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Most common methods of cellular analysis employ the top-down approach (investigating proteomics or genomics directly), thereby destroying the cell, which does not allow the possibility of using the same cell to correlate genomics with functional assays. Herein we describe an approach for single-cell tools that serve as a bottom-up approach. Our technology allows functional phenotyping to be conducted by observing the cytotoxicity of cells and then probe the underlying biology. We have developed a droplet microfluidic device capable of trapping droplets in the array and releasing the droplet of interest selectively using microvalves. Each droplet in the array encapsulates natural killer cells (NK cells) and tumour cells for real-time monitoring of burst kinetics and spatial coordination during killing by single NK cells. Finally, we use the microvalve actuation to selectively release droplets with the desired functional phenotype such as for fast and serial killing of target tumour cells by NK cells. From this perspective, our device allows for investigating first interactions and real-time monitoring of kinetics and later cell recovery on demand for single-cell omic analysis such as single-cell RNA sequencing (scRNA), which to date, is primarily based on in-depth analyses of the entire transcriptome of a relatively low number of cells.

Mercury is one of the most common heavy metals and a major environmental pollutant that affects ecosystems. Since mercury and its compounds are toxic to humans, even at low concentrations, it is very important to monitor mercury contamination in water and foods. Although conventional mercury detection methods, including inductively coupled plasma mass spectrometry, atomic absorption spectroscopy, and gas chromatography-mass spectrometry, exhibit excellent sensitivity and accuracy, they require operation by an expert in a sophisticated and fully controlled laboratory environment. To overcome these limitations and realize point-of-care testing, many novel methods for direct sample analysis in the field have recently been developed by improving the speed and simplicity of detection. Commonly, these unconventional sensors rely on colorimetric, fluorescence, or electrochemical mechanisms to transduce signals from mercury. In the case of colorimetric and fluorescent sensors, benchtop methods have gradually evolved through technology convergence to give standalone platforms, such as paper-based assays and lab-on-a-chip systems, and portable measurement devices, such as smartphones. Electrochemical sensors that use screen-printed electrodes with carbon or metal nanomaterials or hybrid materials to improve sensitivity and stability also provide promising detection platforms. This review summarizes the current state of sensor platforms for the on-field detection of mercury with a focus on key features and recent developments. Furthermore, trends for next-generation mercury sensors are suggested based on a paradigm shift to the active integration of cutting-edge technologies, such as drones, systems based on artificial intelligence, machine learning, and three-dimensional printing, and high-quality smartphones.

In this study, Polygonum aviculare L. extract, which has superior antioxidative and cellular membrane protective activity, was loaded onto cell penetrating peptide (CPP) conjugated liposomes to enhance transdermal delivery. The physical characteristics of typical liposomes and CPP-conjugated liposomes containing P. aviculare extract were evaluated. The particle sizes of both liposomes were approximately 150 nm. Whereas the zeta potential of typical liposomes was -45 mV, that of CPP-conjugated liposomes was +42 mV. The loading efficiency of P. aviculare extract in both liposomes was calculated to be about 83%. Fluorescent-labeled liposomes were prepared to evaluate cellular uptake and skin permeation efficiency. Using flow cytometry, we found that CPP-conjugated liposomes improved cellular uptake of the fluorescent dye as compared with the typical liposomes. In addition, the skin permeation of CPP-conjugated liposomes was proved higher than that of typical liposomes by confocal laser scanning microscopy studies and Franz diffusion cell experiments. The improved cellular uptake and skin permeation of the CPP-conjugated liposomes were due to the cationic arginine-rich peptide. In vivo studies also determined that the CPP-conjugated liposomes were more effective in depigmentation and anti-wrinkle studies than typical liposomes. These results indicate that the CPP-conjugated liposomes could be effective for transdermal drug delivery of antioxidant and anti-aging therapeutics.

To evaluate dry eye symptoms, tear film function and ocular surface changes in patients with psoriasis.The Dry Eye Questionnaire and ophthalmic examination including the Schirmer test, tear break-up time, corneal fluorescein test, meibomian gland obstruction and conjunctival impression cytology were assessed in patients with chronic plaque psoriasis. Results were compared between 30 patients and 30 healthy controls.The rate of positive responses in the Dry Eye Questionnaire and staining of corneal fluorescein test were significantly higher in the patients (P = 0.030) than in the controls (P = 0.012). The tear break-up time in patients was significantly lower than in the controls (P0.001). However, there were no differences in the Schirmer test and meibomian gland function between the patients and controls. In the impression cytology analysis, more cell alteration and decreased goblet cell density were observed in the patients (P0.001) compared with those obtained from controls (P = 0.003).The dry eye symptom was more common in patients with psoriasis. In addition, the patients showed a higher tear film instability and significant degeneration on the ocular surface when compared with the normal controls.

Purpose: To evaluate the clinical features and surgical outcomes for primary rhegmatogenous retinal detachments (RDs) in patients with pseudophakia after phacoemulsification. Methods: The medical records of patients with pseudophakia after phacoemulsification and intraocular lens implantation who had undergone surgery for primary rhegmatogenous RDs with a minimum duration of follow-up of 12 months were reviewed retrospectively. Results: A total of 104 patients were enrolled in this study and 106 eyes were analyzed. Post-operative retinal attachment was achieved in 87 of the eyes (82.1%) and the final visual acuities (logarithm of the minimum angle of resolution) were improved to 0.65 ± 0.49 from the baseline measurement of 1.51 ± 1.14 (p < 0.001). Re-operations were performed in 24 of the eyes (22.6%) and there were no visible retinal breaks in 30 of the eyes (28.3%). The failure to identify a retinal break during surgery was associated with a lower rate of retinal reattachment, worse final visual acuity, and a higher rate of re-operation (p = 0.002, p = 0.02, and p = 0.002, respectively). The location of the identified retinal break was more common in the superotemporal quadrant than in the other quadrants. Conclusions: The inability to identify a retinal break during surgery was associated with a poor final outcome. Other factors were less important for the functional and anatomic success in patients with pseudophakic RDs.

Purpose: To compare the effect of an intravitreal injection of bevacizumab alone (IVB) or combined with triamcinolone (IVB/IVT) versus triamcinolone (IVT) in patients with diabetic macular edema (DME). Methods: In this randomized three-arm clinical trial, eligible eyes were assigned randomly to one of the three study arms: the IVB group, 2 injections of 1.25 mg of bevacizumab with 6-week intervals; the IVB/IVT group, 1.25 mg of IVB with 2 mg of IVT, and the IVT group, 2 mg of IVT. The clinical course of best-corrected visual acuity and central macular thickness by optical coherence tomography was monitored for up to 12 months after the initial injection. Results: One hundred eleven eyes of 105 patients with DME completed 12 months of follow-up. The IVB/IVT group and the IVT group showed better visual acuity and reduced central macular thickness at 6 weeks and 3 months, compared with the IVB group (p = 0.041, p = 0.02 at 6 weeks; p = 0.045, p = 0.043 at 3 months, respectively). However, no significant difference in visual acuity and central macular thickness was observed between the three groups at 12 months (p = 0.088, p = 0.132, respectively). The frequency of retreatment was lower in the IVB/IVT and IVT groups during the 12-month period (p < 0.001). No significant differences in visual acuity or central macular thickness were observed between the IVB/IVT and IVT groups during the follow-up. Conclusion: IVB/IVT and IVT showed more pronounced effects during the earlier postinjection period. However, levels of visual acuity or central macular thickness at 12 months were comparable in the three study groups. No beneficial effect of the combination injection was observed.

PURPOSE To evaluate the levels of aqueous humor cytokines in the eyes of patients with central serous chorioretinopathy (CSC) before an intravitreal injection of bevacizumab. METHODS In a prospective interventional trial, 20 eyes of 20 patients with symptomatic CSC of at least 3 months duration and 20 eyes of 20 patients with cataract as control were included. Eyes with CSC received a single intravitreal injection of bevacizumab (1.25 mg/0.05 mL). Aqueous humor samples were collected from patients and controls. Multiplex bead assays were used for measurement of cytokines, including vascular endothelial growth factor and platelet-derived growth factor. RESULTS Similar vascular endothelial growth factor levels and significantly decreased platelet-derived growth factor levels were measured in the aqueous humor of eyes from patients with CSC compared with controls (P = 0.172 and P = 0.004, respectively). There was a significant inverse correlation between the vascular endothelial growth factor and platelet-derived growth factor in patients (r = -0.555; P = 0.026). Interleukin 6, interleukin 8, and monocyte chemoattractant protein-1 were detectable, but not significantly different between the patients and controls. CONCLUSION The results of the current study suggest that platelet-derived growth factor contributes to the pathogenesis of CSC; however, the roles of vascular endothelial growth factor in CSC are unclear and warrant further investigation.

Purpose To evaluate spectral-domain optical coherence tomography (OCT) findings based on preferential hyperacuity perimeter (PHP) in patients after idiopathic epiretinal membrane surgery. Methods Forty eyes of 40 patients undergoing surgery for idiopathic epiretinal membrane were included in the study. The best-corrected visual acuity, PHP, Amsler grid test, and OCT were assessed 3 months after surgery. Results Patients were classified based on the defect of hyperacuity by PHP, as follows: hyperacuity defect (n = 18 eyes) and hyperacuity intact (n = 22 eyes). There was no difference in age, gender, duration of symptoms, and visual acuity. The average macular thickness and inner nuclear layer thickness in the hyperacuity defect group was thicker than the hyperacuity intact group (P = 0.041 and P = 0.045, respectively). A disrupted photoreceptor layer was more common in the hyperacuity defect group compared with the hyperacuity intact group (P = 0.001). Conclusion With regard to metamorphopsia, a hyperacuity defect after epiretinal membrane surgery was associated with inner nuclear layer edema and photoreceptor disarrangement.

We report a case involving bilateral posterior ischemic optic neuropathy with Behçet's disease in a 49-year-old woman. The patient presented because of abrupt visual disturbances. An ophthalmic examination, including fluorescein angiography, visual field testing, and visual-evoked potentials, was performed. Her visual acuity was 20/100 bilaterally and posterior ischemic optic neuropathy was suspected. She was treated with systemic methylprednisolone; however, disc pallor developed and the visual acuity remained 20/200 bilaterally. Optic neuropathy can be bilateral, affect both eyes simultaneously, and lead to severe visual loss despite high doses of systemic steroid treatment. Optic neuropathy in Behçet's disease is very rare. This case provides clinical insight into the neuro-ophthalmologic manifestations of Behçet's disease.

To describe the clinical implications of outer foveolar defects on optical coherence tomography (OCT) in eyes treated surgically for macular hole (MH).Retrospective observational case series.Ninety-six eyes of 93 subjects who had undergone surgery for the treatment of idiopathic MH were included. Clinical data, including OCT prior to and at follow-up visits ranging from 3 to 32 months after surgery, were analyzed. Incidence, risk factors, and clinical outcomes of outer foveolar defect were evaluated.Outer foveolar defects were noted on postoperative OCT in 44 of the 96 eyes (45.8%). The mean preoperative diameter of MH in the eyes (333.5 ± 126.3 μm) evidencing outer foveolar defects was significantly smaller than those that did not (504.2 ± 155.6 μm) (P.0001). The mean disappearance time of the defect was estimated to be 182 days after surgery. The mean postoperative visual acuity (logMAR) improved to 0.40 ± 0.26 from 0.58 ± 0.23 after the disappearance of outer foveolar defects.Outer foveolar defects were detected predominantly after surgery for small MHs. The defect decreases gradually in size and eventually disappears completely approximately 6 months after surgery.

Purpose: The purpose of this study was to determine aqueous vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels in patients with central serous chori-oretinopathy (CSC) before a single intravitreal bevacizumab injection. Methods: Twelve eyes with symptomatic CSC were included. Samples from patients with cataracts served as controls. The levels of VEGF and IL-8 concentrations were measured in aqueous humor and plasma by multiplex bead assays. Results: All patients with CSC showed an improvement in visual acuity and resolved neurosensory detachment after intravitreal bevacizumab injection. The aqueous humor levels of VEGF and IL-8 were not significantly increased in patients with CSC compared with the healthy control group (18.2 ± 24.8 vs. 35.3 ± 28.5 pg/mL, P > 0.05; 2.3 ± 0.4 vs. 2.8 ± 0.3 pg/mL, P > 0.05, respectively). The plasma levels of VEGF and IL-8 in patients with CSC were not different from those in the healthy control group. Conclusion: Vascular endothelial growth factor and IL-8 were not increased in the aqueous humor and plasma of patients with CSC. The effect of intravitreal bevacizumab injection as a treatment for CSC must be fully understood, and the true effect of anti-VEGF treatment in patients with CSC remains to be elucidated.

Aims To compare clinical outcomes for patients with central serous chorioretinopathy (CSC) undergoing either focal laser photocoagulation or photodynamic therapy (PDT). Methods 26 eyes of 26 patients with symptomatic CSC underwent focal laser photocoagulation or half-dose PDT, as appropriate. Best-corrected visual acuity, optical coherence tomography and multifocal electroretinography were assessed prospectively prior to treatment and at 1, 3 and 6 months after treatment. Results 12 eyes in the focal laser group and 14 eyes in the PDT group were evaluated. One month after PDT, all eyes, except one, showed complete absorption of subretinal fluid, whereas five eyes in the focal laser group showed residual subretinal fluid (p=0.022). Visual acuity and parameters of multifocal electroretinography improved from baseline at 1, 3 and 6 months after treatment, without any significant differences between the two groups. However, compared with those in the PDT group at 1 month after treatment, P1 latencies in the first annuli of the focal laser group were delayed (40.1±5.5 ms and 34.9±2.5 ms, respectively; p=0.047). Conclusions Compared with focal laser, half-dose PDT may facilitate earlier resolution of macular detachment and earlier recovery of central retinal function. However, at 3 months after treatment and thereafter, no difference in anatomical and functional recovery was noted between the two modalities of treatment.

Purpose: To investigate the clinical features of pegylated-interferon (PEG-IFN)-associated retinopathy in chronic hepatitis patients. Methods: We examined a consecutive case series of 46 patients who were treated with PEG-IFN for chronic hepatitis C or B between October 2007 and September 2008. All patients underwent regular ophthalmologic examinations every 3 weeks during the 6 months after treatment. Results: Ten of the 46 patients (21.73%) developed retinal abnormalities. PEG-IFN-associated retinopathy occurred a mean of 7.25 ± 10.28 weeks after treatment and manifested itself as cotton wool spots and retinal hemorrhages. All patients, except for 1 with a retinal vein occlusion, recovered without cessation of treatment. Conclusions: PEG-IFN-associated retinopathy in chronic hepatitis patients was reversible, and patients recovered without visual complications; however, 1 patient did present with an irreversible visual impairment. This type of retinopathy is usually asymptomatic, and clinicians should closely observe patients for 3 months after treatment.

To evaluate the efficacy of intravitreal bevacizumab injection in the treatment of central serous chorioretinopathy (CSC). In a prospective interventional case series, 40 eyes of 40 patients with idiopathic CSC at least 3 months in duration were included. Patients were treated with once or twice intravitreal bevacizumab injections initially and completed at least 12 months follow-up. Main outcomes measures were the resolution of neurosensory detachment, best-corrected visual acuity, and findings on fluorescein and indocyanine green angiography. Thirty-three of 40 eyes (82.5%) showed complete absence of subretinal fluid at the macula within 3 months following initial intravitreal bevacizumab injection. Eyes exhibiting resolution of subretinal fluid revealed improvement in fluorescein and indocyanine green angiographic findings. The rate of intense hyperfluorescence on indocyanine green angiography was higher in eyes exhibiting resolution of subretinal fluid than eyes exhibiting incomplete absorption of subretinal fluid after intravitreal bevacizumab (72.7 and 28.5%, respectively, p = 0.039). Intravitreal bevacizumab injections generally resulted in anatomic improvement for CSC and may constitute a therapeutic option in CSC. The effect after intravitreal bevacizumab injection for CSC might be related to the hyperfluorescence on indocyanine green angiography.

Purpose: My aim was to evaluate the effect of intravitreal bevacizumab and to determine the concentrations of inflammatory cytokines in patients with macular edema due to branch retinal vein occlusion (BRVO), according to the site of the occlusion. Methods: In a prospective interventional case series, 46 eyes of patients with macular edema due to major (n = 30) or macular BRVO (n = 16) were included. The patients were treated primarily with intravitreal bevacizumab (1.25 mg/0.05 ml) and completed 12 months of follow-up. Reinjections were performed if optical coherence tomography showed persistent or recurrent macular edema. Aqueous humor samples were collected at the time of initial intravitreal injection. Multiplex bead assays were used for the measurement of cytokines. Results: The visual acuity and central macular thickness were improved significantly in patients with major and macular BRVO at 12 months. During 12 months, the eyes with major BRVO received a mean of 3.8 intravitreal injections, whereas those with macular BRVO received a mean of 1.9 intravitreal injections (p = 0.009). Significantly increased concentrations of IL-6, IL-8 and MCP-1 were observed in the aqueous humor of BRVO patients compared with control samples. Higher concentrations of IL-6, VEGF, PDGF-AA and MCP-1 were measured in the aqueous humor of patients with major BRVO compared with macular BRVO at baseline (p < 0.05). Conclusion: Intravitreal bevacizumab was similarly effective for improving macular edema in patients with major and macular BRVO. However, macular BRVO required fewer injections and had lower cytokine levels in the aqueous humor than major BRVO.

Central serous chorioretinopathy (CSC) is characterized by the development of serous neurosensory retinal detachment at the posterior pole [1]. In the majority of patients, CSC is self-limited, and patients usually have a good visual prognosis. However, in some cases of CSC, patients develop progressive visual loss due to persistent serous retinal detachment, cystoid macular degeneration, or retinal pigment epithelium decompensation [2,3]. Clinicians usually elect to observe patients with acute CSC, because these patients generally show self-remission, and traditional treatments like laser photocoagulation or photodynamic therapy may induce complications [4,5]. However, patients with acute CSC often desire more expedient resolution of their disease. The pathophysiology of CSC remains poorly understood. The advent of indocyanine green angiography, however, has demonstrated choroidal involvement in the disease. It is likely that choroidal hyperpermeability is an early event in the development of symptomatic CSC where, under the appropriate circumstances, it may lead to retinal pigment epithelial detachment followed by neurosensory detachment [5]. Vascular endothelial growth factor (VEGF) has been implicated as the major factor responsible for increased vascular permeability [6]. Recently, bevacizumab (Avastin; Genentech Inc., San Francisco, CA, USA), an antibody to VEGF, has been shown to have anti-permeability properties. Intravitreal injection of bevacizumab (IVBI) has been reported to be associated with visual improvement and reduced neurosensory detachment without adverse events in patients with CSC [7]. In this study, we investigated the effect of IVBI in patients with acute CSC.

The principal markers of diabetic eye disease are the breakdown of the blood-retinal barrier, retinal vasculature integrity, and hemodynamic abnormalities [1]. Clinical and experimental findings suggest that, along with retinal change, choroidal vasculopathy might also play a role in the pathogenesis of diabetic retinopathy. We further propose that it might be related to the severity of diabetic retinopathy. Various choroidal abnormalities including obstruction of the choriocapillaris, vascular degeneration, choroidal aneurysms, and choroidal neovascularization have been reported in previous studies on diabetic eyes [2-4]. Choroidal angiopathy, by contrast, has received relatively scant research attention. Until recently, the choroid could be evaluated only by indocyanine green angiography, laser Doppler f lowmetry, or ultrasound [5]. These techniques, though useful for determining vessel abnormalities or changes in the choroidal blood flow, provide no anatomical information on the retinal pigmentary epithelium or the choroidal layers. Spectral-domain optical coherence tomography (OCT) now allows for high-quality higher-resolution cross-sectional macula imaging. Additionally, enhanced-depth imaging (EDI) software enables highly reliable and reproducible measurement of choroidal thickness. The present study, proceeding on the premise that a better clinical understanding of choroidal damage might enable a more accurate assessment of diabetic eye diseases, measured the choroidal thickness in patients with various stages of diabetic retinopathy and compared the results with those of age-matched healthy individuals.

We report a case of posterior scleritis effectively managed with intravitreal bevacizumab. A 71-year-old woman was diagnosed with posterior scleritis. Although she was initially treated with systemic steroids, her clinical presentation deteriorated. She was then treated with a single intravitreal injection of bevacizumab and aqueous humor collection. The aqueous level of vascular endothelial growth factor prior to the intravitreal injection was 880.51 pg/mL, greater than that in the healthy control group (p < 0.001). One month later, the scleritis was completely resolved, and the patient remained stable during six months of follow-up. Intravitreal bevacizumab appears to be an effective adjuvant therapy for patients with posterior scleritis.