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·Long non-coding RNAs (lncRNAs) refer to a kind of non-coding RNA which is longer than 200 nucleotides, with the characteristic of its numerous, diversity of types and modes of action. The biological functions of lncRNA involved in genomic imprinting, chromatin remodeling, translational control of mRNA, cell cycle and cell differentiation control, immune surveillance, constituting the skeleton of nuclear sub structure, etc. LncRNA plays an important role in individual development and human diseases. This paper mainly reviewed those lncRNAs that have been published, and closely related to eye development and diseases.
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AIM: To compare fungal culture and in vivo confocal microscopy ( IVCM ) in the diagnosis of non- primary fungal keratitis.?METHODS:The clinical data of 31 cases (31 eyes) with non- primary fungaI keratitis from September 2016 to February 2017 in our HospitaI were retrospectiveIy reviewed. The positive rate of the two methods was compared by chi-square test.?RESULTS: The positive rate by fungal culture was 58％(18/31 ) and IVCM was 19％ ( 6/31 ); the positive rate comparison difference was statistically significant between fungal culture and IVCM (x2=7. 56,P<0. 01). In the 13 eyes with positive culture results, 2 eyes were positive by IVCM;in the 25 positive IVCM eyes, 14 eyes were positive in culture.?CONCLUSION: The positive rate of fungal culture in non-primary fungal keratitis is higher than that of IVCM. Fungal culture is an essential auxiliary examination in the diagnosis of non - primary fungal keratitis. With the characteristics of fast, noninvasive and repeatable, IVCM also plays an important role in the diagnosis of non-primary fungal keratitis. The combination of the two methods can improve the positive rate of diagnosis.
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<p><b>OBJECTIVE</b>To mutate human annexin V gene and transform it to Pichia Pastoris for mutant human annexin V expression, so as to be purified as active annexin V with endogenous metal chelating site.</p><p><b>METHODS</b>The 5' and 3' end of native annexin V gene were mutated by specific primers. The mutant annexin V gene was inserted into pPIC9K and sequenced. The correct plasmid was linearized and transformed into Pichia Pastoris strain GS115 by electroporation. The transformants were selected from MD plates and cultured in BMGY medium and induced with methanol. The culture was centrifuged and the supernatant was analyzed by SDS-PAGE and silver staining. The binding activity of mutant human annexin V from culture supernatant was determined with phosphatidylserine exposed erythrocytes and fluorescein isothiocyanate-annexin V.</p><p><b>RESULTS</b>The 5' end of native human annexin V gene was fused with GCAGGCGGCTGCGGCCAT coding sequence and 3' end 946-948 site TGT was mutated to AGC. Pichia Pastoris transformants secreted proteins of relative molecular mass 36 000 48 h after methanol induction. The concentration of this protein that inhibited 50% of the binding of fluorescein-annexin V was 4nmol/L.</p><p><b>CONCLUSION</b>Highly-active recombinant mutant human annexin V with endogenous metal-chelating sites can be expressed in Pichia Pastoris system.</p>