Reduced expression of the lncRNA NRON is a potential hallmark of the CMV-amplified CD8+ T cell accumulations commonly seen in older humans.

The characteristic accumulation of late-stage differentiated CD8+ T cells is enhanced by lifelong latent cytomegalovirus (CMV) persistence, which makes it challenging to screen for subclinical biomarkers of immune aging in the elderly. We systematically identified predominantly preformed, long, noncoding RNAs (lncRNAs) as integrative biomarkers of CD8+ T cell aging in 14 elderly CMV carriers over 80 years of age. After sorting the CD28nullCD8+ T cell subset and its CD28bearingCD8+ counterpart in five nonagenarians, we profiled the differential expression of lncRNAs and genes in CD28nullCD8+ T cells via array detection. We focused on 11 differentially expressed antisense lncRNAs and cross-referenced them with previously identified age-accumulated lncRNAs to create a set of candidates in CD28nullCD8+T cells. We performed intracellular validation on the age-accumulated candidate lncRNAs paired with their antisense target genes using quantitative polymerase chain reaction (qPCR). Simultaneously, we sorted the CMVpp65-specific CD8+ T cell subset and its counterpart from participant cells with the HLA-A-*0201 genotype. The validated age-accumulated lncRNAs in CD28nullCD8+ T cells were intracellularly cross-validated in CMVpp65CD8+ T cells. Finally, we identified the immunity-related gene(s) that acted as potential target(s) to the cross-validated age-accumulated lncRNA(s), using bioinformatics techniques. The potential regulation of the final identified lncRNA-gene pair(s) was simultaneously predicted in two pathway-integrated networks. We concluded that expression of an age-accumulated lncRNA (NRON) was decreased, whereas that of its immunity-related target gene (NFAT) was increased, in both CD28nullCD8+ T cells and CMVpp65CD8+ T cells of elderly individuals with persistent CMV infection. The identification of NRON as a potential biomarker suggests that NRON contributes to CMV-enhanced CD28nullCD8+ T cell aging by modulating phosphorylation and/or IL-4-dependent NFAT signaling.

Lack of association of C3 gene with uveitis: additional insights into the genetic profile of uveitis regarding complement pathway genes.

Uveitis is a devastating ocular disease that causes blindness. Our previous studies have achieved great advancements in depicting the genetic profiles of uveitis regarding complement pathway genes. This study aimed to provide additional insights into this interest by testing the "central" factor of the complement system, C3 gene variants, in two uveitis entities. Eight haplotype-tagging SNPs of C3 gene were genotyped in 141 anterior uveitis (AU), 158 non-infectious intermediate and posterior uveitis (NIPU) and 293 controls. The results showed that none of the tagging SNPs had a significant association with uveitis (P > 0.05), either in the global uveitis or subtypes. Although rs428453 showed a nominal association with NIPU subtype in the recessive model (P = 0.042), the P value could not withstand the Bonferroni correction (P corr > 0.05). Stratification analyses according to HLA-B27 status and correlation analysis still did not find any significant interactions or genetic markers regarding AU. Logistic regression analysis also revealed no gender-related epistatic effects of C3 on uveitis. Two haplotype blocks were defined across the C3 locus but neither of them was significantly associated with uveitis or subtypes. This study shows no significant association of the C3 gene with uveitis, suggesting C3 confers either no or limited risk for uveitis susceptibility.

Comprehensive circular RNA profiling reveals that circular RNA100783 is involved in chronic CD28-associated CD8(+)T cell ageing.

BACKGROUND: Ageing brings about the gradual deterioration of the immune system, also known as immunosenescence. The role of non-coding circular RNA in immunosenescence is under studied. Using circular RNA microarray data, we assembled Comparison groups (C1, C2, C3 and C4) that allowed us to compare the circular RNA expression profiles between CD28(+)CD8(+) T cells and CD28(-)CD8(+) T cells isolated from healthy elderly or adult control subjects. Using a step-wise biomathematical strategy, the differentially-expressed circRNAs were identified in C1 (CD28(+)CD8(+) vs CD28(-)CD8(+)T cells in the elderly) and C4 (CD28(-)CD8(+)T cells in the elderly vs in the adult), and the commonly-expressed circRNA species from these profiles were optimized as immunosenescence biomarkers. RESULTS: Four overlapping upregulated circular RNAs (100550, 100783, 101328 and 102592) expressed in cross-comparison between C1 and C4 were validated using quantitative polymerase chain reaction. Of these, only circular RNA100783 exhibited significant validation. None of the down-regulated circular RNAs were expressed in the C1 and the C4 cross-comparisons. Therefore, we further predicted circular RNA100783-targeted miRNA-gene interactions using online DAVID annotation. The analysis revealed that a circular RNA100783-targeted miRNA-mRNA network may be involved in alternative splicing, the production of splice variants, and in the regulation of phosphoprotein expression. Considering the hypothesis of splicing-related biogenesis of circRNAs, we propose that circular RNA100783 may play a role in phosphoprotein-associated functions duringCD28-related CD8(+) T cell ageing. CONCLUSIONS: This study is the first to employ circular RNA profiling to investigate circular RNA-micro RNA interactions in ageing human CD8(+)T cell populations and the accompanying loss of CD28 expression. The overlapping expression of circular RNA100783 may represent a novel biomarker for the longitudinal tracking ofCD28-related CD8(+) T cell ageing and global immunosenescence.

[Clinical characteristics and pathogenesis of intracapillary hemorrhage with adjacent peripapillary subretinal hemorrhage].

OBJECTIVE: To study the clinical characteristics and pathogenesis of intracapillary hemorrhage with adjacent peripapillary subretinal hemorrhage (IHAPSH). METHODS: A retrospective review of 8 patients with IHAPSH. The demographical data of the patients, the symptoms, initial and final visual acuities, biomicroscopic findings, fundus photographs, and the results of various special examinations, including fluoresce in angiography, B-scan ultrasonography, optical coherence tomography (OCT) and CT were analyzed. RESULTS: Fundus examination of 8 patients showed various changes of optic disc, including: a small and tilted optic disc with elevation and blurring of the nasal or superonasal edge. Hemorrhage of the optic disc was present in the cup and/or on the nasal edge, extended to the surface of the optic disc, the superficial layer of the retina and the subretinal space. Subretinal hemorrhage was located on the nasal, superior and inferior side of the optic disc. OCT examination showed an elevated optic nerve head with an enlarged nasal peripapillary subretinal space. CONCLUSIONS: In the IHAPSH patient, bleeding may originate from the posterior ciliary artery that traverse the sclera window of the disc, passes through the space between the optic nerve and the scleral canal, reaches the edge of the optic disc and the subretinal space, penetrates into the optic nerve tissues anterior to the lamina cribrosa, reaches the optic cup and extends to the surface of the optic disc, the retina or the vitreous.

[Prevalence of diabetic retinopathy among the elderly in rural southern Shuangcheng city, Heilongjiang province].

OBJECTIVE: The purpose of this study was to understand the awareness, prevalence of diabetic retinopathy and treatment status of people aged over 50 and living in the rural areas of Shuangcheng city, Heilongjiang province, China. METHODS: Cluster sampling was used in randomly selected 5504 survey for ophthalmic clinical examination, in patients with diabetic retinopathy. A questionnaire in the state of knowledge about prevention and treatment was developed. RESULTS: Among the 5504 persons entering in the project, 5053 were examined on their eyes (91.8%). In this selected population, 56 persons (112 eyes) were diagnosed as diabetic retinopathy (1.108%), with 95% confidence interval (CI) as: 0.819% to 1.397%. Of 56 patients, 49 cases were non-proliferative diabetic retinopathy, accounting for 87.50% of the total number of patients with diabetic retinopathy; proliferative diabetic retinopathy 7 cases, accounting for 12.50% of the 112 eyes, 6.25% (7/112) having vitreous hemorrhage, 8.04% (9/112) having macular edema. For diabetic retinopathy prevalence rates, there was no significant difference in males and females. Between the per differential 10-year-old division, the difference was significant. Among the 60 to 69 group, a significantly higher prevalence rate was seen. Of the 112 eyes with diabetic retinopathy, 34 eyes (30.4%) were low vision [visual acuity < 20/60 (0.3) to ≥ 20/400 (0.05)]; 6 eyes (5.4%) were blind [visual acuity < 20/400 (0.05) to NLP]. The rate in the patients with PDR and fasting blood glucose was above 11.1 mmol/L was higher than having NPDR and fasting blood glucose below 11.1 mmol/L. Having fasting blood glucose 11.1 mmol/L and above with the course over five years among patients with PDR, the proportion of fasting blood glucose was higher than those with less than 11.1 mmol/L and diabetic retinopathy duration of less than five years. Of 56 patients with diabetic retinopathy, 38 cases (67.9%) did not receive any treatment. Among 18 cases (32.1%) with insulin or oral drug therapy, regularly using insulin or other medication (14.3%), only 1 (1.8%) case was given the treatment for diabetic retinopathy. Results from our survey showed that patients with diabetic retinopathy had a poor understanding about prevention and treatment of the disease. CONCLUSION: Long duration and high blood glucose in patients with diabetic retinopathy seemed to be the important risk factor. Early systematic drug use for prevention and blood glucose control was the key to prevent diabetic retinopathy. Patients with diabetic retinopathy in China had poor understanding about the prevention measures of the disease and lack of knowledge.

[Prevention of posterior capsule opacification by combination of arsenic trioxide and perfect capsule].

OBJECTIVE: Despite recent improvements in IOL design, posterior capsule opacification (PCO) remains a significant clinical problem after cataract surgery. The Perfect Capsule device (Milvella, Ltd., Epping, Australia) permits the introduction and subsequent removal of potentially toxic agents into the closed capsular bag. The present purpose was to detect the effectiveness of arsenic trioxide (As2O3) to cultured human lens cells and cells within the human capsular bag. METHODS: All experiments were carried out with 2 minutes exposure to As2O3. Effect of As2O3 on FHL124 cells growth was tested by MTT. Changes in cell calcium levels induced by high concentration of As2O3 were measured by real-time fluorometric single-cell digital imaging techniques after Fura-2 incorporation. In vitro human capsular bags were also tested after a sham surgery was performed using the Perfect Capsule device to form a closed system. As2O3 (10, 30, 100, 300, 1000 µmol/L) were used with donor match-pairs treated with medium alone.(n = 3 in all cases).On-going observations were by phase-contrast microscopy. Cellular architecture was examined by fluorescence cytochemistry. RESULTS: As2O3 of 100 µmol/L and above inhibited the growth of FHL 124 cells in a dose-dependent manner with 2 minutes exposure (t = 5.217, P < 0.01, IC(50) is 130 µmol/L) As2O3 depleted the calcium store and consequently lead to a loss of calcium signaling.Moreover, 1 × 10(4) µmol/L As2O3 had a moderate effect on the calcium influx pathway, inhibition on the amplitude and rapidity is (43.24 ± 2.98)% (t = 3.134, P < 0.01) and (46.27 ± 6.01)% (t = 3.521, P < 0.01), respectively. No cell left in the vitro human capsular bags after being treated with 1 × 10(4) µmol/L As2O3. CONCLUSIONS: As2O3 can eliminate the human lens epithelium cells remained in the capsular bag in cataract surgery, which may help to prevent PCO. The application of As2O3 to cells within the capsular bag for a 2 minute window using the Perfect Capsule system predicts this compound will have therapeutic benefit to humans in vivo.

Time-dependent reduction of glutamine synthetase in retina of diabetic rats.

Diabetic Retinopathy (DR) is one of the most common complications of diabetes and a major cause of blindness worldwide. We studied the transcriptome of the diabetic retina using Series Analysis of Gene Expression (SAGE) technology and observed a 45.6% reduction in transcript levels of glutamine synthetase (GS) in streptozotocin-induced diabetic rats compared with normal rats. RT-PCR and colorimetric enzyme activity assays revealed significant differences in GS mRNA expression and enzyme activity as early as the first month of diabetes development, with a progressive decrease in GS mRNA level and enzyme activity over a 12-month period. Northern blot analysis indicated a linear correlation between the reduction in GS expression and the time course of diabetic retinopathy (r = 0.802, p < 0.0001), which was validated by real-time RT-PCR (r = 0.731, p < 0.001). Our results implicate GS as a possible biomarker for evaluating the severity of developed diabetic retinopathy over the time course of diabetes progression.

[Mechanism of arsenic trioxide induced apoptosis in cultured human lens epithelium cells].

OBJECTIVE: To study cytotoxic effects of arsenic trioxide (As2O3) on the human lens epithelium cells and to identify the biological mechanism for these effects. METHODS: In this experimental study, human lens epithelium cells (FHL124 cells) were cultured in Eagle's minimum essential medium supplemented with 5% fetal calf serum. The effects of As2O3 on FHL124 cells growth were tested by MTT, and apoptosis was detected by TUNEL assay. Gene changes were detected by real-time PCR (Taqman). As2O3-induced changes in cell calcium level were measured by real-time fluorometric single-cell digital imaging techniques after Fura-2 incorporation. RESULTS: As2O3 inhibited the growth of FHL 124 cells in vitro in a dose-dependent manner, given an IC50 value of 1.5 micromol/L. As2O3 induced apoptosis of FHL124 cells as showed by TUNEL assay. As2O3 provoked an endoplasmic reticulum (ER) stress response identified through an up regulation of EIF2A, ERN1 and ATF6 (F = 8.51, P = 0.0005). As2O3 depleted the calcium store and consequently lead to a decrease of calcium signaling (P = 0.0018). Moreover, As2O3 had a moderate effect on the calcium influx pathway. CONCLUSIONS: As2O3 inhibits the growth and induces apoptosis of human lens epithelium cells. As2O3 provokes an ER stress which could be the cause of apoptic processes.