Pulsed Ultrasound-Mediated Enhancement on Transscleral and Transconjunctival Fluorescein Sodium Delivery to Rabbit Eye In Vivo.

Purpose: To investigate the efficacy and safety of pulsed ultrasound (PUS) in enhancing fluorescein sodium (NaF) transport to the rabbit eye through the transscleral and transconjunctival routes in vivo. Methods: PUS and NaF were applied onto the supratemporal sclera/conjunctiva of healthy rabbit eyes. PUS (1 MHz, 2.37 W/cm2, 30% duty cycle, 5-min application time) was performed 3 times with a 5-min interval. In the same process, NaF was administered to the eye without PUS in the control. NaF concentrations in the vitreous and retina-choroid were determined by fluorescence measurement. The safety of PUS application was assessed based on temperature and intraocular pressure measurements, clinical observations, electroretinography, histology, and Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling assay. Results: In comparison to the control, higher NaF concentrations were found in the retina-choroid following transscleral (2.45-fold) and transconjunctival (2.97-fold) PUS applications (P < 0.05). NaF concentrations in the vitreous were 3.15 and 5.86 times greater in transscleral and transconjunctival PUS applications, respectively, compared with those obtained without PUS application (P < 0.05), and NaF level in the vitreous after transconjunctival PUS application was 2.61 times that of transscleral PUS application (P < 0.05). Ocular findings were transient and mild conjunctival injection, with no other structural and functional changes in PUS-treated eyes. Conclusions: PUS treatment can improve transscleral and transconjunctival delivery of NaF efficiently and safely. Transscleral and transconjunctival PUS applications offer potential clinical benefit in increasing drug penetration to the posterior segments of the eye for the noninvasive treatment of ocular diseases.

Optic Nerve Regeneration in Diabetic Retinopathy: Potentials and Challenges Ahead.

Diabetic retinopathy (DR), the most common microvascular compilation of diabetes, is the leading cause of vision loss and blindness worldwide. Recent studies indicate that retinal neuron impairment occurs before any noticeable vascular changes in DR, and retinal ganglion cell (RGC) degeneration is one of the earliest signs. Axons of RGCs have little capacity to regenerate after injury, clinically leading the visual functional defects to become irreversible. In the past two decades, tremendous progress has been achieved to enable RGC axon regeneration in animal models of optic nerve injury, which holds promise for neural repair and visual restoration in DR. This review summarizes these advances and discusses the potential and challenges for developing optic nerve regeneration strategies treating DR.

Directly induced human retinal ganglion cells mimic fetal RGCs and are neuroprotective after transplantation in vivo.

Retinal ganglion cell (RGC) replacement therapy could restore vision in glaucoma and other optic neuropathies. We developed a rapid protocol for directly induced RGC (iRGC) differentiation from human stem cells, leveraging overexpression of NGN2. Neuronal morphology and neurite growth were observed within 1 week of induction; characteristic RGC-specific gene expression confirmed identity. Calcium imaging demonstrated γ-aminobutyric acid (GABA)-induced excitation characteristic of immature RGCs. Single-cell RNA sequencing showed more similarities between iRGCs and early-stage fetal human RGCs than retinal organoid-derived RGCs. Intravitreally transplanted iRGCs survived and migrated into host retinas independent of prior optic nerve trauma, but iRGCs protected host RGCs from neurodegeneration. These data demonstrate rapid iRGC generation in vitro into an immature cell with high similarity to human fetal RGCs and capacity for retinal integration after transplantation and neuroprotective function after optic nerve injury. The simplicity of this system may benefit translational studies on human RGCs.

The effects of walking training onset on motor evoked potentials after acute spinal cord injury.

AIM: The purposes of this study were to explore the optimal time for starting walking training in the first month after spinal cord injury (SCI) in rats and to discuss the relationship between changes in motor function and transcranial electrical motor evoked potentials (tceMEPs). METHODS: Four groups of rats with SCI (BSWTT-3, 7, 14, 12) performed body-weight-supported treadmill training (BWSTT) for three weeks beginning at 3, 7, 14, and 21 days after SCI, respectively. The Basso, Beattie, and Bresnahan (BBB) score and tceMEPs were assessed weekly. Weekly repeated measures and multiple comparisons between groups were performed to identify differences in motor function and tceMEPs. Correlation analysis was performed to clarify the relationship between BBB scores and tceMEPs over time. RESULTS: Although there was no significant difference between the BWSTT-14 group and the other three BWSTT groups at the end of the experiment in terms of BBB scores and the latency of tceMEPs, the BWSTT-14 group obtained the best trends in improvement of BBB scores and the latencies and amplitudes of tceMEPs over time. The BBB scores of rats with SCI were strongly negatively correlated with tceMEPs latency. The BBB scores of rats with SCI, except for the rats in the BWSTT-3 group, were strongly positively correlated with the tceMEPs amplitude. CONCLUSIONS: The preliminary conclusion was that based on a rat model, 14 days after SCI was the optimal time for starting BWSTT. tceMEPs were an objective indicator of spinal cord nerve function, which was strongly correlated with motor function recovery. However, one limitation of this study was that the rats in the Sham group did not undergo BWSTT, which made the training itself a confounding factor for the results.

A modified fabrication procedure of retinal explant and optimized formulation of culture medium in a three-dimensional retinal culture system.

BACKGROUND: Three-dimensional culture system of retinal explant is commonly used to study retinal ganglion cell (RGC) axon regeneration in vitro. The retinal explants fabricated by traditional procedure in culture system, however, are usually too small (merely 0.5 × 0.5 mm) to be easily detected or treated by current experimental techniques. Also, the constituents of culture medium have not been fully elucidated. NEW METHOD: A fabrication procedure was developed to enlarge the retinal explants and explore the reasonable concentration of fetal bovine serum (FBS) for evaluating axonal regeneration. RESULTS: There were no significant differences in the density or length of regenerative neurites in the retinal explants fabricated by traditional and modified procedures. Increased FBS concentrations promoted neurite regeneration, decreased RGCs apoptosis, and activated tyrosine kinase B (TrkB) receptors, all reaching a plateau at 1 % FBS. COMPARISON WITH EXISTING METHODS: Compared with traditional procedure, the modified fabrication procedure facilitates application of experimental techniques to retinal explants, increases the efficiency of obtaining observation area of regenerating neurites, and reduces the wastage of retinal tissues. The recommended FBS concentration determined in this study is shown to be more suitable for studying neuronal regeneration. CONCLUSION: The retinal explants made by the modified fabrication procedure are successfully applied to the three-dimensional culture system, and presented several advantages over the traditional one. Furthermore, a preliminary experiment must be performed to determine the suitable concentration of FBS in each study to ensure accuracy and stability of the results obtained from the three-dimension retinal culture system.

Design and Evaluation of a Novel Laminectomy Auxiliary Device Based on Rats.

Most types of spinal cord injury (SCI) observed in humans can be replicated in adult rat models, which are widely used for laboratory studies of SCI rehabilitation. To ensure the effectiveness and efficiency of an SCI rat model, the minimal time spent performing the laminectomy procedure and the damage caused to the body are of great importance. We describe and evaluate the effectiveness and advantages of a laminectomy auxiliary device (LAD) for removing the rat vertebral lamina without injuring the spinal cord. The incision size, success rate, operation duration, body weight, BBB score, step detection, latency and amplitude of transcranial electrical motor-evoked potentials (tceMEPs), and serum MDA and SOD levels were recorded in 8 normal rats, 8 rats treated with traditional laminectomy and 8 rats treated with LAD laminectomy. Compared with traditional laminectomy, in our LAD, the surgical incision was smaller (approximately 2.2 and 1.3 cm, respectively), the success rate was higher (88.89% and 100%, respectively) and the duration shorter (14.644±1.617 and 4.821±0.668 minutes, respectively). Compared with normal rats, those treated with either laminectomy using LAD or the traditional method showed slower body weight gain and temporarily increased oxidative stress levels. However, there were no significant differences between these two groups. Our results show that laminectomy using this LAD provides three main advantages in rats: a high success rate, time savings, small incisions and reduced trauma. We believe this LAD can be used as an effective assistant tool for rodent laminectomy.

Cell transplantation of retinal ganglion cells derived from hESCs.

BACKGROUND: Glaucoma, the number one cause of irreversible blindness, is characterized by the loss of retinal ganglion cells (RGCs), which do not regenerate in humans or mammals after cell death. Cell transplantation provides an opportunity to restore vision in glaucoma, or other optic neuropathies. Since transplanting primary RGCs from deceased donor tissues may not be feasible, stem cell-derived RGCs could provide a plausible alternative source of donor cells for transplant. OBJECTIVE: We define a robust chemically defined protocol to differentiate human embryonic stem cells (hESCs) into RGC-like neurons. METHODS: Human embryonic stem cell lines (H7-A81 and H9) and induced pluripotent stem cell (iPSC) were used for RGC differentiation. RGC immaturity was measured by calcium imaging against muscimol. Cell markers were detected by immunofluorescence staining and qRT-PCR. RGC-like cells were intravitreally injected to rat eye, and co-stained with RBPMS and human nuclei markers. All experiments were conducted at least three times independently. Data were analyzed by ANOVA with Tukey's test with P value of <0.05 considered statistically significant. RESULTS: We detected retinal progenitor markers Rx and Pax6 after 15 days of differentiation, and the expression of markers for RGC-specific differentiation (Brn3a and Brn3b), maturation (synaptophysin) and neurite growth (ß-III-Tubulin) after an additional 15 days. We further examined the physiologic differentiation of these hESC-derived RGC-like progeny to those differentiated in vitro from primary rodent retinal progenitor cells (RPCs) with calcium imaging, and found that both populations demonstrate the immature RGC-like response to muscimol, a GABAA receptor agonist. By one week after transplant to the adult rat eye by intravitreal injection, the human RGC-like cells successfully migrated into the ganglion cell layer. CONCLUSIONS: Our protocol provides a novel, short, and cost-effective approach for RGC differentiation from hESCs, and may broaden the scope for cell replacement therapy in RGC-related optic neuropathies such as glaucoma.

Schnyder corneal dystrophy-associated UBIAD1 mutations cause corneal cholesterol accumulation by stabilizing HMG-CoA reductase.

Schnyder corneal dystrophy (SCD) is a rare genetic eye disease characterized by corneal opacification resulted from deposition of excess free cholesterol. UbiA prenyltransferase domain-containing protein-1 (UBIAD1) is an enzyme catalyzing biosynthesis of coenzyme Q10 and vitamin K2. More than 20 UBIAD1 mutations have been found to associate with human SCD. How these mutants contribute to SCD development is not fully understood. Here, we identified HMGCR as a binding partner of UBIAD1 using mass spectrometry. In contrast to the Golgi localization of wild-type UBIAD1, SCD-associated mutants mainly resided in the endoplasmic reticulum (ER) and competed with Insig-1 for HMGCR binding, thereby preventing HMGCR from degradation and increasing cholesterol biosynthesis. The heterozygous Ubiad1 G184R knock-in (Ubiad1G184R/+) mice expressed elevated levels of HMGCR protein in various tissues. The aged Ubiad1G184R/+ mice exhibited corneal opacification and free cholesterol accumulation, phenocopying clinical manifestations of SCD patients. In summary, these results demonstrate that SCD-associated mutations of UBIAD1 impair its ER-to-Golgi transportation and enhance its interaction with HMGCR. The stabilization of HMGCR by UBIAD1 increases cholesterol biosynthesis and eventually causes cholesterol accumulation in the cornea.

Opposing Effects of Growth and Differentiation Factors in Cell-Fate Specification.

Following ocular trauma or in diseases such as glaucoma, irreversible vision loss is due to the death of retinal ganglion cell (RGC) neurons. Although strategies to replace these lost cells include stem cell replacement therapy, few differentiated stem cells turn into RGC-like neurons. Understanding the regulatory mechanisms of RGC differentiation in vivo may improve outcomes of cell transplantation by directing the fate of undifferentiated cells toward mature RGCs. Here, we report a new mechanism by which growth and differentiation factor-15 (GDF-15), a ligand in the transforming growth factor-beta (TGF-ß) superfamily, strongly promotes RGC differentiation in the developing retina in vivo in rodent retinal progenitor cells (RPCs) and in human embryonic stem cells (hESCs). This effect is in direct contrast to the closely related ligand GDF-11, which suppresses RGC-fate specification. We find these opposing effects are due in part to GDF-15's ability to specifically suppress Smad-2, but not Smad-1, signaling induced by GDF-11, which can be recapitulated by pharmacologic or genetic blockade of Smad-2 in vivo to increase RGC specification. No other retinal cell types were affected by GDF-11 knockout, but a slight reduction in photoreceptor cells was observed by GDF-15 knockout in the developing retina in vivo. These data define a novel regulatory mechanism of GDFs' opposing effects and their relevance in RGC differentiation and suggest a potential approach for advancing ESC-to-RGC cell-based replacement therapies.

All-trans Retinoic Acid Regulates the Balance of Treg-Th17 Cells through ERK and P38 Signaling Pathway.

BACKGROUND: All-trans retinoic acid (ATRA) potentiates TGF-ß-dependent regulatory T cells (Treg) induction, while it inhibits pro-inflammatory interleukin-17-producing T helper cells (Th17) differentiation. Combined application of ATRA and TGF-ß may shift Treg-Th17 balance towards Treg. OBJECTIVE: To investigates the effect of ATRA on the regulation of Th17-Treg balance through ERK and p38 signaling pathway. METHODS: Mice naive CD4+T cells were isolated and co-cultured with 100 nmol/ml ATRA and 5 ng/ml TGF-ß. The effect of ATRA on the phosphorylation of ERK and P38 was evaluated. The induction of Treg and Th17 was investigated before and after the application of the inhibitor of ERK and P38. RESULTS: The expression of p-ERK1 and p-ERK2 increased significantly when the cells were incubated for 3 days with both TGF-ß and ATRA. The upregulated expression of p38 was found after incubation for 1 day. The inhibition of ERK prohibited Treg induction and promoted Th17 development. However, the inhibition of p38 only had inhibitory effect on Treg induction. CONCLUSIONS: ERK and p38 pathways participated in ATRA-activated Treg-Th17 balance adjustment.

Retinal Cell Fate Specification.

How are different neural cell types generated from progenitor cells? In 1990, Turner et al. used new lineage tracing techniques to show that different cells in the mammalian retina share their progenitor origin. The findings established a key step toward our understanding of how multipotent progenitor cells give rise to complex circuitry in the retina.

Induced Pluripotent Stem Cells Promote Retinal Ganglion Cell Survival After Transplant.

Purpose: The purpose of this study was to characterize whether induced pluripotent stem cells (iPSCs) affect survival of grafted retinal ganglion cells (RGCs) after transplantation. Methods: For in vitro studies, human iPSCs were either directly cocultured with mouse RGCs or plated in hanging inserts in RGC cultures for 1 week. For ex vivo studies, RGCs and iPSCs were seeded onto the inner surface of an adult rat retina explant and cultured for 1 week. For in vivo studies, RGCs and iPSCs were intravitreally coinjected into an adult rat eye 1 week before examining retinas by explant and immunostaining. Results: A dose-dependent increase in RGC survival was observed in RGC-iPSC direct cocultures, and RGC-iPSC indirect cocultures showed a similar RGC protective effect, but to a lesser extent than in direct coculture. Enhanced RGC survival was also identified in RGC-iPSC cotransplantations to adult retinas ex vivo and in vivo. In addition, RGCs with iPSC cotransplantation extended significantly longer neurites than RGC-only transplants. Conclusions: Human iPSCs promote transplanted RGC survival and neurite extension. This effect may be mediated at least partially through secretion of diffusible neuroprotective factors.

Effective robotic assistive pattern of treadmill training for spinal cord injury in a rat model.

The purpose of the present study was to establish an effective robotic assistive stepping pattern of body-weight-supported treadmill training based on a rat spinal cord injury (SCI) model and assess the effect by comparing this with another frequently used assistive stepping pattern. The recorded stepping patterns of both hind limbs of trained intact rats were edited to establish a 30-sec playback normal rat stepping pattern (NRSP). Step features (step length, step height, step number and swing duration), BBB scores, latencies, and amplitudes of the transcranial electrical motor-evoked potentials (tceMEPs) and neurofilament 200 (NF200) expression in the spinal cord lesion area during and after 3 weeks of body-weight-supported treadmill training (BWSTT) were compared in rats with spinal contusion receiving NRSP assistance (NRSPA) and those that received manual assistance (MA). Hind limb stepping performance among rats receiving NRSPA during BWSTT was greater than that among rats receiving MA in terms of longer step length, taller step height, and longer swing duration. Furthermore a higher BBB score was also indicated. The rats in the NRSPA group achieved superior results in the tceMEPs assessment and greater NF200 expression in the spinal cord lesion area compared with the rats in the MA group. These findings suggest NRSPA was an effective assistive pattern of treadmill training compared with MA based on the rat SCI model and this approach could be used as a new platform for animal experiments for better understanding the mechanisms of SCI rehabilitation.

Incidence and risk factors for post-penetrating keratoplasty glaucoma: A systematic review and meta-analysis.

OBJECTIVES: To establish the incidence and risk factors for post penetrating keratoplasty glaucoma (PKKG). METHODS: Studies published between 1947 and 2016 regarding penetrating keratoplasty (PK) were identified using an electronic search and reviewed. For search purpose, PKKG was defined as ocular hypertension (> 21mmHg) after PK. The incidence and risk factors of PKKG were extracted for all studies. Pooled incidence, odd ratios (ORs) and 95% confidence intervals (CIs) were calculated. RESULTS: Thirty studies reporting on 27146 patients were included in the analysis of the incidence and risk factors for PKKG. Exact PKKG definitions used in the literature could be classified in to three subgroups: I, ocular hypertension (> 21mmHg) after PK; II, I plus > 4 weeks medical treatment required; III, II plus treatment escalation among patients with preexisting glaucoma. Overall (Definition I) pooled incidence in all studies was 21.5% (95% CI 17.8%, 25.7%). The incidence varied according to different definitions. The highest incidence value was found when only studies using Goldmann tonometer were included (22.5%), while the lowest incidence was found when a strict definition was used and steroid-induced PPKG was excluded (12.1%). The incidence was higher in patients with preexisting glaucoma, bullous keratopathy (BK), aphakia, pseudophakia, failed graft, and surgical indication of trauma. A triple procedure (combined PK with extra capsular cataract extraction and intraocular lens implantation) was not identified as being associated with the increased risk for PKKG. CONCLUSIONS: The overall pooled incidence of PKKG was 21.5%, but it varied according to the criteria used to define the presence of PPKG. Strong risk factors for PKKG included preexisting glaucoma and aphakia, while modest predictors included pseudophakia, regrafting, and preoperative diagnosis like BK and trauma. There may not be sufficient evidence to identify a significant association between a triple procedure and PKKG.

Assessment of Bulbar Redness with a Newly Developed Keratograph.

PURPOSE: To determine the validity and reliability of the clinical assessment of bulbar redness (BR) using a newly developed corneal topographer. METHODS: This is a cross-sectional diagnostic evaluation study. The BR scores [Oculus Index (OI)] from 30 eyes of 26 patients with different degrees of conjunctival hyperemia were assessed and scored automatically using a new method: a keratograph equipped with scanning and scoring software. The values obtained via this system were correlated with three image-based comparative subjective scales: the Institute for Eye Research (IER), the Efron, and the Validated Bulbar Redness (VBR) grading scales. The IER and Efron scores were interpolated to 0.1 unit, and the VBR scores were interpolated to 1 unit. We also evaluated the repeatability of each method and the level of agreement between the OI score and the scores achieved using the three other image-based methods. RESULTS: There was a significant correlation between the OI score and the scores obtained with the IER (r = 0.921, p < 0.001), Efron (r = 0.958, p < 0.001), and VBR (r = 0.965, p < 0.001) scales. The intraobserver intraclass correlation coefficients were 0.947, 0.874, 0.810, and 0.920 for the OI, IER, Efron, and VBR, respectively, and the intraobserver coefficients of repeatability were 13.924, 16.111, 17.684, and 16.900, respectively. Furthermore, the interobserver intraclass correlation coefficients were 0.889, 0.880, 0.884, and 0.881 for the OI, IER, Efron, and VBR, respectively, and the interobserver coefficients of repeatability were 15.934, 16.366, 22.059, and 21.373, respectively. CONCLUSIONS: The OI is an objective and reliable method for scoring BR. Its reproducibility was the highest of all the four modalities. The keratograph is recommended, therefore, as a suitable alternative for BR assessment.

All-trans retinoid acid promotes allogeneic corneal graft survival in mice by regulating Treg-Th17 balance in the presence of TGF-ß.

BACKGROUND: All-trans retinoid acid (ATRA) has been proven to skew Regulatory T cell-T helper 17 cell (Treg-Th17) balance toward Treg in vitro, favoring graft acceptance. However, its in vivo effect after solid organ transplantation is under investigation. RESULTS: BALB/c mice were given orthotopic corneal grafts from C57BL/6 donors, and recipient mice were administered with ATRA, TGF-ß, and the combination of both agents for 8 weeks after surgery. We found that a mixed treatment of ATRA and TGF-ß significantly promoted graft survival. Moreover, with the presence of TGF-ß, ATRA upregulated CD4(+)CD25(+)Foxp3(+)Treg cells and suppressed Th17 cells in the blood, spleen and draining lymph nodes of recipient mice, as well as enhanced the Foxp3 expression and inhibited the RORγt expression in grafts and peripheral blood mononuclear cells (PBMCs). Simultaneously, increased number of Foxp3+ cells and decreased number of IL-17+ cells in conjunctiva were found in recipients with mixed treatment, along with reduced IL-17 level in serum and aqueous humor and increased IL-10 level in aqueous humor. Tregs isolated from recipient mice treated with ATRA + TGF-ß presented the strongest suppressive activity in vitro. CONCLUSIONS: Combined application of ATRA and TGF-ß may shift the Th17-Treg balance toward Tregs, hence facilitating the induction of immunological tolerance after allogenic corneal transplantation and representing a potential therapeutic approach in the treatment of posttransplant rejection.

[The study of corneal epithelial change in keratoconic eyes with Fourier-domain optical coherence tomographic pachymetry].

OBJECTIVE: To compare the corneal epithelial thickness profile in keratoconic and normal eyes with advanced Fourier-domain optical coherence tomography (OCT) and to evaluate the diagnostic efficiency of pachymetry parameters. METHODS: Cross-sectional observational study. Twenty-five consecutive keratoconus patients (34 eyes) and 41 normal volunteers (41 eyes) in Eye & ENT Hospital of Fudan University between June 1 and July 31, 2013 were enrolled.Observations of corneal epithelial thickness and corneal thickness profiles were taken from all subjects. The epithelial pachymetry parameters were superior, inferior, inferior-superior (I-S), minimum (Min), maximum (Max), Min-Max, and central. The corneal pachymetry parameters were inferotemporal-superonasal (IT-SN), I-S, Min, Min-median, Min-Max, and central.Statistical analysis methods included Chi-square test, independent two-sample t-test, and Wilcoxon rank sum test. The diagnostic efficiency of all parameters were analyzed by receiver operating characteristic (ROC) curves and Youden index. RESULTS: The inferior, I-S, Min, Min-Max, and central epithelial thickness averages were (50.5 ± 4.8) µm, (-2.7 ± 7.5) µm, (35.9 ± 10.7) µm, (-33.6 ± 18.9) µm, and (50.1 ± 6.8) µm in keratoconic eyes.In normal eyes, they were (54.8 ± 2.5) µm, (0.9 ± 1.3) µm, (51.4 ± 2.6) µm, (-5.7 ± 1.8) µm, and (54.7 ± 2.2) µm. Among the epithelial pachymetry parameters, Min-Max and Min showed high diagnostic efficiency for keratoconus (AUC = 0.967, 0.968; Youden index = 0.882, 0.829); inferior, I-S, and central parameters showed moderate efficiency (AUC = 0.794, 0.728, 0.715; Youden index = 0.461, 0.564, 0.565). The IT-SN, I-S, Min, Min-median, Min-Max, and central corneal thickness averages were (-82.8 ± 65.2) µm, (-72.5 ± 55.6) µm, (401.2 ± 83.8) µm, (-83.4 ± 48.2) µm, (-169.7 ± 99.4) µm, and (453.4 ± 58.6) µm in keratoconic eyes.In normal eyes, these parameters were respectively (-23.7 ± 18.5) µm, (-20.5 ± 19.8) µm, (525.4 ± 28.1) µm, (-23.1 ± 4.8) µm, (-59.2 ± 23.3) µm, and (533.4 ± 28.8) µm. Min, Min-median, and Min-Max had preferable diagnostic efficiency (AUC = 0.952, 0.938, 0.924; Youden index = 0.784, 0.829, 0.721); central, IT-SN, and I-S parameters showed moderate efficiency (AUC = 0.891, 0.877, 0.862; Youden index = 0.643, 0.707, 0.580). CONCLUSIONS: In observation with Fourier-domain OCT, keratoconus was characterized by central and inferior epithelial thinning, increased epithelial thickness variation, central cornea thinning, increase in asymmetry, and gaps between corneal IT-SN and I-S thickness.Fourier-domain OCT was confirmed to have high speed, high resolution, and noninvasion. The pachymetry parameters showed specific strengths in diagnosis of keratoconus, especially epithelial pachymetry parameters of Min and Min-Max, and corneal pachymetry parameters of Min, Min-median and Min-Max.

Production and in vitro antioxidant activity of exopolysaccharide by a mutant, Cordyceps militaris SU5-08.

Cordyceps militaris SU5-08 was derived from an initial strain (C. militaris SU5) by ultraviolet mutagenesis of protoplasts, and the extraction parameters for C. militaris SU5-08 exopolysaccharide (EPS) produced during submerged culture were optimized. The extraction rate of EPS was 1919.16±165.27 mg/l, which was 120.38±11.36% higher than that of C. militaris SU5. The in vitro scavenging effects of EPS of C. militaris SU5-08 on hydroxyl, superoxide anion and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at a dosage of 5 g/l were 63.64±3.52%, 75.27±5.16%, and 6.46±5.03%, respectively. The reducing power of EPS of C. militaris SU5-08 was 0.21±0.01. The results suggest that the EPS of C. militaris SU5-08 can be used as a potential antioxidant which enhances adaptive immune responses.