Exome sequencing and genome-wide association analyses unveils the genetic predisposition in hydroxychloroquine retinopathy.

OBJECTIVES: To unveil the candidate susceptibility genes in chloroquine/hydroxychloroquine (CQ/HCQ) retinopathy using whole exome sequencing (WES) and genome-wide association study (GWAS). METHODS: Patients with a diagnosis of CQ/HCQ retinopathy based on the comprehensive demographic and ocular examination were included. The peripheral blood was extracted for WES and GWAS analyses. The Chinese Han Southern database from 1000 genomes was used as control group to compare the affected percentage. Multivariate logistic regression analysis adjusted for age, HCQ dose, duration and renal disease were used to analyze the correlation between genetic variants and visual outcome. A poor vision outcome was defined as visual acuity <6/12. An abnormal anatomical outcome was defined as disruption of ellipsoid zone in the fovea. RESULTS: Twenty-nine patients with an average age of 60.9 ± 13.4 years, treatment duration of 12.1 ± 6.2 years, daily dose of 8.5 ± 4.1 mg/kg, and the cumulative dose of 1637.5 ± 772.5 g, were genotyped. Several candidate genes associated with CQ/HCQ retinopathy were found, including RP1L1, RPGR and RPE65, with a difference of affected percentage over 50% in mutation between the case and control groups. New foci in CCDC66: rs56616026 (OR = 63.43, p = 1.63 × 10-8) and rs56616023 (OR = 104.7, p = 5.02 × 10-10) were identified significantly associated with HCQ retinopathy. Multivariate analysis revealed increased genetic variants were significantly associated with poor functional (OR = 1.600, p = 0.004) and structural outcome (OR = 1.318, p = 0.043). CONCLUSIONS: Several candidate susceptibility genes including RP1L1, RPGR, RPE65 and CCDC66 were identified to be associated with CQ/HCQ retinopathy. In addition to disease susceptibility, patients with increased genetic variants are more vulnerable to poor visual outcomes.

Microsporidial keratitis in patients with hot springs exposure.

This retrospective study included 10 eyes of 9 patients diagnosed with microsporidial keratitis. All of them were known to contract this disease after taking baths in hot springs. The disease was diagnosed based on detecting microsporidia in corneal scrapings using Gram stain and the modified Kinyoun's acid-fast stain. The specimens from the last six patients were subjected to PCR and then sequencing. All of them revealed that the microorganism identified has a high similarity to Vittaforma corneae. Repeated debridement of the epithelial lesions successfully eradicated the microsporidial infection in all nine patients.

Desipramine activated Bcl-2 expression and inhibited lipopolysaccharide-induced apoptosis in hippocampus-derived adult neural stem cells.

Desipramine (DP) is a tricyclic antidepressant used for treating depression and numerous other psychiatric disorders. Recent studies have shown that DP can promote neurogenesis and improve the survival rate of hippocampal neurons. However, whether DP induces neuroprotection or promotes the differentiation of neural stem cells (NSCs) needs to be elucidated. In this study, we cultured NSCs derived from the hippocampal tissues of adult rats as an in vitro model to evaluate the modulation effect of DP on NSCs. First, we demonstrated that the expression of Bcl-2 mRNA and nestin in 2 microM DP-treated NSCs were up-regulated and detected by real-time reverse transcriptase polymerase chain reaction (RT-PCR). The results of Western blotting and immunofluorescent study confirmed that Bcl-2 protein expression was significantly increased in Day 3 DP-treated NSCs. Using the Bcl-2 small interfering RNA (siRNA) method, our results further showed that DP protects the lipopolysaccharide (LPS)-induced apoptosis in NSCs, in part by activating the expression of Bcl-2. Furthermore, DP treatment significantly inhibited the induction of proinflammatory factor interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha in the culture medium of LPS-treated NSCs mediated by Bcl-2 modulation. The results of high performance liquid chromatography coupled to electrochemical detection further confirmed that DP significantly increased the functional production of serotonin (26+/-3.5 microM, DP-treated 96 h) and noradrenaline (50+/-8.9 microM, DP-treated 96 h) in NSCs through activation of the MAPK/ERK pathway and partially mediated by Bcl-2. In conclusion, the present results indicate that DP can increase neuroprotection ability by inhibiting the LPS-induced inflammatory process in NSCs via the modulation of Bcl-2 expression, as confirmed by the siRNA method.

Detection of HIV RNA levels in intraocular and cerebrospinal fluids in patients with AIDS-related cryptococcosis.

The objective of the present study was to evaluate human immunodeficiency virus (HIV) levels in the aqueous humor and vitreous fluid in patients with and those without ocular involvement due to AIDS-related cryptococcosis. We also assessed whether cryptococcosis infection in the central nervous system (CNS) was associated with elevated HIV levels in the cerebrospinal fluid (CSF). From 1993 to 2003, we obtained 16 CSF samples from 9 AIDS patients with cryptococcal meningitis and 7 AIDS patients without CNS opportunistic infection. Samples of intraocular fluids were obtained from all 9 patients with cryptococcal meningitis. Five cases presented with ocular involvement in patients with meningitis. By using the method of reverse transcriptase-polymerase chain reaction, we detected higher HIV loads in aqueous humor (27,244+/-4,123 copies/ml) and vitreous fluid (84,930+/-5,071 copies/ml) in patients with concomitant CNS and ocular involvement due to AIDS-related cryptococcosis (p<0.05). HIV levels in the vitreous fluid were correlated with levels in CSF (r=0.77). Mean HIV level in the CSF (209,761+/-18,787 copies/ml) was significantly elevated in AIDS patients with cryptococcoal meningitis (p<0.05). To our knowledge, this is the first report to study the level of HIV loads in the CSF and intraocular fluid simultaneously in AIDS patients with cryptococcosis. Our results revealed the intrathecal and intraocular HIV replication in patients with cryptococcosis.

The HIV RNA Levels of Plasma and Ocular Fluids in AIDS Patients with Ophthalmic Infections.

The aim of the present study was to evaluate the impact of highly active antiretroviral therapy (HAART) on HIV viral load of plasma and intraocular fluids in AIDS patients with ophthalmic opportunistic infections. We further compared the treatment effect of HAART on these patients. From June 1997 to July 2003, we examined and followed up the ophthalmic conditions of 49 patients receiving HAART with ophthalmic diseases during this period. The method of reverse-transcriptase polymerase chain reaction was used to detect and monitor HIV load in plasma and/or aqueous humor of AIDS patients. Before HAART, the HIV levels in the plasma and aqueous humor in 8 AIDS patients with ophthalmic opportunistic infections were significantly higher than those in 6 patients with HIV-related retinopathy (p < 0.05). Compared to the eye findings and clinical improvement, HIV loads of aqueous humor in 10 of 14 AIDS patients (6 with HIV-related retinopathy, 5 with cytomegalovirus retinitis, 2 with toxoplasmic retinitis and 1 with cryptococcal chorioretinitis) declined to undetectable levels (< 400 copies/ml) after 4-8 months of HAART. HIV virus levels in the plasma of AIDS patients were significantly decreased, and the CD4 counts of these patients were significantly increased (Wilcoxon test) after initiation of HAART.

Pseudomonas corneal ulcer related to overnight orthokeratology.

PURPOSE: To report two cases of Pseudomonas aeruginosa corneal ulcers as a complication of overnight orthokeratology lens wear. METHODS: Case report. RESULTS: Two 11-year-old girls with acute central corneal ulcers were referred to our hospital. In both cases, the ulcers were about 2 mm in diameter, located centrally, contained dense cellular infiltration, and discharged purulent material. Intensive topical ceftazidime was applied to treat the ulcers. Cultures of the scraped corneal tissues and the contact lens storage solutions in both cases grew P. aeruginosa, which was sensitive to the antibiotic. The presenting best-corrected visual acuity was hand motion at 20 cm in one patient and 6/20 in the other. Both patients had received several months of overnight orthokeratology treatment with rigid gas permeable contact lenses to correct myopia (-4.25 D and -4.75 D in the two affected eyes). The final best-corrected visual acuity was 6/60 in one patient and 6/7.5 in the other. CONCLUSIONS: Overnight orthokeratology contact lens wear carries a potential risk of corneal ulcer and may cause significant visual impairment in children.

Increased interleukin-6 in aqueous humor of a patient with lung-metastatic intraocular adenocarcinoma.

PURPOSE: We report increased interleukin (IL)-6 level in aqueous humor of a case of lung adenocarcinoma with metastasis to iris and anterior chamber. DESIGN: Interventional case report. METHODS: In one case and 11 healthy cataract patients as controls, we checked the levels of IL-6, IL-2, and tumor necrosis factor (TNF)-alpha in aqueous humor and serum by enzyme-linked immunosorbent assay. RESULTS: Levels of these three cytokines in serum were almost the same as with the levels in aqueous humor in the control group. The IL-6 concentrations ranged from 12.6 to 27.0 pg/ml (mean, 23.5 pg/ml) in serum and 15.0 to 43.1 pg/ml (mean, 22.4 pg/ml) in aqueous humor. There was no statistically significant difference between them. Compared with the control, the IL-6 concentration in aqueous humor of this patient, 282.0 pg/ml, increased more than 10-fold. CONCLUSION: This is a report describing elevated IL-6 level in the aqueous humor of a patient with intraocular metastasis from lung adenocarcinoma.

Nitric oxide levels in the aqueous humor in cataract patients.

PURPOSE: To determine the role of nitric oxide (NO) in the clinical manifestations of cataract in different age and etiological groups. SETTING: Multicenter study, Taipei, Taiwan, Republic of China. METHODS: Aqueous humor samples were obtained from 114 cataract patients in different etiological and age groups (eg, juvenile, traumatic, and age-related cataract). Using chemiluminescence assay, NO was detected through the light generated by ozone and NO interaction. The amount of NO production was calculated and compared among groups. The results were correlated with patient age. RESULTS: The NO levels in aqueous humor were highest in the 12 patients with traumatic cataract and lowest in the 15 patients with juvenile cataract (47.59 micromol/L +/- 12.81 [SD] and 7.66 +/- 2.62 micromol/L, respectively; P <.001). Aqueous humor NO levels were correlated with age and were highest in patients older than 80 years (mean 38.78 +/- 6.29 micromol/L) and in those with mature cataract (mean 40.15 +/- 6.15 micromol/L) (P <.05). CONCLUSIONS: The NO levels in the aqueous humor increased with age and traumatic events. The results suggest that NO could be a risk factor in cataract formation.