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.

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.

[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.

[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.