Evaluating the association between pathological myopia and SNPs in RASGRF1. ACTC1 and GJD2 genes at chromosome 15q14 and 15q25 in a Chinese population.

BACKGROUND: This study investigated the association of the 27 SNPs located in RASGRF1. GJD2, and ACTC1 genes with pathological myopia in a Chinese Han population. METHODS: Myopia patients were stratified according to whether they did (n = 274) or did not (n = 131) have myopic macular degeneration (MMD). The SNPbrowser software was used to identify specific SNPs for analysis and minimal allele frequency of >20%, and a pairwise r(2) < 0.85 were genotyped using MALDI-TOF mass spectrometry. RESULTS: Before controlling for false discovery rate, the frequency of the rs1867315 C/C genotype compared with healthy controls was lower in the myopia group (p = 0.006) and in myopia patients without macular degeneration (p = 0.019). The frequency of the rs670957A/A genotype was also lower in patients without MMD compared with controls (p = 0.045). For rs2070664, the frequency of the A allele was higher in the patients with MMD compared to those without MMD (p = 0.032). After controlling for a false discovery rate of 5%, there was no significant difference in genotype and allele frequencies between these groups. CONCLUSION: In this study, there was no association of the analyzed SNPs located in RASGRF1. GJD2, and ACTC1 with pathological myopia, suggesting that SNPs included in our study have no or a limited role in causing pathologic myopia in this Chinese Han population.

Refractive change in the adult rabbit eye after corneal relaxation with the femtosecond laser.

BACKGROUND: A new procedure to correct myopia that does not disturb the cornea in the optical zone and avoids injuring the corneal epithelium could be a key advance in corneal refractive surgery. The aim of this study is to observe the refractive change in the adult rabbits undergoing femtosecond laser-assisted multilayer intrastromal ablation in the mid-periphery of the cornea without injury of epithelium. METHOD: The right eyes of 8 New Zealand White adult rabbits were used for the experiments. A 60-kHz femtosecond laser delivery system was used, and three lamellar layers of laser pulses were focused starting at a corneal depth of 180 µm and ending at 90 µm from the surface, with each successive layer placed 45 µm anterior to the previous layer. In the interface of the applanation contact lens cone, a 6-mm diameter aluminum circle was placed at the center to block the laser, limiting ablation to the mid-periphery of the cornea. The laser settings were as follows: spot/line separation, 10 µm; diameter, 8.0 mm; energy for ablating the stroma, 1.3 µJ. An authorefractor was used to assess the manifest refraction. RESULTS: Mean spherical equivalent (SE) (mean ± SD, SD: standard deviation) was significantly increased at postoperative week 1 (1.67 ± 0.26 D, p < 0.0001), month 1 (1.65 ± 0.23 D, p < 0.0001), and month 3 (1.60 ± 0.22 D, p < 0.0001) compared to baseline (0.68 ± 0.27 D). Mean spherical equivalent showed no significant change between postoperative week 1 and month 3 (p = 0.1168). CONCLUSION: Femtosecond laser-assisted multilayer corneal intrastromal ablation in the mid-periphery may cause a consequent hyperopic shift with no refractive regression.

[Changes of opsin expression in experimental form-deprivation and defocus myopia in guinea pig].

OBJECTIVE: To investigate the opsin expression in form-deprived and defocus myopia in guinea pig and to study the relationship between the opsin expression and the experimental myopia. METHODS: Fifty guinea pigs were randomized into form-deprived group, defocus group (n = 20 in each group) and normal group (n = 10). Guinea pigs in form-deprived group wore a diffuser RGP on one eye since one week after birth. Those in defocus group wore -4.00 D RGP on one eye. The contralateral eyes wore 0 D RGP were used as the control. Refraction, axial length and depth of vitreous cavity were measured after 1 and 2 weeks. Animals were sacrificed and the retina were dissected at 10:00 - 12:00 AM. The level of opsin and its mRNA were measured by Western-blot and real-time PCR, respectively. RESULTS: Two weeks after the experiment, the refraction in form-deprived group and defocus group were (-4.00 ± 0.87) and (-2.00 ± 1.17) D respectively, which were significant different compared with contralateral eyes or normal control group (F = 203.98, 88.66, P < 0.05). These also accompany with increase of axial length and depth of vitreous cavity in form-deprived group and defocus group. Expression of S-opsin mRNA were increased both in form-deprived and defocus groups and the ratios of S-opsin mRNA/ß-actin expression were 0.752 ± 0.05 and 1.117 ± 0.13 in two weeks treatment, which were significant different from contralateral eyes (the ratios of S-opsin mRNA/ß-actin expression were 0.536 ± 0.04 and 0.772 ± 0.10.t = 6.10, 6.28, P < 0.05). Similar findings were also demonstrated in the expression of L-opsin mRNA, which were increased in form-deprived group and defocus group (the ratios of L-opsin mRNA/ß-actin expression were 0.42 ± 0.01) compared with contralateral eyes (the ratios were 0.24 ± 0.0 and 0.34 ± 0.04. t = 6.30, 4.93, P < 0.05) after two weeks experiment. The western-blot results also indicated the high expression of S-opsin and L-opsin level compared to contralateral eyes in form-deprived and defocus groups. CONCLUSIONS: Cone might be the detector receiving the signal of form-deprivation and defocus. Changes of opsin expression might play a role in the occurrence of experimental myopia in guinea pig.

[Stromal cells change after epipolis laser in situ keratomileusis and laser-assisted subepithelial keratectomy in rabbits].

OBJECTIVE: To compare the effects of epipolis laser in situ keratomileusis (Epi-LASIK) and laser-assisted subepithelial keratectomy (LASEK) on the corneal stromal cells, and to evaluate their effects on corneal haze histopathologically. METHODS: Forty-eight New Zealand white rabbits (96 eyes) were randomly treated with Epi-LASIK in one eye and LASEK in the other, and 2 rabbits (4 eyes) without any treatment were used as control. Cornea stromal cell apoptosis was evaluated by terminal deoxyribonucleotidyl transferase-mediated deoxynuridine triphosphate nick end labeling (TUNEL) assay. Stromal cells proliferation and myofibroblasts generation were evaluated by immunocytochemical analyses the expression of Ki-67 and alpha-smooth muscle actin (alpha-SMA) respectively. Myofibroblast generation was further testified by Western blot analysis of alpha-SMA. Both number of TUNEL, Ki-67 and alpha-SMA positive cells and expression of alpha-SMA were semi quantitatively analyzed to explore their effects on corneal haze. RESULTS: Many TUNEL-positive cells appeared in the central anterior stromal in early stages after both LASEK and Epi-LASIK, and the number of TUNEL-positive cells reached a peak 24 hours after either LASEK or Epi-LASIK. There were more TUNEL positive cells found in LASEK group than that in Epi-LASIK group in 1 week postoperatively (t = 3.63, 7.80, 4.34, 2.95, all P < 0.01). There were a lot of Ki-67-positive cells in anterior corneal stroma after both LASEK and Epi-LASIK, but the peak appeared at 72 hours after the treatments. The significant difference was also found in the number of Ki-67-positive cell between the two treatments in 1 week postoperatively (t = 3.81, 5.85, 5.09, 5.59, all P < 0.01). alpha-SMA-positive cells started to appear apparently at 1 week after both LASEK and Epi-LASIK, and both the peaks appeared at 1 month after the treatments, and there were still a lot of alpha-SMA-positive cells in corneal stroma at 3 months after LASEK. There were more alpha-SMA-positive cells found after 1 week in LASEK group than that in Epi-LASIK group (t = 2.97, 7.19, 6.73, all P < 0.01). CONCLUSION: There are less stromal cell apoptosis, proliferation and myofibroblast generation in Epi-LASIK than that in LASEK. Therefore, comparing with LASEK, Epi-LASIK induces less corneal haze response in correcting myopia.

[Clinical investigation of epipolis laser in situ keratomileusis on high myopia].

OBJECTIVE: To investigate the surgical characteristics and clinical results of Epi-LASIK on high myopia. METHODS: Epi-LASIK was performed with a rotational epikeratome, which features a blunt oscillating blade. Corneal epithelial separation was achieved mechanically without the use of alcohol. Laser ablation was performed with Zeiss Mel-80 excimer laser. After ablation, epithelial sheet was repositioned and a therapeutic contact lens was applied to the eye for 3 to 5 days. The postoperative symptom, epithelial flap, contrast sensitivity, wavefront aberration and haze formation were investigated. The period of follow-up ranged from 3 to 6 months. RESULTS: 52 eyes of 30 patients with a mean spherical equivalent (SE) of (-12.82 +/- 4.38) D were included in the clinical trial. Among which 36 eyes were pathologic myopia and 16 eyes were simple high myopia. Patient age ranged from 20 to 50 years. Integrated epithelial flap with a diameter of 8 to 9 mm was made in 49 eyes leaving an about 3 mm superior hinge. Epithelial flap contained part of stroma tissue was seen in one eye. Free epithelial flap was made in 2 eyes. 4 eyes had moderate photophobia, tearing and foreign body sensation. Other patients only had mild discomfort. No patient reported other subjective symptom after 2 days with the exception of mild foreign body sensation. In 3 months after surgery, spherical equivalent of 44 eyes (84.61) were within +/- 1.00 D of attempted refraction. No eye lost more than one line of best spectacle-corrected visual acuity (BSCVA), whereas 10 eyes (19.23%) gained one or two lines of BSCVA. Grade 0.5 haze was found in 8 eyes (15.38%) and grade 1 haze was found in 2 eyes (3.84%). In 6 months after surgery, Grade 0.5 haze was found in 3 eyes. Contrast sensitivity decreased 1 month postoperatively and recovered to preoperative lever gradually 3 months postoperatively. Higher order aberration was increased postoperatively. CONCLUSIONS: Epi-LASIK is a safe and efficient method to correct high myopia with the advantages of having mild irritating symptom and haze. Larger series with adequate follow-up are needed to observe the long term clinical results and haze.