A novel CRX mutation by whole-exome sequencing in an autosomal dominant cone-rod dystrophy pedigree.

AIM: To identify the disease-causing gene mutation in a Chinese pedigree with autosomal dominant cone-rod dystrophy (adCORD). METHODS: A southern Chinese adCORD pedigree including 9 affected individuals was studied. Whole-exome sequencing (WES), coupling the Agilent whole-exome capture system to the Illumina HiSeq 2000 DNA sequencing platform was used to search the specific gene mutation in 3 affected family members and 1 unaffected member. After a suggested variant was found through the data analysis, the putative mutation was validated by Sanger DNA sequencing of samples from all available family members. RESULTS: The results of both WES and Sanger sequencing revealed a novel nonsense mutation c.C766T (p.Q256X) within exon 5 of CRX gene which was pathogenic for adCORD in this family. The mutation could affect photoreceptor-specific gene expression with a dominant-negative effect and resulted in loss of the OTX tail, thus the mutant protein occupies the CRX-binding site in target promoters without establishing an interaction and, consequently, may block transactivation. CONCLUSION: All modes of Mendelian inheritance in CORD have been observed, and genetic heterogeneity is a hallmark of CORD. Therefore, conventional genetic diagnosis of CORD would be time-consuming and labor-intensive. Our study indicated the robustness and cost-effectiveness of WES in the genetic diagnosis of CORD.

[Quality evaluation of Astragali Radix through chemical pattern recognition of fingerprint by HPLC-dAD-ELSD].

OBJECTIVE: To develop an HPLC-DAD-ELSD method for detecting the fingerprint of Astragali Radix and evaluate the quality through similarity calculation and chemical pattern recognition. METHOD: Separation was performed at 25 degreeC on an Agilent Zorbax ODS C18 column(4.6 mm x250 mm,5 microm). Gradient elution was performed with the mobile phases of acetonitrile and water containing 0. 2% formic acid. The flow rate was 0. 8 mL min-1 , and sample size was 10 microL. The UV detection wavelength was set at 280 nm. The drift tube temperature for ELSD was set at 110 degreeC , and the nebulizing gas flow rate was 3.0 L min-1. The similarity calculation and chemical pattern recognition were used for fingerprint analysis. RESULT: The HPLC-DAD-ELSD method for chromatographic fingerprint of Astragali Radix showed better results of stability, precision and repeatability. The reference chromatographic fingerprint of Astragali Radix was established on the eighteen Astragali Radix samples from different sources. The results of similarity calculation were higher than 0. 83, which was in accordance with the result of chemical pattern recognition analysis. CONCLUSION: Fingerprint and chemical pattern recognition analysis could effectively distinguish Astragali Radix from different source, which could be applied to the quality control of Astragali Radix.

[Clinical application of deep lamellar keratoplasty for corneal macula after viral keratitis].

OBJECTIVE: To explore the clinical safety and efficacy of deep lamellar keratoplasty for corneal macula after viral keratitis. METHODS: A total of 16 patients (16 eyes) of corneal macula after viral keratitis at our center from January 2009 to June 2011 received anti-virus eye-drops and then underwent deep lamellar keratoplasty when inflammation were dissolved or quiescent. And the corneal graft diaphaneity and corneal graft rejection reactions were observed. RESULTS: All 16 patients were recovered with a success rate of 100%. After a follow-up of 18-24 months, all corneal grafts were transparent and best-corrected visual acuity was 0.2-0.6. There was no reoccurrence during the follow-up period. After a follow-up of several months, 2 of them had rejection reaction and were controlled after treatment. No rejection reaction occurred during the long-term follow-up. CONCLUSION: As a safe and efficacious procedure, deep lamellar keratoplasty can significantly raise vision acuity with few complications.

[The visual functional therapeutic effects of deep lamellar keratoplasty and penetrating keratoplasty for fungal corneal ulcer].

OBJECTIVE: To investigate the visual functional therapeutic effects of deep lamellar keratoplasty (DLK) and penetrating keratoplasty (PK) on perforated fungal keratitis. METHODS: 64 patients (64 eyes) of fungal corneal ulcer in Ophthalmology Center of Ningbo Yinzhou People Hospital from 2004 to 2009 were retrospected, of which undergo DLK (36 patients, 36 eyes) and PK (28 patients, 28 eyes), and followed up by 12 to 24 months. Check two sets of patients' VA and refraction before operation, and analyze the best-corrected visual acuity (BCVA), corneal refraction changes, corneal endothelium counting and complications, using χ(2) test and t-test. RESULTS: The BCVA after the operation of two sets are both improved, 32 eyes of DLK set were above 0.4, 19 eyes of PK set were above 0.4, of which the DKL set is a bit better than PK set (χ(2) = 4.304, P > 0.05). The astigmatism of DLK set is smaller than the PK set after operation, and there is significant difference (χ(2) = 4.98, P < 0.05). The astigmatism of two sets on the average were all no more than 5.00 D. The reject reaction of keratoplasty of DLK is less than PK, there is obviously significant difference (χ(2) = 34.17, P < 0.05). CONCLUSIONS: There is less complications of DLK than PK for fungal corneal ulcer. DLK can reduce the occurrence of reject reaction of endothelium type, and the BCVA, refraction of DLK after operation is similar to PK, the incidence rate of surgical operation failure is low.