Xianchai Lin

Inhibition of Pirfenidone on TGF-beta2 Induced Proliferation, Migration and Epithlial-Mesenchymal Transition of Human Lens Epithelial Cells Line SRA01/04

Background Posterior capsular opacification (PCO) is a common complication of cataract surgery. Transforming growth factor-β2 (TGF-β2) plays important roles in the development of PCO. The existing pharmacological treatments are not satisfactory and can have toxic side effects. Methodologies/Principal Findings We evaluated the effect of pirfenidone on proliferation, migration and epithlial-mesenchymal transition of human lens epithelial cell line SRA01/04 (HLECs) in vitro. After treatment with 0, 0.25, and 0.5 mg/ml pirfenidone, cell proliferation was measured by MTT assay. Cell viability was determined by trypan blue exclusion assay and measurement of lactate dehydrogenase (LDH) activity released from the damaged cells. And cell migration was measured by scratch assay in the absence or presence of transforming growth factor-β2 (TGF-β2). The expressions of TGF-β2 and SMADs were evaluated with real-time RT-PCR, western blot, and immunofluorescence analyses. The mesenchymal phenotypic marker fibronectin (FN) was demonstrated by Immunocytofluorescence analyses. The cells had high viability, which did not vary across different concentrations of pirfenidone (0 [control] 0.3, 0.5 or 1.0 mg/ml) after 24 hours. Pirfenidone (0∼0.5 mg/ml) had no significant cytotoxicity effect on SRA01/04 by LDH assay. Pirfenidone significantly inhibited the proliferation and TGF-β2-induced cell migration and the effects were dose-dependent, and inhibited TGF-β2-induced fibroblastic phenotypes and TGF-β2-induced expression of FN in SRA01/04 cells. The cells showed dose-dependent decreases in mRNA and protein levels of TGF-β2 and SMADs. Pirfenidone also depressed the TGF-β2-induced expression of SMADs and blocked the nuclear translocation of SMADs in cells. Conclusion Pirfenidone inhibits TGF-β2-induced proliferation, migration and epithlial-mesenchymal transition of human lens epithelial cells line SRA01/04 at nontoxic concentrations. This effect may be achieved by down regulation of TGF-β/SAMD signaling in SRA01/04 cells.

Health Literacy, Computer Skills and Quality of Patient-Physician Communication in Chinese Patients with Cataract

Purpose The aim of the study was to assess levels of health literacy and computer skills in Chinese patients with cataract, and their impact on the doctor-patient relationship. Methods We undertook a cross-sectional study of cataract patients scheduled for cataract extraction procedures in Guangdong Province, China. Generic health literacy was assessed using 3 established screening questions. Adequate computer skills was determined if patients had used a computer and routinely used search engines on the Internet. Socio-demographic measures (e.g., age, sex, education) were obtained from a standardized interview. Participants who indicated that they could not understand what their doctors mean were considered to have had poor patient-physician communications. Results Of the 211 participants, 92 (43.6%) had inadequate health literacy and 204 (96.7%) inadequate computer skills. In multivariate analysis, females were more likely to have inadequate health literacy (odds ratio = 2.5, 95% confidence intervals [CI]: 1.3 to 4.7). People with inadequately health literacy were more likely to have a poor patient-physician communication (odds ratio = 3.5, 95% CIs: 1.3 to 9.0). Similar associations were found for inadequate computer skills. Conclusion Chinese elderly patients with cataract have inadequate health literacy and very limited computer skills, which place them at high risk of misunderstanding and mismanaging their ocular conditions. Patient education information other than online materials may improve the eye care and outcomes of these patients.

Dexamethasone Increases Cdc42 Expression in Human TM-1 Cells

Abstract Purpose: Changes in the cytoskeletal organization of the human trabecular meshwork (HTM) is thought to be responsible for primary open-angle glaucoma (POAG) pathologies. Cdc42 is a Rho GTPase; Rho GTPases are important modulatory agents of the cytoskeleton. This study aimed to investigate the effects of dexamethasone (DEX) on Cdc42 in a transformed HTM cell line, TM-1 to understand the molecular pathologies underlying POAG. Methods: TM-1 cells were cultured in vitro. The cultures were treated with DEX at 10−6 and 10−7 M for 1–4 days. Cdc42 was silenced using small interfering RNA (siRNA). The expression levels of Cdc42 in the TM-1 cells were measured using reverse transcription (RT)–PCR, western blotting analysis and immunofluorescence. Its downstream effectors, p21-activated kinase phosphorylation (phospho-PAK) and myosin light chain kinase (MLCK), were measured using western blotting analysis. In addition, the F-actin of TM-1 cells was stained using phalloidin. Results: The mRNA and protein levels of Cdc42 showed an increase in TM-1 cells with DEX treatment and a decrease in TM-1 cells transfected with Cdc42 siRNA. Moreover, phospho-PAK levels increased, whereas MLCK levels appeared to decrease, with DEX treatment. The F-actin of DEX-treated TM-1 cells displayed a rearrangement. Cdc42 siRNA decreased the expression of Cdc42 and its related proteins, resulting in an attenuation of the effects of DEX on Cdc42 and F-actin organization in TM-1 cells. Conclusions: DEX increases Cdc42 expression in TM-1. This may represent a potential mechanism of DEX-induced HTM cytoskeletal rearrangement.

Preoperative Expectations and Postoperative Outcomes of Visual Functioning among Cataract Patients in Urban Southern China.

Purpose To investigate the relationship between preoperative expectations and actual postoperative outcomes of visual function (VF) among patients undergoing first eye cataract surgery. Methods A longitudinal study of 182 patients from hospitals in urban Southern China were surveyed prior to surgery and 3 month after cataract surgery regarding their preoperative, expected postoperative and actual postoperative VF for each of the items on the Catquest-9SF and their satisfaction with cataract surgery. In addition, detailed clinical data were collected preoperatively and postoperatively. Results The majority of cataract patients in urban Southern China had high expectations for VF outcomes after cataract surgery and in most cases postoperative outcomes achieved the expected level of improvement. The mean (standard deviation, SD) preoperative Catquest-9SF score was 15.7 (5.86) and the mean (SD) expected postoperative score was 26.3 (2.93). The discrepancy between actual and expected improvement was significantly correlated with patients’ health literacy, presence of systemic and ocular comorbidity, preoperative visual acuity of the surgery eye, LOCS III nuclear opalescence and cortical cataract grading. Conclusion Cataract patients in urban Southern China had high expectations for surgery outcomes. Patients with low level of health literacy and the presence of systemic and ocular comorbidity may need a comprehensive counseling to decrease the discrepancy regarding expected and actual outcomes.

Expression of 14-3-3 Zeta Protein in Dexamethasone-Treated Mice and Human TM-1 Cells

ABSTRACT Purpose: 14-3-3 zeta protein plays a potential protective role in neurodegenerative disease. Given that glaucoma and neurodegenerative diseases share a similar pathogenesis, it is possible that 14-3-3 zeta may have a similar protective effect in the glaucomatous process. In the present study, we measured the expression of 14-3-3 zeta in vivo (mouse eyes) and in vitro in a transformed human trabecular meshwork (HTM) cell line, TM-1, and assessed the possible roles of this protein in dexamethasone (DEX)-treated eyes and HTM cells. Methods: Mouse eyes were randomly treated with 0.1% dexamethasone (DEX) eye drops or phosphate-buffered solution (PBS) for 28 days. The expression and distribution of 14-3-3 zeta protein in mouse eyes were examined using immunofluorescence. TM-1 cells were treated with DEX (10−6 or 10−7 M) or PBS for 1, 4, or 7 days, and the mRNA and protein expression of 14-3-3 zeta were detected by real-time RT-PCR and Western blotting. Results: 14-3-3 zeta protein was highly expressed in the mouse cornea, trabecular meshwork (TM), and ciliary body. Intraocular pressure (IOP) was significantly elevated, whereas the 14-3-3 zeta expression was significantly decreased in mouse TM after 0.1% DEX treatment for 28 days. In vitro, treatment with 10−7 M DEX mildly increased 14-3-3 zeta mRNA and protein expression (p > 0.05), whereas 10−6 M DEX significantly decreased expression of 14-3-3 zeta mRNA and protein (p < 0.05) compared to the control (Ctrl) group at the seventh day. Conclusions: DEX can increase IOP in mouse eyes and concurrently downregulate 14-3-3 zeta protein expression in mouse TM. The effects of DEX on 14-3-3 zeta expression in vitro were both dose- and time-related. Our results suggest that alterations in 14-3-3 zeta protein may be implicated in DEX-induced pathological elevated IOP.