Blockade of insulin receptor substrate-1 inhibits biological behavior of choroidal endothelial cells.

AIM: To investigate the effects of blockade of insulin receptor substrate-1 (IRS-1) on the bio-function of tube formation of human choroidal endothelial cells (HCECs). METHODS: Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to determine the expression level of IRS-1 and phospho-IRS-1 in HCECs. Tube formation of HCECs was analyzed using three dimensional in vitro Matrigel assay with or without IRS-1 blockage via IRS-1 inhibitor (GS-101) and vascular endothelial growth factor receptor 2 (VEGFR2) inhibitor. In addition, cell counting kit (CCK)-8 and Transwell migration assay were exerted to analyze the effects of blockade of IRS-1 on the bio-function of proliferation and migration of HCECs, respectively. The apoptosis of HCECs was examined using flow cytometry (FCM). RESULTS: RT-PCR and Western blot revealed that IRS-1 phospho-IRS-1 were expressed in HCECs and the expression level was enhanced by stimulation of VEGF-A. The number of tube formation was decreased significantly in GS-101 treated groups compared to phosphate buffered saline (PBS) treated control groups. Furthermore, both cell proliferation and migration of HCECs were decreased in the presence of GS-101. FCM analysis showed that the apoptosis of HCECs was enhanced when the cells were treated with GS-101. Western blot also showed that the expression level of cleaved-caspase 3 in GS-101 treated group was higher than that in control group. CONCLUSION: Blockade of IRS-1 can inhibit tube formation of HCECs through reducing cell proliferation and migration and promoting cell apoptosis.

[Eukaryotic expression and biological activity of recombinant human IL-17B protein].

AIM: To construct pCEP4/hIL-17B recombinant expression vector and express it stably in eukaryotic cells and investigate the biological activity in vitro. METHODS: The CDS region of human IL-17B gene was cloned by RT-PCR. After identification by sequencing, the hIL-17B gene was inserted into expression vector of pCEP4 to construct the recombinant vector pCEP4/hIL-17B, then transfected into 293T cells. The transgenic 293T cell line stably expressing rhIL-17B protein was selected in the presence of Hygromycin B. After FCS-free cultivation and sub-cloning, The IL-17B/mFc gene and protein expression was confirmed by RT-PCR, ELISA and Western blot analysis. To investigate the ability of combination with IL-17B receptor on human leukemic monocytic cell line, THP-1, by Flow cytometrical analysis (FACS) and of stimulation to secret cytokines in vitro. RESULTS: The recombinant pCEP4/hIL-17B and its transgenic 293T cells stably expressing rhIL-17B protein were obtained successfully. FACS analysis showed its high affinity with its receptor and it can stimulated THP-1 cell line to excrete IL-1ß and TNF-α in vitro and consistently caused a dose-dependent influx of neutrophil into the peritoneal cavity by intraperitoneal injection in vivo. CONCLUSION: The obtainment of transgenic 293T cell line stably expressing rhIL-17B protein paved the way for further study on biological functions of hIL-17B.

[Changes in monochromatic higher-order aberrations in different pupil sizes with accommodation in young emmetropes].

OBJECTIVE: To study the effect of accommodation on monochromatic higher-order aberrations in different pupil sizes in the young emmetropes. METHODS: Intervention trial design was used in the study. The monochromatic aberrations were measured in 12 eyes from 12 undulated young emmetropes in 4.0 mm, 3.0 mm, and 2.0 mm pupil sizes with a ray tracing wavefront aberrometer under accommodative stimuli from 0 to 4.00 D. RESULTS: RMS values of total high-order aberrations (HOAs), total coma, total higher-order astigmatism, total spherical, and total trefoil aberrations significantly decreased as pupil size decreased in the relax state. In 4.0 mm, 3.0 mm, and 2.0 mm pupil sizes, the average root mean square (RMS) value of total coma was the highest item of HOAs in the relaxed state. With accommodation up to 3.00 D, the RMS of HOAs remained constant but changed significantly at 4.00 D stimulation (P <0.05) in all these three pupil sizes. The 4th spherical aberration decreased and changed from positive to negative with increasing accommodation over 4.0 mm size, whereas it remained positive under all accommodative levels over the 3.0 mm and 2.0 mm zones, and no trend was found. The 3rd order trefoil 0 changed from negative to positive with increasing accommodation, with 4.0 mm, 3.0 mm, and 2.0 mm pupil sizes. CONCLUSIONS: In 4.0 mm, 3.0 mm, and 2.0 mm pupil sizes, under accommodative stimuli from 0.00 to 4.00 D, dramatical trends amongst the HOAs, spherical, coma, and trefoil aberrations were demonstrated. Most of the higher-order aberrations induced by accommodation deserves further investigation. Simple elimination of the higher-order aberrations will be replaced by the optimization of aberrations.

Long-term visual outcome of dense bilateral congenital cataract.

BACKGROUND: Dense congenital cataracts often cause severe visual impairment. The results of long-term follow-up of dense bilateral congenital cataract in China have not been well documented. The purpose of this study was to evaluate the long-term visual function in children who underwent cataract extraction for dense bilateral congenital cataract in southern part of China. METHODS: Medical records of children who underwent surgery of dense bilateral congenital cataract between January 1992 and December 2000 at Zhongshan Ophthalmic Center of Sun Yat-sen University were retroactively reviewed. In 38 children available for current follow-up, best corrected visual acuity (BCVA) and stereoscopic vision, as well as nystagmus, strabismus, and other complications, were evaluated. The mean follow-up period was 107.6 months (range 60 to 167 months). RESULTS: The mean age of cataract extraction and secondary intraocular lens implantation were 5.6 months (range 3 to 12 months) and 4.2 years (range 2.4 to 15 years), respectively. The mean BCVA was 0.25 in the better eye and 0.16 in the fellow eye. Stereoscopic vision was absent in all patients, and 3 children had simultaneous perception. Nystagmus was detected in all cases and strabismus in 35 cases. A high correlation was found between timing of cataract extraction and final BCVA of the better eye (r = -0.55, P = 0.00). A statistically significant difference was found in BCVA between post- and pre-treatment of amblyopia (t = 5.65, P = 0.00). CONCLUSIONS: Long-term visual function in children with dense bilateral congenital cataract was poor when cataract surgery was performed at age of 3 months or later. Earlier cataract surgery with adequate optical rehabilitation contributed to better visual outcome.

Comparison of higher-order aberrations and contrast sensitivity between Tecnis Z9001 and CeeOn 911A intraocular lenses: a prospective randomized study.

BACKGROUND: The advent of wavefront technology in the past five years has provided some insight into the optical outcomes of cataract surgery. The Tecnis Z9001 intraocular lens (Tecnis IOL, AMO) with a modified prolate anterior surface was claimed to reduce or even eliminate ocular spherical aberration to improve the visual quality. The purpose of this study was to determine whether Tecnis IOLs can improve the quality of vision as evaluated by measuring visual acuity, wavefront aberration, and contrast sensitivity. METHODS: In an intraindividual prospective study, 20 patients with bilateral cataract were randomly assigned to receive a modified prolate anterior surface IOL (Tecnis Z9001, AMO) in one eye and a biconvex spherical surface IOL (CeeOn 911A, AMO) in the other. After 3 months, the following were investigated: best corrected visual acuity, pupil diameter, photopic and mesopic contrast sensitivity, and wavefront aberration of the whole eye (ocular), cornea, and internal plane. RESULTS: The differences in the best corrected visual acuity and pupil diameter between the two groups were not statistically significant. Negative 4th-order spherical aberration (Z(4)(0)) was found in the Tecnis group, whereas positive Z(4)(0) found in the CeeOn group for the internal and ocular plane. Statistically significant differences were found at the ocular higher-aberrations between the two IOLs. Contrast sensitivity testing showed significantly better results in the Tecnis group at visual angles higher than 1.0 degree under photopic conditions and at visual angles higher than 1.6 degree under mesopic conditions both without glare and with glare in comparison with the CeeOn group. CONCLUSIONS: The Tecnis Z9001 IOL with a modified prolate anterior surface produces negative spherical aberration and consequently reduces the higher-order aberrations in pseudophakic eyes. This leads to enhanced contrast sensitivity and improved functional vision compared to conventional spherical IOLs.