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The fabrication of MCL and the mechanism of MCL-mediated antitumor effects against melanoma.

Autologous tumor cells and cell-derived secretions (CDS) can induce antitumor immune responses. The conditions in which cells are cultured and treated impact CDS, and cellular insults alter their composition and function. In this study, we generated CDS from tumor cells exposed to normal culture conditions, hypoxia, cisplatin, radiotherapy, photodynamic therapy, or hypochlorous acid (HOCl). In vitro HOCl-CDS showed the strongest stimulatory effects on dendritic cells and macrophages compared to CDS generated by hypoxia, cisplatin, radiotherapy or photodynamic therapy. To improve HOCl-CDS activity at the tumor site, we loaded HOCl-CDS into a melittin-encapsulated hydrogel scaffold. When injected intratumorally, the HOCl-CDS hydrogel promoted tumor cell death, cytotoxic T lymphocyte infiltration, and tumor-associated macrophage reprogramming towards an M1 phenotype. The hydrogel inhibited tumor growth and prolonged the survival of mice bearing B16–F10 melanoma. Furthermore, hydrogel-delivered HOCl-CDS augmented the antitumor effects of immune checkpoint blockade. These results underscore the importance of the CDS generation method and delivery approach for improving cancer immunotherapy., Graphical abstract Scheme 1. Schematic diagram for generating the HOCl-CDS and HOCl-CDS hydrogel for potentiated cancer immunotherapy.Image 1, Highlights • HOCl-treated tumor cell-derived secretions (HOCl-CDS) is a robust immune-stimulator on dendritic cells and macrophages. • A multifunctional HOCl-CDS hydrogel was developed by loading HOCl-CDS into a melittin-encapsulated hydrogel scaffold. • HOCl-CDS hydrogel promoted tumor cell death, cytotoxic T lymphocyte infiltration and M1-TAM polarization in mice. • HOCl-CDS hydrogel synergistically augmented the therapeutic effect of anti-PD-1 and further potentiated cancer immunotherapy.

Diagnosis of the Graves’ ophthalmology remains a significant challenge. We identified between Graves’ ophthalmology tissues and healthy controls by using laser-induced breakdown spectroscopy (LIBS) combined with machine learning method. In this work, the paraffin-embedded samples of the Graves’ ophthalmology were prepared for LIBS spectra acquisition. The metallic elements (Na, K, Al, Ca), non-metallic element (O) and molecular bands ((C-N), (C-O)) were selected for diagnosing Graves’ ophthalmology. The selected spectral lines were inputted into the supervised classification methods including linear discriminant analysis (LDA), support vector machine (SVM), k-nearest neighbor (kNN), and generalized regression neural network (GRNN), respectively. The results showed that the predicted accuracy rates of LDA, SVM, kNN, GRNN were 76.33%, 96.28%, 96.56%, and 96.33%, respectively. The sensitivity of four models were 75.89%, 93.78%, 96.78%, and 96.67%, respectively. The specificity of four models were 76.78%, 98.78%, 96.33%, and 96.00%, respectively. This demonstrated that LIBS assisted with a nonlinear model can be used to identify Graves’ ophthalmopathy with a higher rate of accuracy. The kNN had the best performance by comparing the three nonlinear models. Therefore, LIBS combined with machine learning method can be an effective way to discriminate Graves’ ophthalmology.

Anticancer peptides (ACPs) have gained significant attention in the past few years. Most ACPs only act toward intracellular targets. However, their low membrane penetrability often limits their anticancer efficacy. Here we developed a novel melittin-RADA28 (MR) hydrogel, composed of RADA28 and melittin, through a peptide fusion method in order to promote the membrane permeability of tumor cells with the membrane-disrupting ability of melittin. As a proof of concept, we loaded the MR hydrogel with a therapeutic peptide, KLA (KLAKLAKKLAKLAK), to show the enhanced delivery efficiency of the hydrogel. Our results demonstrated that the formed melittin-RADA28-KLA peptide (MRP) hydrogel has a nanofiber structure, sustained release profile, and attenuated hemolysis effects. Compared with free KLA, the MRP hydrogel markedly increased the cellular accumulation of KLA, produced the highest ratio of the depolarized mitochondrial membrane, and decreased cell viability in vitro. Following peritumoral injection, the MRP hydrogel treatment suppressed CT26 tumor growth by more than 85%, compared to controls. In summary, we provide a facile and efficient strategy to enhance the delivery of impermeable peptides to improve their therapeutic efficiency.

Background Graft-versus-host disease (GVHD) is the most common complication after bone marrow allogeneic hematopoietic stem cell transplantation (allo-HSCT). The incidence of posterior segment complications was significantly lower than that of ocular surface lesions. Up to now, there has been no report about optic neuropathy associated with GVHD. Case presentation A 23-year-old man presented with visual acuity decline after allo-HSCT for B-acute lymphoblastic leukemia (B-ALL). Red rashes were found all over the body simultaneously. Visual field examination revealed central scotomas in both eyes. Visual evoked potential showed prolonged P100 latency and decreased P100 amplitude in both eyes. Other ocular examinations showed no obvious abnormality except for blunt pupillary light reflex. The minimal residual disease test was negative after transplantation, and no obvious abnormalities were found in optic nerve and brain by magnetic resonance imaging (MRI). After the multi-disciplinary consultation, the rashes and optic neuropathy were considered GVHD probably. As for the treatment, methylprednisolone and Ruxolitinib were suggested, supported by adjunctive neurotrophic therapy. Two months later, the rashes gradually subsided. However, the visual acuity was not significantly improved at latest follow-up. Conclusions The present case report demonstrated GVHD probably associated with optic neuropathy. Although extremely rare, optic nerve should be considered as a potential target of ocular GVHD, which could expand the dimensions of GVHD.

To explore visual dysfunction in Graves' orbitopathy (GO) objectively by analyzing chromatic visual evoked potentials (cVEP) and evaluate its diagnostic efficiency for dysthyroid optic neuropathy (DON).In this cross-sectional study, we analyzed pattern-reversal VEP (pVEP), red-green (R-G) and blue-yellow (B-Y) cVEP in 93 subjects (21 with DON, Group A, 30 with GO, Group B, and 42 healthy controls, Group C) at Wuhan Union Hospital, China.Compared with Group C, the amplitudes of B-Y cVEP were significantly lower in Group B, whereas all amplitudes of cVEP, latencies and amplitudes of pVEP in Group A were significantly impaired. In addition, the pVEP latency at 60 arcmin (60'), pVEP amplitudes and R-G cVEP amplitudes were significantly different between Group A and B. Moreover, 60'cVEP R-G negative-positive (N-P) amplitude was correlated with crowding index (P = 0.001), the average thickness of ganglion cell layer and inner plexiform layer (P = 0.004). Furthermore, combination of 60'cVEP R-G amplitude and 60'pVEP P100 latency had better diagnostic efficiency than each single parameter, with optimal cut-off values of 14.20 μV and 110.65 ms, respectively.GO may induce electrophysiological changes. The presence of B-Y cVEP anomalies in moderate to severe GO patients may be an early sign of preclinical DON. A decline in 60'cVEP R-G amplitude is associated with apical crowding and thinner inner intra-retinal layers. The combination of 60'cVEP R-G N-P amplitude and 60'pVEP latency can be a useful diagnostic index for DON.

To determine the prognostic value of optical coherence tomography (OCT) measurement of the peripapillary retinal nerve fiber layer (RNFL) thickness in visual recovery after orbital decompression of patients with dysthyroid optic neuropathy (DON). A total of 52 eyes of 37 patients who underwent orbital decompression for DON between 2013 and 2019 were retrospectively reviewed. We examined peripapillary RNFL thickness, best-corrected visual acuity (BCVA), visual field (VF) for mean deviation (MD) and pattern standard deviation (PSD), and pattern-reversed visual evoked potential (PVEP) for P100 latency and amplitude before and after surgery. Black and white checkerboard square sizes of PVEP were 15 and 60 arcmin (arcminute and minute of angle). Changes in RNFL overall thickness and by quadrant and interocular differences were evaluated and studied regarding changes in BCVA, VF and PVEP. There was a significant improvement in BCVA, VF, and PVEP, whereas a dramatic reduction in RNFL thickness of all DON patients in global average, temporal, superior, and inferior quadrants (P = 0.005, P = 0.024, P = 0.016, and P = 0.001, respectively) after decompression surgery, except for nasal quadrant (P = 0.057). The preoperative RNFL thickness in each quadrant was negatively correlated with postoperative changes of BCVA and PSD and positively correlated with changes of MD and P100 amplitude at 60 arcmin (all P

The cornea is one of the major refractive eye components and could be easily injured. An ineffective healing of corneal stromal wound may cause fibrosis and even loss of vision. Therefore, it is pivotal to prevent corneal fibrosis after injury. In this study, a poly (ε-caprolactone) (PCL) microfibrous scaffold infused with rat tail collagen type I was fabricated to obtain a 3D composite material. Physical and biological properties of PCL/collagen scaffold were evaluated, the effect of PCL/collagen scaffold on the proliferation and differentiation of limbal stromal stem cells (LSSCs) were detected in vitro, the differentiation of keratocytes as well as the expression and arrangement of extracellular matrix (ECM) influenced by PCL/collagen scaffold were investigated in vivo. RNA-sequencing on normal and injured corneas was carried out to find out the differential enriched pathways and gene expression. We discovered that the PCL/collagen scaffold simulated the stromal structure with properties that were most similar to the native cornea, the PCL/collagen scaffold exhibited good mechanical and biological properties. We also observed that the PCL/collagen scaffold reduced keratocyte differentiation. Injured corneas treated with PCL/collagen scaffold exhibited more regular collagen distribution and less fibroblasts and myofibroblasts distribution. By RNA-sequencing, we observed that in injured group, ECM-related pathway was enriched and several ECM-related genes were up-regulated. This study provides evidence that application of PCL/collagen scaffold could be a new therapeutic strategy for corneal injury.

Correction for ‘Melittin-encapsulating peptide hydrogels for enhanced delivery of impermeable anticancer peptides’ by Jue-Ping Feng et al., Biomater. Sci., 2020, 8, 4559–4569, DOI: 10.1039/C9BM02080B.

BACKGROUND Calcium sensing receptor (CaSR) is widely expressed in many tissues of the body, but it is rarely reported to be expressed in the eyes. This research explored the expression and distribution of CaSR in eye tissues of normal and diabetic rats. MATERIAL AND METHODS Thirty male Sprague Dawley (SD) rats were randomly divided into a diabetic and a normal group. Diabetes mellitus (DM) models were successfully established by intraperitoneal injection of streptozotocin. The expression and distribution of CaSR in the rat eyeballs were detected by immunohistochemistry. Quantitative RT-PCR and western blotting were used to detect the presence of CaSR in normal and diabetic rats. RESULTS CaSR was detected in the cornea, lens epithelium, and retina. CaSR was expressed the most in the cornea, followed by the lens epithelium, and the retina (p

To compare visual function of 2-wall (medial and lateral) versus 3-wall (medial, lateral, and inferior) orbital decompression in patients with dysthyroid optic neuropathy (DON). A total of 52 eyes of 37 patients underwent orbital decompression for DON between 2013 and 2019 were retrospectively reviewed. Two- or 3-wall decompression was performed in 31 eyes of 23 patients and 21 eyes of 14 patients, respectively. We examined best-corrected visual acuity (BCVA), visual field mean deviation (MD) and pattern standard deviation (PSD), pattern-reversed visual evoked potential (PVEP) for P100 latency and amplitude at 60 and 15 arcmin stimulation checkerboard size, as well as proptosis using Hertel exophthalmometry. Whether 2-wall or 3-wall decompression, all parameters of visual function were improved after surgery (all P .05). Proptosis reduction was higher after 3-wall decompression (P = .011). Mean increase in P100 amplitude after 3-wall decompression was statistically higher than that of after 2-wall decompression at 60 and 15 arcmin (P = .045 and .020, respectively), while the mean decrease in P100 latency was similar between the groups (P = .821 and .655, respectively). Six patients (66.67%) had persistent postoperative diplopia and 1 patient (20%) had new-onset diplopia in 3-wall decompression group, which were higher than in 2-wall decompression group (46.15% persistent postoperative diplopia and no new-onset diplopia). Both 2-wall and 3-wall decompression can effectively improve visual function of patients with DON. Three-wall decompression provides better improvement in P100 amplitude and proptosis, however new-onset diplopia is more common with this surgical technique.

Doxorubicin (DOX) is known to induce apoptosis and necrosis in healthy tissue resulting in unwanted toxicities. To improve the ability of DOX to more specifically target tumors and minimize undesirable side effects, conjugation of DOX with epidermal growth factor receptor (EGFR)--binding peptide (DOX-EBP) has been developed to deliver DOX to EGFR-overexpressing neoplastic cells. Here, we investigated whether DOX-EBP was able to reduce toxicity and enhance anticancer efficacy in vivo through receptor-mediated targeted delivery system. Nude mice were treated with DOX or DOX-EBP to estimate general toxicity, normal tissue damage, biodistribution, and antitumor efficacy. In addition, the expression levels of EGFR in tumor tissues and normal organs were investigated by Western blotting, and their mRNA expression was analyzed by reverse transcription PCR. This study demonstrated that DOX-EBP was able to effectively decrease the distribution of DOX in normal tissues without EGFR overexpressing and reduce DOX-induced toxicity. On the other hand, the research also confirmed that DOX-EBP was able to preferentially accumulate DOX in EGFR-overexpressing tumor tissues and showed the enhanced anticancer efficacy over free DOX. DOX-EBP could be used for receptor-targeted chemotherapy with less toxicity and greater efficacy of tumor cells overexpressing EGFR. DOX-EBP conjugate is a good therapeutic agent for cancer treatment.

To describe the clinical and genetic findings in a Chinese family with three sibs diagnosed with Usher syndrome type IIC.Four members received ophthalmic and otologic tests to ascertain the clinical characteristics. According to the clinical phenotype, we focused attention on a total of 658 genes associated with them. We screened the possible pathogenic mutation sites, used Sanger to exclude the false positive and verified whether there were co-segregated among the family members.Typical fundus features found in the proband supported the diagnosis of retinitis pigmentosa (RP). Audiometric test indicated moderate to severe sensorineural hearing impairment while the vestibular function was normal. Whole-exome sequencing identified the presence of two novel compound heterozygous mutations in ADGRV1, a known gene responsible for Usher syndrome type IIC. Mutationc.15008delG/p.Gly5003AlafsTer13 was inherited from the mother while c.18383_18386dupACAG/p.His6130GlnfsTer84 was inherited from the father, and they were co-segregated with the disease phenotype in the family.The mutations found in our study not only broaden the mutation spectrum of ADGRV1, but also provide assistances for future genetic diagnosis and treatment for Usher syndrome patients.

Immunosuppressive tumor microenvironments (TMEs) create tremendous obstacles for an effective cancer therapy. Herein, we developed a melittin-RADA32 hybrid peptide hydrogel loaded with doxorubicin (DOX) for a potent chemoimmunotherapy against melanoma through the active regulation of TMEs. The formed melittin-RADA32-DOX (MRD) hydrogel has an interweaving nanofiber structure and exhibits excellent biocompatibility, controlled drug release properties both in vitro and in vivo, and an enhanced killing effect to melanoma cells. A single-dose injection of MRD hydrogel retarded the growth of primary melanoma tumors by more than 95% due to loaded melittin and DOX, with concomitant recruitment of activated natural killer cells in the tumors. Furthermore, MRD hydrogel can activate dendritic cells of draining lymph nodes, specifically deplete M2-like tumor-associated macrophages (TAMs), and produce active, cytotoxic T cells to further defend the cells against remaining tumors, providing potent anticancer efficacy a...

The objectives of this study were to investigate the distributions of abnormally expressed optineurin (OPTN) proteins in retinal ganglion cells (RGC5s) of transgenic rats and their effects on subcellular morphological structures. Green fluorescent protein labeled EGFP wild-type (OPTN(WT)), E50K mutant type (OPTN(E50K)), and OPTN siRNA (si-OPTN) eukaryotic expression plasmids were constructed and transfected into RGC5s. Intracellular structures were labeled with organelle specific fluorescent dyes. Construct localization and cell morphologies were visualized by confocal fluorescence microscopy. OPTN(WT) was observed to be distributed as fine punctate fluorescent particles in the cytoplasm around the nucleus, along with exhibiting nuclear expression. OPTNE50K exhibited similar distribution but with non-uniform fluorescence particle size. si-OPTN distribution was similar to that of EGFP: uniform across the cytoplasm and nucleus. Compared with the negative control group, OPTN(WT), and OPTN(E50K) and to a lesser degree pEGFP-transfected cells exhibited fracture and loss of myofilament proteins and mitochondrial swelling and cytoplasmic accumulation, along with abnormal lysosomal distribution and increased volume, and Golgi fragmentation. However, si- OPTN transfected cells exhibited no significant damage. Therefore, we demonstrated that the E50K mutation disrupts the uniformity of OPTN protein distribution upon exogenous overexpression. Furthermore, these results suggested that si-OPTN transfection, and thus potentially OPTN knockdown, did not impact subcellular morphology of RGC5 cells, whereas transfection, especially when combined with wild-type or mutant OPTN expression, led to substantial abnormalities in subcellular morphological structures. These findings lay a foundation for further research into the function of the OPTN protein.

Diabetic retinopathy (DR) is a major microvascular complication in diabetics, and its mechanism is not fully understood. Toll-like receptor 4 (TLR4) plays a pivotal role in the maintenance of the inflammatory state during DR, and the deletion of TLR4 eventually alleviates the diabetic inflammatory state. To further elucidate the mechanism of DR, we used bone marrow transplantation to establish reciprocal chimeric animals of TLR4 mutant mice and TLR4 WT mice combined with diabetes mellitus (DM) induction by streptozotocin (STZ) treatment to identify the role of TLR4 in different cell types in the development of the proinflammatory state during DR. TLR4 mutation did not block the occurrence of high blood glucose after STZ injection compared with WT mice but did alleviate the progression of DR and alter the expression of the small vessel proliferation-related genes, vascular endothelial growth factor (VEGF), and hypoxia inducible factor-1α(HIF-1α). Grafting bone marrow-derived cells from TLR4 WT mice into TLR4 mutant mice increased the levels of TNF-α, IL-1β, and MIP-2 and increased the damage to the retina. Similarly, VEGF and HIF-1αexpression were restored by the bone marrow transplantation. These findings identify an essential role for TLR4 in bone marrow-derived cells contributing to the progression of DR.

In recent years, deep learning has made great progress in computer vision, and shown a good application prospect in reading medical images. TAO (thyroid-associated ophthalmopathy) is one of the most common orbital diseases in adults is autoimmune disease, the exact pathogenesis is not clear. In hospitals, CT images are usually used for disease analysis and the distortion of ocular rectus muscles is one of the main causes of TAO, thus the separation of the ocular rectus muscle is of great practical significance. However, the artificial segmentation of the ocular rectus muscle is a time-consuming task and relies heavily on the experience of the operato. In this paper, we improved the traditional U-net with GoogLeNet inception module and apply it on the segmentation of ocular rectus muscles. The experimental results show that the segmentation results of our method are more accurate and efficient than the traditional algorithm, and it is helpful for doctors to diagnose patients’ diseases.

We investigate photonic crystal fibers (PCFs) with nanosize airholes (NAHs) in the core for the blue extension of supercontinuum generation. The principle behind the design is to enhance the evanescent wave in the IR part of the supercontinuum. When the zero-dispersion wavelength is fixed around 1 μm, the infrared wavelength of 2500 nm of the proposed fiber is able to group-velocity match to the short-wavelength of 403 nm, which is about 60 nm shorter than that of a conventional high-Δ PCF. Simultaneously, the nonlinearity is enhanced about three times. The dependence of PCF characteristics on NAHs is also discussed. Simulated results confirm the possibility of increasing the blueshift of the generated supercontinua in designed PCFs.

This study aimed to modify the mixed and purified culture of rat retinal ganglion cells (RGCs) in vitro. The retinae of 1-3 day old Sprague-Dawley (SD) rats were separated bluntly into two layers: inner layer and outer layer, under a surgical microscope. Retinal cells isolated from different layers (inner layer, outer layer and whole retinal tissue) by using enzyme dissociation method were cultured in F12/DMEM medium containing 15% FBS. After 3-day culture, the RGCs in the retinal cells obtained from mixed culture of inner, outer, and whole retinal tissue were identified by immunocytochemical staining of Thy-1.1, and the rate of RGCs to retinal cells (RGCs%) was calculated. Two monoclonal antibodies, anti-macrophages/granulocytes (OX-41) against rat macrophage and antibody against rat Thy-1.1 (OX-7), were used to purify RGCs by either a conventional or modified two-stepped immunopanning procedure (purification in situ). Purified RGCs were seeded at different cell density and cultured in F12/DMEM medium containing 15% FBS. Immunocytochemical staining for Thy-1.1, MTT, and PI-Hoechst33342 fluorescence imaging were used to identify the purity and the viability of RGCs in purified culture of RGCs. The results showed: (1) Immunocytochemistry of different retinal tissue layers culture revealed that the RGCs% was (19.9 ± 1.2)%, (0.5 ± 0.2)%, and (6.2 ± 1.7)% respectively in the mixed culture of inner, outer, and whole retinal tissue, with differences being significant (P0.05); (2) fluorescent double staining of Hoechst33342 and PI indicated that with the same RGCs%, RGCs obtained from purification in situ grew well with more neurite outgrowth than those by the conventional two-stepped immunopanning method; (3) the viability of purified RGCs seeded at high density was increased and the cells developed complex intercellular networks. The viability of RGCs was declined with the decreasing seeding density, and most cells presented round or oval in shape with thin neurites. It was concluded that: (1) RGCs% in the inner layer retina was higher than that in the outer layer retina; (2) RGCs obtained by in situ purification had more neurite outgrowth and lower mortality than those by conventional two-stepped immunopanning procedure; (3) the viability of purified RGCs could be increased by increasing cell seeding density to some extent.

Femtosecond laser technology, used as a minimally invasive tool in intrastromal refractive surgery, may also have potential as a useful instrument for glaucoma filtration surgery. The purpose of the present study was to evaluate the feasibility of minimally invasive laser sclerostomy by femtosecond laser photodisruption and seek the appropriate patterns of laser ablation and relevant laser parameters. A femtosecond laser (800 nm/50 fs/1 kHz), focused by a 0.1 numerical aperture (NA) objective lens, with different pulse energies and exposure times was applied to ablate hydrated rabbit sclera in vitro. The irradiated samples were examined by scanning electron microscopy (SEM). By moving a three-dimensional, computer-controlled translation stage to which the sample was attached, the femtosecond laser could produce three types of ablation patterns, including linear ablation, cylindrical aperture and rectangular cavity. With pulse energies ranging from 37.5 to 150 μJ, the linear lesions were consistently observed at the inner surface of sclera, whereas it failed to make any photodisruption if pulse energy was below the threshold value of 31.25 μJ, with the corresponding threshold intensity of 4.06×1014 W/cm2. The depths of the linear lesions increased linearly with both pulse energy (37.5–150 μJ) and exposure time (0.1–0.4 s). Histological examination showed the incisions produced by femtosecond laser photodisruption had precise geometry and the edges were sharp and smooth, with no evidence of collateral damage to the surrounding tissue. Our results predict the potential application of femtosecond laser pulses in minimally invasive laser sclerostomy for glaucoma treatment.

To explore the possibility of photodisruption in rabbit sclera by femtosecond (fs) laser and seek appropriate incision techniques and relevant parameters, a fs laser (800 nm/50 fs) with different pulse energies was applied to irradiate rabbit sclera in vitro. By moving a computer-controlled three-axis translation stage to which the sample was attached, the laser achieved three types of incisions: transscleral channel, snake pattern and linear cut. The irradiated samples were observed by light microscopy and scanning electron microscopy (SEM). In comparison with fs laser, Nd:YAG was used as control. The experimental results show that through an objective lens with numerical aperture (NA) of 0.2, the fs laser with power intensity larger than 955 TW/cm2 and pulse energy ranging from 37.5–125 μJ, cutting depths from 30–70 μm may be achieved after linearly scanning in sclera at a translation speed of 0.1 mm/s. However, it failed to make any photodisruption when the power intensity was below 796 TW/cm2 or the pulse energy was less than 31.25 μJ under the same condition. Compared with the Nd:YAG laser, the inner wall of the channel was smoother and the damage to the surrounding tissues was slight with the fs laser. The high precision of intrascleral photodisruption and minimal damage to surrounding tissues by a fs laser display its potential application in the treatment of glaucoma.

The OPTN gene is thought to be associated with certain types of glaucoma and the function of the protein for which it encodes, optineurin, has been extensively researched, but with contradictory results. We explored the effects of abnormal optineurin expression on the survival of the rat retinal ganglion cell line RGC-5. Plasmids expressing wild-type (WT) or E50K mutant optineurin, or OPTN-specific double-hairpin small interfering RNA (si-RNA), were transfected into RGC-5 cells. The effects on cell survival were monitored by observation of cell morphology and propidium iodide and Hoechst 33342 fluorescent staining, while expression of optineurin was visualized by fluorescence microscopy. Abnormal optineurin expression influenced the survival of RGCs in vitro, as apoptosis was induced by increased WT and E50K mutant optineurin, while a reduction in apoptosis was observed in cells transfected with OPTN-siRNA. Similar results were also observed in transfected cells treated with apoptotic stimuli. Overexpression of WT and mutant E50K protein resulted in greater cell death, while downregulation decreased RGC-5 apoptosis.

AimsTo explore the role of the suppressor of cytokine signalling 3 (SOCS3) gene in Graves’ ophthalmopathy (GO) patients.MethodsA case–control study was conducted in a Chinese Han population by recruiting 114 Graves’ disease (GD) patients with GO and 156 GD patients without GO. We determined SOCS3 mRNA and protein levels in Epstein–Barr virus-transformed lymphoblastoid cell lines (EBV-LCLs) from peripheral blood mononuclear cells (PBMCs) by quantitative real-time (QRT)-PCR analysis and western blot analysis. We also genotyped five single nucleotide polymorphisms (SNPs) in the SOCS3 locus (SOCS3 rs12952093, rs4969170, rs4969168, rs4969169 and rs2280148) in all 270 GD patients using ligase detection reaction and multiplex PCR analyses. QRT-PCR and western blot assays were then performed to compare SOCS3 mRNA and protein levels between the rs4969170 AA and GG genotype groups from 20 GO patients.ResultsBasal SOCS3 mRNA and protein expression levels were significantly increased in patients with GO (pConclusionsPatients with GD who carry the AA genotype of the rs4969170 SNP in SOCS3 are more susceptible to the development of GO.

The method of gene transfer into corneal endothelium was investigated to provide a foundation for the study of TGF-beta 1 gene transfer to inhibit corneal graft rejection. Two days after direct injection of pMAM TGF-beta 1 mediated by liposome into the anterior chamber of rabbits, one half of corneas were made into paraffin slides and the endothelial layer was carefully torn from the other half to make a single layer slide of endothelia. By means of immunohistochemical technique, the plasmid pMAM TGF-beta 1 expression product TGF-beta 1 in the endothelia was detected. Specific TGF-beta 1 expression was positive in the endothelia on both the paraffin slide and the single layer slide. The results showed that by direct injection into the anterior chamber, foreign plasmid DNA could be transferred into the endothelia and its expression was obtained. This may provide a foundation for further study on TGF-beta 1 participating in local induction of corneal immune tolerance.

Of the 168 eyes undergoing silicone oil removal from 1993 to 2000, 34 developed retinal detachment after the removal of silicone oil. The management and the results of the 34 eyes were reviewed. Surgical procedures included pars plana vitrectomy, encircling band, cryocoagulation, endotamponade. Five of the 34 eyes had a second retinal detachment after these operations. One of these 5 eyes had a third retinal detachment. Final retinal reattachment was achieved in all eyes. Visual acuity decreased in 16 of the 34 eyes with retinal detachment, and 18 of the 34 eyes had a visual acuity of less than 0.02. These results show that surgery for retinal detachment after silicone oil removal yields a high rate of anatomic success, while the visual outcome is poor for a substantial number of patients.