Rapamycin improves Graves' orbitopathy by suppressing CD4+ cytotoxic T lymphocytes.

CD4+ cytotoxic T lymphocytes (CTLs) were recently implicated in immune-mediated inflammation and fibrosis progression of Graves' orbitopathy (GO). However, little is known about therapeutic targeting of CD4+ CTLs. Herein, we studied the effect of rapamycin, an approved mTOR complex 1 (mTORC1) inhibitor, in a GO mouse model, in vitro, and in patients with refractory GO. In the adenovirus-induced model, rapamycin significantly decreased the incidence of GO. This was accompanied by the reduction of both CD4+ CTLs and the reduction of orbital inflammation, adipogenesis, and fibrosis. CD4+ CTLs from patients with active GO showed upregulation of the mTOR pathway, while rapamycin decreased their proportions and cytotoxic function. Low-dose rapamycin treatment substantially improved diplopia and the clinical activity score in steroid-refractory patients with GO. Single-cell RNA-Seq revealed that eye motility improvement was closely related to suppression of inflammation and chemotaxis in CD4+ CTLs. In conclusion, rapamycin is a promising treatment for CD4+ CTL-mediated inflammation and fibrosis in GO.

iTRAQ-based comparative proteomic analysis provides insights into somatic embryogenesis in Gossypium hirsutum L.

KEY MESSAGE: iTRAQ based proteomic identified key proteins and provided new insights into the molecular mechanisms underlying somatic embryogenesis in cotton. Somatic embryogenesis, which involves cell dedifferentiation and redifferentiation, has been used as a model system for understanding molecular events of plant embryo development in vitro. In this study, we performed comparative proteomics analysis using samples of non-embryogenic callus (NEC), embryogenic callus (EC) and somatic embryo (SE) using the isobaric tags for relative and absolute quantitation (iTRAQ) technology. In total, 5892 proteins were identified amongst the three samples. The majority of these proteins (93.4%) were found to have catalytic activity, binding activity, transporter activity or structural molecular activity. Of these proteins, 1024 and 858 were differentially expressed in NEC versus EC and EC versus SE, respectively. Compared to NEC, EC had 452 and 572 down- and up-regulated proteins, respectively, and compared to EC, SE had 647 and 221 down- and up-regulated proteins, respectively. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis indicated that genetic information transmission, plant hormone transduction, glycolysis, fatty acid biosynthesis and metabolism, galactose metabolism were the top pathways involved in somatic embryogenesis. Our proteomics results not only confirmed our previous transcriptomic results on the role of the polyamine metabolic pathways and stress responses in cotton somatic embryogenesis, but identified key proteins important for cotton somatic embryogenesis and provided new insights into the molecular mechanisms underlying somatic embryogenesis in cotton.

Metastatic retinoblastoma of the parotid and submandibular glands: a rare case report.

BACKGROUND: Retinoblastoma is the most common intraocular malignancy occurring in children. It can metastasize to the regional lymph nodes, central nervous system and distant organs usually the bones and bone marrow and very rarely to the soft tissue. Here, we report a case of unilateral retinoblastoma in a 4-year-old girl accompanied by a large metastasis of the parotid and submandibular glands that developed about 6 months previously and gradually increased in size 5 months after enucleation of the left eye. CASE PRESENTATION: A 4-year-old girl with a history of unilateral retinoblastoma presented with a large, painful and worsening mass (about 20 × 23 cm) of the left side of the neck. Following surgery, the orbital tumour was completely resected, and the large tumour invasion range in the left side of the neck was not resected completely. Histopathological examination revealed retinoblastoma of the orbit and the parotid and submandibular glands. After chemotherapy and additional local radiotherapy on the parotid and submandibular glands, the tumour was inactive and stable. CONCLUSIONS: Delayed detection and inappropriate management contribute to poor outcomes. Fundus examinations, education regarding the early signs of RB, and optimization of the therapeutic strategy for RB may play important roles in ocular health.

Ras-related C3 botulinum toxin substrate 1 activation is involved in the pathogenesis of diabetic retinopathy.

This study used a streptozotocin (STZ)-induced rat model of diabetes to investigate whether Ras-related C3 botulinum toxin substrate 1 (Rac1) was involved in the pathogenesis of diabetic retinopathy. The effects of Rac1 inhibition on vascular endothelial (VE)-cadherin and ß-catenin expression in high glucose-induced rat retinal endothelial cells (RRECs) were additionally examined. Rac1 activation in the retinas from STZ-induced diabetic rats and in high glucose-induced RRECs was measured by reverse transcription-quantitative polymerase chain reaction analysis, immunohistochemistry and western blot analysis. The expression levels of VE-cadherin and ß-catenin were also examined with or without Rac1 inhibition through small interfering (si)RNA transfection. STZ-induced diabetes was associated with an increase in the vascular permeability of the retina. Furthermore, Rac1 activation was increased in the retina of STZ-induced diabetic rats and in high glucose-induced RRECs compared with that in the controls. Immunohistochemistry showed that immunostaining of Rac1 was localized in the outer plexiform, inner nuclear, inner plexiform and ganglion cell layers and in the retinal microvasculature of rats. The expression of ß-catenin was increased in the retinas of the diabetic rats at four, eight and 12 weeks after the induction of diabetes compared with that in the controls. Additionally, Rac1 activation was required for the high glucose-induced VE-cadherin expression decrease and for ß-catenin expression in high glucose-induced RRECs. Rac1 inhibition by Rac1-siRNA transfection effectively prevented hyperpermeability, ß-catenin expression and the VE-cadherin expression decrease in high glucose-induced RRECs. In conclusion, diabetes affects the expression of Rac1 in the retina. Rac1 may be involved in the diabetes-induced damage and/or alterations to the blood-retinal barrier through changes in VE-cadherin and ß-catenin expression.

[A new method of anti-jamming ability improvement for Michelson Interferometer].

In order to improve anti-jamming capability of Michelson interferometer system, replace the traditional structure of the moving mirror scanning was replaced, an interference system based on electro-optic modulation of crystal refractive index was designed to achieve optical path scanning. The system modulated voltage signal on the variable refractive crystal, to generate cyclical changes, changed the refractive index to control optical path difference in the original optical path system. Using electronic scanning to replace of mechanical scanning, improved the system's noise immunity was improved. In the electro-optic modulation process, computed the maximum optical path difference of the system was computed, and analyzed of the crystal thickness and crystal diffraction efficiency of the modulation process were analyzed. The simulation experiment shows that, with the modulation voltage range increasing, the available range of the optical path is also increased, and the system spectrum resolving power will also increase accordingly. Meanwhile, in the modulation process set the modulation range was set to make the energy of diffraction energy losses less than 10% of the total energy, so as to ensure a better signal to noise ratio. Experimental results show that, as the modulation voltage changes, interference fringes occurred continuously moved. When the voltage is further increased, the nonlinear error appears. After non-linear error correction for the system, spectrum resolution reached to 7. 2 cm-1, slightly lower than the original system. But its anti-jamming capability is greatly enhanced, as in the absence of experimental platform for seismic conditions, conventional interferometer relative error is more than 20%, while the relative error of the system is less than 5%, in line with the design requirements. It was proved that the anti-jamming capability of the system was enhanced greatly, when the static electro-optical modulation was used.

[Research on detecting concentration of serum protein based on resonance Rayleigh scattering].

The resonance Rayleigh scattering spectral detection system was designed based on the 2, 9, 16, 23-tetracarboxylate-phthalocyanine zinc and protein system. In the system, excitation light source is 405 nm wide band gap semiconductor lasers, and monochromator is 475 nm narrow-band band-pass filter, and the detector is low-noise and high-gain photoelectric amplifier based on blue-ray enhanced photodiode. Experiment shows that, the solution's strong absorption wavelength is near 420 nm. Under the action of incentive light, resonance Rayleigh scattering is generated at the resonant wavelength, and the scattering intensity is proportional to the protein content. The system uses 2, 9, 16, 23-tetracarboxylate as the spectrum probe to determine the concentration of serum proteins by resonance Rayleigh scattering method. Its linear detection range is 10 - 50 mg.mL-1, and its detection limit is 0. 001 mg.mL-1. The newly developed device for detecting concentration of the serum protein has the advantages of small size, low cost, low power consumption, and being easy to use.

[Research on formaldehyde concentration detection system based on photo-elastic modulation].

There is a growing number of environmental pollution caused by excessive indoor formaldehyde, and in order to quickly and accurately quantify the concentration of formaldehyde gas in indoor air, a system for detecting the concentration of formaldehyde gas based on photo-elastic modulation was designed. It consists of the infrared light source, filters, elastic light modulator, and infrared detectors, and photo-elastic crystal refractive index of cyclical changes was controlled by elastic light modulator. Refractive index caused by the changes in the optical path provided a spectrum distribution function of the optical path difference. Optical path difference function of the system was derived through the HITRAN spectral database. Experiments were carried out using infrared light source combined with narrow band filters, and the transmittance of the center wavelength was more than 90%. Photo-elastic crystal is ZnSe crystal as photo-elastic light modulator, and the drive frequency of the system is 100 kHz. For three different environments at different locations, 10 groups of sample gas were collected for analysis, and the concentration of formaldehyde gas was detected using standard spectrometer and the system for comparing the test data. Experimental results show that when the concentration of formaldehyde gas is high, the system performance is good; When the concentration of formaldehyde gas is low, the signal-to-noise ratio of the system is decreased, and the detection accuracy is slightly reduced, but it still meets the design requirements.

Primary malignant melanoma of the lacrimal sac.

Primary malignant melanoma of the lacrimal sac is extremely rare. It is usually diagnosed at an advanced stage after excision or biopsy of a tumour. We treated a 59-year-old man with tearing and bloody discharge from the right eye. Clinical examination revealed a firm, localised mass at the inner canthus consistent with a lacrimal sac swelling. Sac washout demonstrated obstruction to entry into the lacrimal sac with a reflux of blood-stained fluid. The pathological findings and the immunohistochemical studies showed a malignant melanoma of the lacrimal sac. We performed radical surgery and radiation therapy. Follow-up 4 months after surgery revealed no evidence of recurrence. Because this tumour often presents with symptoms similar to dacryocystitis and may masquerade as a chronic dacryocystitis, ophthalmologists should be aware of this disease entity when encountered with patients with epiphora and mass in the medial canthal area.

Application of the radial basis function interpolation to phase extraction from a single electronic speckle pattern interferometric fringe.

In this paper, we introduce the radial basis function (RBF) interpolation method to electronic speckle pattern interferometry (ESPI) and propose the RBF interpolation method to obtain unwrapped phase values based on a skeleton map. Because of the excellent approximation properties of the RBF interpolation, the proposed method can extract accurate phase values from a single fringe pattern effectively, even using a simple 3×3 mean filter as preprocessing. Using our method, both special filtering methods for ESPI fringes as preprocessing and postprocessing, including a dilatation and erosion algorithm for pruning and connecting and the smooth algorithm for improving the phase values are not needed. We test our method on a computer-simulated and two experimentally obtained poor-quality fringe patterns. The results have demonstrated that our RBF interpolation method works well even under a seriously disconnected skeleton map where it is impossible to apply the widely used, Matlab function grid data interpolation or the backpropagation neural networks method [Appl. Opt. 46, 7475 (2007)].

Integrin ß1 subunit signaling is involved in the directed migration of human retinal pigment epithelial cells following electric field stimulation.

AIMS: Direct current electric fields (EFs) can induce directed cell migration in a wide variety of cells, and this has been proven to be of importance in wound healing. Here we observed the effects of EFs on cultured human retinal pigment epithelial (RPE) cells and explored the possible involvement of integrin ß1 subunit signaling in the process. METHODS: Cultured human RPE cells were exposed to an EF at 5 V/cm for 3 h. The rate and directionality of cell migration were quantified. The distribution of integrin ß1 subunit was measured by immunohistochemistry and the expression of integrin ß1 subunit and phosphorylated focal adhesion kinase (FAK) was determined by PCR and Western blotting. Experiments were performed in the presence or absence of anti-integrin ß1 subunit antibody. RESULTS: During exposure to an EF at 5 V/cm for 3 h, the separated human RPE cells and wounded RPE monolayer demonstrated a cathodal-directed migration. The distribution of integrin ß1 subunit in the cells was also polarized to the cathode, and the expression in mRNA and its protein level were obviously increased. Furthermore, exposure to EFs of 5 V/cm triggered the phosphorylation of FAK in human RPE cells. In contrast, blocking of integrin ß1 subunit suppressed the directed migration of RPE cells and reduced the activation of FAK in EFs. CONCLUSIONS: These findings indicate that EF exposure results in directed migration of the separated RPE cells and RPE monolayer. These effects may partially act through the activation of integrin ß1 subunit signaling.

[Research on detecting trace formaldehyde gas by the multi-wavelengths characteristics method].

In order to overcome the slow speed of detecting trace formaldehyde in the sample gas, material consumption by chemical reaction, and the limitations of the sampling area in the detection of trace formaldehyde, a multi-wavelength characteristics method for getting the exact concentration of formaldehyde quickly was designed. According to the spectrum characteristics of formaldehyde and the main interfering gases the system chose multiple wavelengths with the minimum degree of coherence (the number of characteristic wavelengths were selected to be 3, 4 and 5), in conjunction with the corresponding groups of narrow-band filters. With the infrared light of the light source through the chamber windows and narrow-band filters, the infrared light was collected by the PCI-2TE-13 infrared detectors, and the concentration of formaldehyde in the sample gas was calculated by the characteristics spectrum absorption algorithm. In the experiments, the system analyzed and calculated the concentration of formaldehyde in four gas samples collected in the newly renovated house, building materials market, supermarkets and outdoor parks. Experimental results of the system and test results of ARCSpectro-AMIR infrared spectrometer were compared, the results show that test data above 10 mg x m(-3) were close to true value by the multi-wavelengths characteristics method, and the average error is less than 5%. So the system meets the requirements of practical applications, and it has the advantages of real-time detection, not poisoning so on.

Up-regulation of integrin-linked kinase in the streptozotocin-induced diabetic rat retina.

OBJECTIVES: This study investigated whether integrin-linked kinase (ILK) is involved in the pathogenesis of diabetic retinopathy, by analyzing the expression and activity of ILK in the retina from a streptozotocin (STZ)-induced rat model of diabetes. METHODS: ILK expression in the retina from both control and STZ-induced diabetic rats was measured by reverse transcription polymerase chain reaction, immunohistochemistry and Western blot analysis. The expressions of Akt and FOXO1A, the downstream molecules of ILK, were also examined. RESULTS: The present study showed that the STZ-induced diabetes was associated with the increase in the vascular permeability in the retina. This elevated vascular permeability increased with the progression of diabetic retinopathy. Meanwhile, the results also showed that the expression of ILK increased in protein and mRNA levels in the retina of STZ-induced diabetic rats. Immunohistochemistry showed that immunostaining of ILK was localized in the outer plexiform layer (OPL), the inner nuclear layer (INL), the inner plexiform layer (IPL), the ganglion cell layer (GCL) and the retinal microvasculature of rats. However, the expression of Akt was reduced in the retinas at 8 and 12 weeks and increased in the retinas at 4 weeks after induction of diabetes. Meanwhile, the expression of the FOXO1A protein increased in the retinas at 8 and 12 weeks and decreased in the retinas at 4 weeks after induction of diabetes. The FOXO1A immunostaining was also observed in the retinal microvasculature and in the OPL, INL, IPL and GCL of rat retinas. CONCLUSION: These results indicate that diabetes affects the expression of ILK in the retina. ILK may be involved in the diabetes-induced damage and/or alterations of neural and microvascular structures.

Epidermal growth factor receptor in cultured human retinal pigment epithelial cells.

BACKGROUND: Migration and proliferation of retinal pigment epithelial (RPE) cells play an important role in proliferative vitreoretinopathy. Epidermal growth factor receptor (EGFR) is a cell surface receptor with intrinsic tyrosine kinase activity. The engagement of the receptor by its ligand can induce intracellular mitogenic signal transduction pathways and stimulate proliferation, migration and differentiation of cells. This experiment aimed to investigate the activation and role of EGFR signal transduction pathway in proliferation of human RPE cells. METHODS: Cultured human RPE cells of the 3rd to 6th passages were studied by colorimetric assay for cellular growth and survival (MTT assay) to test the effects of EGF (0.1, 1, 10, 50, and 100 ng/ml) and fetal bovine serum (FBS) on proliferation of human RPE cells. An in vitro wound healing model was also set up, and the number of cells that had entered the denuded area was counted. The human RPE cells were cultured for 3 days with 0.1% FBS, 10% FBS, 10 ng/ml EGF + 0.1% FBS and a combination of EGF and 10% FBS, respectively. Immunohistochemical staining and in situ hybridization were used to observe the expressions of EGFR protein and mRNA, respectively. Activation of mitogen-activated protein kinase (MAPK) was detected by immunohistochemical method with specific antiphosphorylated extracellular signal-regulated kinase (ERK)1/2 antibody. RESULTS: EGF stimulated proliferation and migration of cultured human RPE cells in a concentration-dependent manner. The maximum of the proliferation rate of RPE cells was 81.8% with EGF at a concentration of 10-100 ng/ml of EGF in serum-free Dulbecco's modified essential medium (DMEM) and 122.7% at a concentration of 1-10 ng/ml of EGF in 5% FBS DMEM (p < 0.001); there was a significant difference between serum-free DMEM groups and 5% FBS DMEM groups. The maximum of the migration rate of the cells was 438.9% at a concentration of 10-100 ng/ml of EGF in 10% FBS DMEM, 147% with 10% FBS, and only 36% with EGF in 0.1% FBS at the concentration of 10 ng/ml (p < 0.001). EGF promoted the expression of EGFR protein and mRNA in RPE cells. FBS cooperated with EGF in the stimulation of EGFR expression, and it had a stronger effect in the process than EGF alone. After 3 days of incubation with EGF, phosphorylated ERK1/2 was detectable in the nucleus of RPE cells, whereas cells presented immunostaining positive for phosphorylated ERK1/2 in the cytoplasm before stimulation, indicating that EGF could induce MAPK nuclear translocation. CONCLUSION: EGF could induce EGF-EGFR-MAPK signal transduction pathway in human RPE cells in a concentration-dependent manner in vitro, which may play a key role in the activation of human RPE cell proliferation and migration.

[Effects of extracts of ginkgo biloba (EGB) on levels of nitric oxide and apoptosis in the retina induced by glutamate in adult rabbits].

OBJECTIVE: To study the effects of extracts of ginkgo biloba (EGB) on the levels of nitric oxide (NO) and apoptosis in the retina induced by glutamate by intravitreal injection in adult rabbit. METHOD: The model of apoptosis in retina induced by glutamate by intravitreal injection was established in adult rabbit and EGB was retrobulbarly injected. The levels of NO were measured by spectrophotometer. Retina DNA fragmentation was determined by agarose gel electrophoresis. RESULT: The levels of NO in retina in experiment groups were significantly increased compared with controls, after treatment with high dosage of EGB, and levels of NO was decreased to normal. They were not decreased by injection of small dosage of EGB. DNA fragmentation of retina apoptosis was detected in experiment groups and small dosage EGB groups. CONCLUSION: The retina apoptosis was induced by glutamate by intravitreal injection in adult rabbit and was inhibited by EGB that may be through blocking the generation of NO free radicals.