Construction of human Bcl-6 3'UTR reporter vector and expression vector and their functional assessment / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the direct regulation of miR-127 on Bcl-6 and the effect of Bcl-6 in rescuing miR-127-induced cell cycle and cell growth inhibition.</p><p><b>METHODS</b>The 3'UTR and coding region of human bcl-6 gene were amplified by PCR and cloned into pcDNA3.0-Luc and pcDNA3.0-Flag vectors, respectively. Mutations were introduced into the seed sequences of the predicted miR-127 target sites within the Bcl-6 3'UTR using recombinant PCR. Luciferase assay was used to verify the direct targeted regulation of miR-127 on Bcl-6. In HepG2 cell models with overexpression or knockdown of miR-12, the changes of cell cycle and cell growth were investigated after transfection with the constructed vectors.</p><p><b>RESULTS</b>The recombinant plasmids were successfully obtained as confirmed by double digestion and sequence identification. Luciferase assay showed that in 293T and HepG2 cells, miR-127 inhibited the activation of wild-type Bcl-6 3'UTR reporter vector but not mutated Bcl-6 3'UTR vector. Overexpression of miR-127 induced cell cycle arrest at G(2)/M phase and suppressed the growth of HepG2 cells, and these effects were reversed by Bcl-6 overexpression.</p><p><b>CONCLUSION</b>We successfully cloned wild-type and mutated 3'UTR reporter vectors and expression vector of bcl-6 gene and confirmed their biological functions.</p>

Clinical features and confocal microscopic imaging characteristics of 466 cases with infectious keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the role of confocal microscopy in infectious keratitis management. METHODS:Totally 466 patients (467 eyes) diagnosed as infectious keratitis from January 2010 to December 2013 were retrospectively studied. the corneas were examined early by in vivo confocal microscopy. The characteristics of their images and clinical features were studied and summarized. RESULTS:All patients were recorded, the average age was 54. 4±13. 0 years, in which 264 cases (56. 7%) were male, and 202 cases ( 43. 3%) were female. In the 466 patients, 190 (40. 8%) were fungal keratitis, 148 (31. 8%) were viral keratitis, 125 (26. 8%) were bacterial keratitis and 3 ( 0.6%) were acanthamoeba keratitis. There were fungal hyphae in the images of fungal keratitis. Amebic cysts were found in acanthamoeba keratitis. CONCLUSION:Confocal microscope can help the early diagnose and treatment of infectious keratitis. It is a noninvasive imaging technique that provides high resolution images of ocular structures at a cellular level and infectious keratitis represents one of its most important clinical uses.

Changes of retinal nerve fiber layer thickness by optic coherence tomography in the early stage of diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate changes of retinal nerve fiber layer ( RNFL) thickness and macular retinal thickness in patients with early diabetic retinopathy ( DR ) and disclose the changing trends of RNFL thickness and macular retinal thickness in different stages of early DR. METHODS:It was a clinical case control study. Through selecting 60 patients ( 120 eyes ) with early DR diagnosed with type 2 diabetes were divided into non - diabetic retinopathy (NDR) group (20 cases, 40 eyes) and mild non-proliferative diabetic retinopathy (NPDR) group (20 cases, 40eyes), moderate NPDR group (20 cases, 40 eyes) . Twenty normal patients ( 40 eyes ) were enrolled as control group. The RNFL thickness of optical nerve ( with circle scan round optic nerve head, scan diameter of 3. 45mm) and the retinal thickness of macular ( around center point with 1000μm diameter ) were measured by optical coherence tomography ( OCT ) , to compare the RNFL thickness changes of the control and early DR patients. RESULTS:Compared with the control group, the RNFL thickness of optical disc in the inferior quadrant was descended obviously in NDR group ( P 0.05). In mild NPDR group, the RNFL thickness of optical disc in the mean and inferior quadrant was significantly descended than that in the NDR group. In moderate NPDR group, the RNFL thickness of optical disc in the mean, superior and inferior quadrant was statistical significance descended than that in the NDR group ( P CONCLUSION:With the development in the degree of early DR, RNFL thickness is gradually decreased and retinal thickness is increased, OCT can be observed qualitatively and quantitatively in DR.