photopic side effects such as halos and glare.

Background: Patients with type 2 diabetes mellitus (T2DM) are prone to ocular surface infections. We therefore characterized the conjunctival microbiome of T2DM patients and the influence of topical levofloxacin to investigate whether a dysbiosis is associated with this phenomenon. Methods: Conjunctival microbiome of 79 T2DM patients and 113 non-diabetic controls was profiled using the 16S rDNA sequencing approach. Furthermore, 21 T2DM and 14 non-diabetic patients who underwent cataract surgeries were followed up perioperatively and the influence of pre- and post-operative levofloxacin on the conjunctival microbiome was further investigated prospectively and compared longitudinally. Results: The α-diversity of the conjunctival microbiota was significantly higher in T2DM patients than in controls ( P < 0.05). Significant differences in both composition and function of the conjunctival microbiome were identified on the ocular surface of T2DM patients as compared to non-diabetic controls. Particularly, phylum Bacteroidetes and Fusobacteria , genus Pseudomonas, Haemophilus , and Empedobacter were enriched, while genus Streptococcus was reduced on the T2DM ocular surface. Microbial genes functioning of bacterial chemotaxis was elevated in the conjunctival microbiome of T2DM patients. Furthermore, compared to the initial status, several genera including Staphylococcus were more abundant in the conjunctival microbiome of T2DM patients after 3-days use of preoperative levofloxacin topically, while no genus was more abundant in the non-diabetic follow-up group. No difference was observed between initial status and 7 days after ceasing all postoperative medications in both diabetic and non-diabetic follow-up groups. Conclusions: The conjunctival microbiome of T2DM patients was more complex and may respond differently to topical antibiotics.

Increasing evidence has shown a correlation between chronic periodontitis (CP) and Alzheimer’s disease (AD). Nevertheless, there is still a lack of direct evidence, and especially key molecules to connect the two diseases. This study aims to investigate potential protein links between CP and AD within the inflammatory aspect. The hippocampus of CP model mice and controls were collected, and changes in protein expression were evaluated using two-dimensional differential in-gel electrophoresis (2D-DIGE) analysis combined with liquid chromatography tandem mass spectrometry. A total of 15 differentially expressed proteins were identified in CP model mice, as compared with the controls. Among them, S100-A9, transthyretin, Cofilin 2, peroxiredoxin 2, and lipocalin-2 were validated by Western blot according to their dual function both in inflammation and AD. Based on 2D-DIGE analysis, CP animal model had higher levels of S100-A9, Cofilin 2, peroxiredoxin 2, and lipocalin-2 compared to controls. The level of Cofilin 2, one of the well-established proteins in the pathology of AD, was strongly correlated with the time course of CP pathology, indicating a specific molecular correlation between CP and AD. Moreover, the in vivo results showed the level of Cofilin 2 increased significantly along with a prominent increase of the phosphorylation of protein phosphatase 2 (PP2A) and tau protein in the cell lysates of Porphyromonas gingivalis (P.g-LPS)-treated SK-N-SH APPwt cells. Cofilin 2 inhibition resulted in a sharp decrease in PP2A dependent of tau phosphorylation. Furthermore, tumor growth factor (TGF)-β1 was one of the most important inflammatory cytokines for the Pg-LPS-induced Cofilin 2 upregulation in SK-N-SH APPwt cells. These results showed inflammation served as the bond between CP and AD, whereas inflammatory related proteins could be the key linkers between the two diseases. Determining the association between CP and AD at the molecular mechanism will not only hold the direct evidence of the association between the two diseases but also provide a new way of preventing and treating AD: the effective prevention and treatment of CP could serve as a useful method to alleviate the development of AD.

目的 了解云南省临沧市小学生近视现状,分析近视的相关影响因素,为当地近视防控提供初步的参考依据.方法 横断面研究.采用随机分层抽样方法,于2019年6月随机抽取云南省临沧市4所小学的一年级至六年级小学生共666名,对调查的小学生进行电脑验光,问卷调查学生的一般情况、每天近距离学习用眼时间、每天电子产品使用时间、每天户外活动时间、近视是否配镜治疗.采用χ2检验比较性别、年龄、年级、民族、每天学习用眼时间、每天电子产品使用时间、每天户外活动时间不同的近视患病率差异,二项logistic回归分析近视的相关影响因素.结果 本次调查小学生平均年龄(9.5±1.6)岁,其中男性343名、女性323名.近视学生人数达277名,近视患病率高达41.6％,而佩戴眼镜者仅占近视学生的18.4％.χ2检验结果显示,女生近视患病率高于男生(χ2=4.617,P=0.032);年龄越大患病率越高(χ2=17.848,P<0.001),年级越高患病率越高(χ2=32.514,P<0.001);每天学习用眼时间越长患病率越高(χ2=6.515,P=0.038),每天电子产品使用时间越久患病率越高(χ2=15.369,P<0.001),每天户外活动时间越久患病率越低(χ2=29.448,P<0.001).不同民族间的近视患病率差异无统计学意义(χ2=6.679,P=0.246).二项logistic回归分析结果显示,小学生近视与否与性别、年龄、年级、每天学习用眼时间、每天电子产品使用时间、每天户外活动时间具有相关性(P均<0.05).结论 云南省临沧市小学生近视患病率高达41.6％,而佩戴眼镜者仅占近视学生的18.4％,是不容忽视的公共健康问题.近视与小学生的性别、年龄和年级相关,同时与小学生的用眼行为密切相关.

51岁女性,因"左眼反复视力下降2年,加重1年"就诊.2年前因左眼反复视力下降,外院诊断为左眼玻璃体积血,予左眼玻璃体切除术,术后症状未见改善.此次就诊左眼裸眼视力0.1,矫正无助,眼压有波动,最高到32.4 mmHg(1 mmHg=0.133 kPa),症状在活动时更明显.裂隙灯检查见角膜色素性KP(+),前房Tyndall(+++),前房细胞(+++),颞上方虹膜基质萎缩伴透照缺损,瞳孔欠圆,直径约4 mm,对光反应迟钝,人工晶状体(IOL)倾斜,玻璃体混浊,眼底糊.5年前曾行双眼白内障摘除联合IOL植入术,双眼高度近视病史.诊断为葡萄膜炎-青光眼-前房积血综合征(UGH),需要与白内障术后的非感染性葡萄膜炎、迟发性感染性眼内炎相鉴别.给予巩膜层间缝线固定IOL置换术,术中房角镜下见前房角色素沉着,送检房水病原菌检测阴性,房水涂片见红细胞.术后1年内随访患者眼压稳定,玻璃体混浊消退,视力提高明显.讨论体会:白内障术后远期出现的反复视力下降伴眼压升高,反复前房炎症反应在考虑UGH时,应首先排除迟发性感染性炎症的可能性.无缝线巩膜固定三片式IOL(如Yamane方法)在高度近视患者中需谨慎使用.

AIMS:The canonical Wnt signaling pathway plays an essential role in blood-brain barrier integrity and intracerebral hemorrhage in preclinical stroke models. Here, we sought to explore the association between canonical Wnt signaling and hemorrhagic transformation (HT) following intravenous thrombolysis (IVT) in acute ischemic stroke (AIS) patients as well as to determine the underlying cellular mechanisms. METHODS:355 consecutive AIS patients receiving IVT were included. Blood samples were collected on admission, and HT was detected at 24 hours after IVT. 117 single-nucleotide polymorphisms (SNPs) of 28 Wnt signaling genes and exon sequences of 4 core cerebrovascular Wnt signaling components (GPR124, RECK, FZD4, and CTNNB1) were determined using a customized sequencing chip. The impact of identified genetic variants was further studied in HEK 293T cells using cellular and biochemical assays. RESULTS:During the study period, 80 patients experienced HT with 27 parenchymal hematoma (PH). Compared to the non-PH patients, WNT7A SNPs (rs2163910, P = .001, OR 2.727; rs1124480, P = .002, OR 2.404) and GPR124 SNPs (rs61738775, P = .012, OR 4.883; rs146016051, P < .001, OR 7.607; rs75336000, P = .044, OR 2.503) were selectively enriched in the PH patients. Interestingly, a missense variant of GPR124 (rs75336000, c.3587G>A) identified in the PH patients resulted in a single amino acid alteration (p.Cys1196Tyr) in the intracellular domain of GPR124. This variant substantially reduced the activity of WNT7B-induced canonical Wnt signaling by decreasing the ability of GPR124 to recruit cytoplasmic DVL1 to the cellular membrane. CONCLUSION:Variants of WNT7A and GPR124 are associated with increased risk of PH in patients with AIS after intravenous thrombolysis, likely through regulating the activity of canonical Wnt signaling.

Cartilage acid protein 1 (CRTAC1) encodes a protein containing the Ca2+binding domain, which can promote apoptosis of human lens epithelial cells (HLECs) induced by ultraviolet B radiation. Exosomes secreted from adipose-derived stem cells (ASC-exo) have been used to treat many diseases, but the effect of ASC-exo on cataracts has not been established. We hypothesized that ASC-exo has a therapeutic effect on cataracts by regulating CRTAC1. We established the UVB-induced injured HLECs model to test the interactions between CRTAC1 and miR-10a-5p, and the effect on the Ca2+ level and reactive oxygen species (ROS) generation in apoptotic HLECs. We found that UVB significantly increased the level of CRTAC1 expression and induced HLEC apoptosis, while ASC-exo inhibited the induction of UVB and exosome inhibitor reduced the inhibition of ASC-exo. The qRT-PCR results showed that miR-10a-5p had a low level of expression in cataract lesions, whereas CRTAC1 was highly expressed. There was a negative correlation between the expression of CRTAC1 and miR-10a-5p. ASC-exo reversed UVB-inhibited miR-10a-5p expression and miR-10a-5p negatively regulated CRTAC1. In vitro data showed that miR-10a-5p reversed UVB-induced ROS, apoptosis, and the Ca2+ level in HLECs. Overexpression of CRTAC1 reversed the induction of ASC-exo in UVB-injured HLECs, and low expression of CRTAC1 reversed the induction of miR-10a-5p inhibitor. By upregulating the level of miR-10a-5p expression and downregulating the level of CRTAC1 expression, exosomes from ASCs attenuated UVB-induced apoptosis, ROS generation, and the Ca2+ level in HLECs. Our research provides novel insight into the treatment methods and associated mechanisms underlying cataracts.

Pyroptosis has been found to be related to diverse ocular diseases, including cataract. Abnormal CRTAC1 expression has been reported to involve in cataract formation by affecting cell apoptosis. Whether CRTAC1 regulates pyroptosis in the formation progress of cataract is completely unknown. Here, we aimed to investigate the regulatory effects of CRTAC1 on pyroptosis and the potential mechanism in the UVB-induced cell damage model. The results showed that the levels of the established pyroptosis markers (NLRP3, active Caspase-1, pro Caspase-1, GSDMD-N, IL-1β and IL-18) were significantly increased in cataract patients. The above pyroptosis markers could be obviously induced by UVB-irradiation in human lens epithelial cells (HLECs), while down-regulation of CRTAC1 significantly reversed the UVB-induced pyroptosis. Up-regulation of CRTAC1 promoted HLECs pyroptosis, while the ROS inhibitor N-acetyl-l-cysteine blocked the effects of CRTAC1 overexpression. In conclusion, our findings further suggested that the prominent role of CRTAC1 in cataract formation.

The apoptosis of human lens epithelial cells (HLECs) is a characteristic change that occurs during the development of cataracts. Ultraviolet B (UVB) is known to induce the generation of reactive oxygen species (ROS) and apoptosis in HLECs, and thus cause cataracts. Previously, we reported the functions of cartilage acidic protein 1 (CRTAC1) in UVB-treated HLECs. However, the underlying mechanism was not known. In this study, we found that CRTAC1 expression and nuclear factor-kappa B (NF-κB) p65 nuclear translocation were elevated in capsule tissues of cataract patients in comparison with normal controls. The NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), alleviated UVB-induced apoptosis in HLECs; while activation of NF-κB suppressed the effects of the ROS inhibitor, N-acetyl-L-cysteine (NAC), on UVB-treated HLECs. The expression and promoter activity of CRTAC1 was inhibited by PDTC and NAC. Moreover, the suppressed effects of CRTAC1 knockdown on UVB-induced ROS generation, cell apoptosis, nuclear translocation of NF-κB p65, and p38 phosphorylation were attenuated by a p38 agonist. In contrast, the p38 inhibitor abolished the promotional effects of CRTAC1 overexpression on HLECs. Taken together, our results for the first time show that NF-κB is a potential transcription factor for CRTAC1. The regulatory network involving NF-κB, CRTAC1, and p38 may therefore play an important role in cataract formation.

Background To investigate the decentration and tilt of plate-haptic multifocal intraocular lenses (MfIOLs) in myopic eyes. Methods Myopic (axial length [AXL] > 24.5 mm) and non-myopic (21.0 mm < AXL ≤ 24.5 mm) cataract eyes were enrolled in this prospective study and randomly assigned to receive implantation of Zeiss AT LISA tri 839MP lenses (Group A) or Tecnis ZMB00 lenses (Group B). In total, 122 eyes of 122 patients were available for analysis. Decentration and tilt of MfIOLs, high-order aberrations (HOAs), and modulation transfer functions (MTFs) were evaluated using the OPD-Scan III aberrometer 3 months postoperatively. Subjective symptoms were assessed with a Quality of Vision questionnaire. Results Near and distance visual acuities, tilt and horizontal decentration did not differ between the two groups, postoperatively. However, myopic eyes of Group B showed greater vertical decentration than those of Group A (− 0.17 ± 0.14 mm vs. -0.03 ± 0.09 mm, respectively), particularly when the MfIOLs were placed horizontally or obliquely. Overall decentration of myopic eyes was greater in Group B than in Group A (0.41 ± 0.15 mm vs. 0.16 ± 0.10 mm, respectively). In Group B, AXL was negatively correlated with vertical decentration and positively correlated with overall decentration. No such correlations were found in Group A. Intraocular total HOAs, coma, trefoil and spherical aberrations were lower in Group A than in Group B for a 6.0 mm pupil among myopic eyes. Generally, Group A had better MTFs and fewer subjective symptoms than Group B among myopic eyes. Conclusions Plate-haptic design of MfIOLs may be a suggested option for myopic cataract eyes due to the less inferior decentration and better visual quality postoperatively.

Increasing evidence indicates Chronic Periodontitis (CP) is a comorbidity of Alzheimer's disease (AD), which is the most common form of age-related dementia, and for the latter, effective diagnostic and treatment strategies are lacking. Although inflammation is present in both diseases, the exact mechanisms and cross-links between CP and AD are poorly understood; and a direct association between the two has not been reported. This study aimed to identify a direct serum proteins link between AD and CP. Two-dimensional differential in-gel electrophoresis was employed to analyze serum samples from 12 CP patients and 12 age-matched controls. Furthermore, to determine the molecular link between CP and AD, neuroblastoma SK-N-SH APPwt cells were treated with 1 mu g/ml of lipopolysaccharide fromPorphyromonas gingivalis(P.g-LPS). Ten differentially expressed proteins were identified in CP patients. Among them, nine proteins were up-regulated, and one protein was down-regulated. Of the 10 differentially expressed proteins, five proteins were reportedly involved in the pathology of AD: Cofilin-2, Cathepsin B, Clusterin, Triosephosphate isomerase, and inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4). Western blotting indicated significantly higher expression of Cofilin-2, Cathepsin B, and Clusterin and lower expression of ITI-H4 in the CP group than in the Control group. The serum concentration of Cathepsin B has a good correlation with MMSE scores. Moreover, the protein level of Cathepsin B (but not that of ADAM10 and BACE1) increased significantly along with a prominent increase in A beta(1-40)and A beta(1-42)in the cell lysates of P.g-LPS-treated SK-N-SH APPwt cells. Cathepsin B inhibition resulted in a sharp decrease in A beta(1-40)and A beta(1-42)in the cell lysates. Furthermore, TNF-alpha was one of the most important inflammatory cytokines for the P.g-LPS-induced Cathepsin B upregulation in SK-N-SH APPwt cells. These results show that CP and AD share an association, while Cathepsin B could be a key link between the two diseases. The discovery of the identical serum proteins provides a potential mechanism underlying the increased risk of AD in CP patients, which could be critical for elucidating the pathophysiology of AD.

PURPOSE To compare the rotational stability of a plate-haptic toric intraocular lens (IOL) versus a C-loop haptic toric IOL in myopic cataract eyes. SETTING EENT Hospital of Fudan University, China. DESIGN Prospective, randomized, controlled study. METHODS Cataract eyes with axial length (AXL)>24.5 mm were randomly assigned to receive implantation of a C-loop haptic toric IOL (AcrySof Toric IOL, Group A) or a plate-haptic toric IOL (AT TORBI 709M, Group B). IOL rotation, residual astigmatism (RAS), visual acuity and high-order aberrations (HOAs) evaluated with OPD-Scan III aberrometer were compared at 3 months postoperatively. In total, 62 eyes of 62 patients were eligible for analysis: 31 in Group A and 31 in Group B. RESULTS The mean rotation of toric IOLs was greater in Group A than in Group B (8.00±3.60° vs 4.42±3.24°, respectively, p<0.001), especially when IOLs were vertically placed. IOL rotation was positively correlated with AXL in Group A while no such correlations were found in Group B. RAS in Group A was greater than that in Group B (-0.76±0.30D vs -0.51±0.29D, respectively, p=0.001). Fewer eyes achieved a RAS of ≤0.50D in Group A than in Group B (38.71% vs 64.52%). Group A had worse postoperative uncorrected visual acuity and higher total HOAs and coma for a 6mm pupil than Group B, while postoperative corrected visual acuity was not different between the two groups. CONCLUSIONS The plate-haptic toric IOL may be a better choice for myopic cataract eyes with corneal astigmatism due to reduced postoperative rotation.

ABSTRACT Purpose To evaluate the necessity for second-eye cataract surgery in bilateral highly myopic patients with good visual acuity in the unoperated fellow eye. Materials and Methods Seventy-five bilateral highly myopic patients who underwent uneventful sequential cataract surgery (the eye with worse visual acuity operated first) were included in this prospective study. The preoperative BCVA of the eye with better visual acuity in these patients was ≤ 0.3 (logMAR, or Snellen ≥ 20/40). Binocular single vision examinations were performed 1 month after the first-eye and second-eye surgery, respectively. The VF-14 questionnaire was completed before first-eye surgery, 1 month after the first-eye and 1 month after the second-eye surgery, respectively. Results The first eye’s postoperative SE was −3.07 ± 1.10D and the second eye’s preoperative SE was −12.91 ± 5.15D. Binocular single vision functional parameters improved significantly after the second-eye cataract surgery compared with that after the first-eye surgery (all P < .001). The binocular single vision function was negatively correlated with the SE difference between the two eyes after the first-eye surgery. No difference was found between the scores of VF-14 questionnaire preoperatively and after the first-eye surgery. VF-14 score only improved significantly after the second-eye surgery compared with that after the first-eye surgery (P < .001). Conclusion In bilateral highly myopic cataract patients, binocular single vision function could improve significantly after the second-eye cataract surgery compared with after the first-eye surgery. Bilateral highly myopic patients may undergo second-eye cataract surgery earlier, even if cataract in that eye is not severe enough to affect the visual acuity.

目的 建立一个稳定、高效检测眼内液中铜绿假单胞菌的体系.方法 采用环介导等温扩增(LAMP)技术,由0.1～0.2 mL眼内平衡盐溶液(BSS)模拟眼内房水或玻璃体,利用铜绿假单胞菌gbca基因,特异性设计5组LAMP引物,并对扩增方法进行关键试剂的优化,检测眼内液中铜绿假单胞菌.结果 该扩增方法经过优化后,最低可以检出100 fg/μL纯基因组DNA,接近荧光定量灵敏度,具有很好的重现性.此外该方法在检测眼内液中混合的菌落时呈现出良好的检测效果,可以检测出2.1×102 CFU/mL,极具应用价值.结论 建立了一个稳定、高效、灵敏的恒温扩增体系,可以应用于眼内液中铜绿假单胞菌的检测.

Purpose: To compare the accuracy of the Barrett Universal II, Haigis, and Olsen formulas in calculating intraocular lens (IOL) power in eyes with extreme myopia. Setting: Eye and Ear, Nose, and Throat Hospital, Fudan University, Shanghai, China. Design: Prospective case series. Methods: Eyes were divided into 3 axial length (AL) groups as follows: 26.0 to 28.0 mm (control), 28.0 to 30.0 mm (extreme myopia 1), and 30.0 mm or more (extreme myopia 2). The mean error (ME) 1 month postoperatively was adjusted to zero by optimizing the lens factor; then, the median absolute errors (MedAEs) were compared between formulas. Factors associated with postoperative refractive errors were analyzed. Results: After optimization, the MEs of the Barrett Universal II, Haigis, and Olsen formulas were 0.04 diopter (D) 0.48 (SD), 0.04 +/- 0.66 D, and 0.04 +/- 0.52 D, respectively, and the MedAEs were 0.37 D, 0.46 D, and 0.39 D, respectively (P = .044; Haigis versus Barrett: P = .038). In the extreme myopia 1 group, all 3 formulas produced small MedAEs (P = .662). In the extreme myopia 2 group, the Haigis formula produced a significantly greater MedAE than the Barrett Universal II formula (P = .007; Haigis versus Olsen: P = .055). The accuracy of the Haigis formula in myopic eyes was affected by the AL and keratometry value, whereas the accuracy of the Barrett Universal II and Olsen formulas was affected by the AL only. Conclusions: In eyes with an AL of 28.0 to 30.0 mm, all 3 formulas were accurate. In eyes with AL of 30.0 mm or more, the Barrett Universal II formula was better than the Haigis formula, possibly because there were fewer influencing factors. (C) 2019 ASCRS and ESCRS

A nuclear cataract is an age-related chronic and priority ophthalmic disease in which a clouding of the lens in the human eye affects vision. Automatic segmentation of nuclear region based on slit-lamp photographs is a basic step for computer-aided diagnosis such as nuclear cataract grading. However, slit-lamp photographs collected from a clinic scenario often have complex background containing the eyelids, sclera and cornea with spectral highlights. The existing efforts using traditional image processing that have unsatisfactory results, and the deep learning method using standard Faster R-CNN tends to obtain a bigger nuclear contour. In this paper, we propose a coarse-to-fine deep learning solution to localize nuclear regions by cascading the Faster R-CNN in a two-stage framework. First, a nuclear ROI (region of interest) predictor is pre-trained to localize a rough position and remove complex backgrounds. Then, a fine nuclear locator is applied to predict a more compact nuclear bounding box. Finally, an ellipse-like nuclear contour is fitted based on its bounding box. Evaluated on a clinical dataset of 884 slit-lamp photographs, the proposed method outperforms the state-of-the-art, improving the overlapping rate (IoU) by 0.33% from 67.98% to 68.31%, and increasing the success rate by 2.55% from 85.71% to 88.26%.

Objective To investigate the effect of a novel Wnt surrogate protein (Wnt-S) on the Wnt/β-catenin sig-naling pathway in human brain microvascular endothelial cells,and may provide a new strategy for the protection of blood-brain barrier (BBB). Methods Wnt-S in pcDNA3. 1( +) plasmid was constructed by molecular cloning technology,and was transfected into HEK293F cells to express and purify Wnt-S protein;Western blot tested Wnt-S with a 5x His-tag;Hu-man brain endothelial cells (hCMEC/D3) marker CD31,VE-cadherin,GLUT1 and MFSD2A were detected by immunofluo-rescence staining;Quantitative real-time PCR was performed to detect the expression of Wnt receptors FZD1-10 and LRP5/6 in HEK293 cells and hCMEC/D3 cells,and the expressions of target gene AXIN2 downstream of the Wnt/β-catenin signa-ling pathway in HEK293 cells and hCMEC/D3 cells after Wnt-S treatment. Results Nucleic acid electrophoresis results showed that Wnt-S plasmid construction was completed;Western blot’s results showed that Wnt-S protein was secreted and detected;FZD1-7 and LRP6 were highly expressed in HEK293 cells, FZD2,4,6 and LRP6 were highly expressed,and FZD1,3 were poorly expressed in hCMEC/D3;Quantitative real-time PCR results showed that AXIN2expression level was significantly up-regulated in HEK293 and hCMEC/D3 cells after addition of Wnt-S protein ( P<0. 001),indicating that Wnt-S protein had biological activity and activated the Wnt/β-catenin signaling pathway. Conclusion Similar to natural Wnt3a protein,Wnt-S protein has a biological function and can activate the canonical Wnt signaling pathway in human cere-brovascular endothelial cells,and thus may protect the BBB.