Analysis on the gene mutations of MYOC in primary open angle glaucoma pedigree / 中华检验医学杂志

Objective To screen the mutations of MYOC gene in a Chinese primary open angle glaucoma (POAG) family from Cbengqing and investigate the relationship between the mutations in MYOC/TIGR gene and POAG.Methods In a large 4-generation glaucoma family, myocilin gene (MYOC) was screened in 39 family members, 8 of which were confirmed patients. Normal controls included 100 normal Chinese subjects.The known mutations of MYOC gene ( including G34C, C136T, G144T, G227A, C624G,G736A, C1009G, A1036G, C1081T, G1099A, G1138A, A1139C, T1430A, C1441A and C1442T) were detected by single strand conformation polymorphism(SSCP) , po]ymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and DNA sequencing.Results G227A mutation was detected in 2 POAG patients and 1 asymptomatic patient, but not in the controls.Cl009del mutation was identified in all patients of the pedigree and an offspring member but not in the controls. No other mutations were detected.Since the C1009del mutation was revealed for the first time, a new GenBank number FJ237047 correponding to ACI62293 was applied.Conclusions The G227A mutation is a known site and there is no relationship between G227A mutation and glaucoma. But C1009del may be related to glaucoma which suggests that morbidity could be higher in the relatives of POAG than the controls.

In-cell assembly of single-chain Fv from peripheral blood lymphocytes from vitiligo patients / 中华皮肤科杂志

Objective To obtain in situ pairing of the variable region genes of the immunoglobulin heavy and light chains of B cells from vitiligo patients. Methods Fifty vitiligo patients were screened by a live melanocyte enzyme-linked immunoabsorbent assay. The sera from seven patients were proved to be strongly positive. Then CD19+ B cells were isolated from the peripheral blood lymphocytes of the 7 patients, then fixed in 10% buffered formaldehyde and permeabilised by Proteinase K. The mRNA was amplified within the cells by reverse transcription-polymerase chain reaction (RT-PCR) with specific primers. The immunoglobulin VH and VL DNA assembled within the same cells using the Cre/loxP system. Nested primers were designed to amplify the known major human VH and VL gene familes. Result A unique 800bp band was obtained corresponding in size to single chain Fv fragments. Conclusion The in situ pairing of the variable region genes of the immunoglobulin heavy and light chains of B cells is obtained from vitiligo patients by in-cell PCR.