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The complete mitochondrial genome for Gymnocypris eckloni and Gymnocypris przewalskii ganzihonensis is 16,686 and 16,682 bp in size, respectively. Both of the mitochondrial genomes have identical genomic organization and structure including 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes and 1 control region as found in other vertebrates, with the exception of 86 bp non-coding nucleotides between tRNA Thr and tRNA Pro. The phylogenetic analyses based on the concatenated nucleotide sequence of 12 PCGs on the heavy strand supported a relatively closed relationship among G. eckloni, G. p. przewalskii and G. p. ganzihonensis. The sequence divergence between G. eckloni and G. przewalskii, as well as within G. przewalskii, is relatively low compared to the average sequence divergence among other cyprinid fishes, suggesting that they are the most closely related species resulted from a relatively recent speciation event. The estimated molecular divergence time is coincided with a relatively recent speciation event occurring during the separation of the Yellow River and Qinghai Lake. [ABSTRACT FROM AUTHOR]

Diabetic retinopathy (DR) is a major cause of blindness, and there is a lack of effective treatment at present. Rho-associated coiled-coil containing serine/threonine protein kinases (ROCKs) have recently been suggested as potential targets for the DR treatment. We herein report the discovery of 4 H -chromen-4-one derivatives as a new class of ROCK inhibitors. Structure-activity relationship analyses led to the identification of the most active compound, 4-(dimethylamino)- N -(3-{2-[(4-oxo-4 H -chromen-7-yl)oxy]acetamido}phenyl) ( 12j ). This compound showed excellent kinase selectivity for ROCK I and ROCK II against 387 other kinases. In retinal explants, compound 12j protected retinal neurons from high glucose-induced oxidative stress and apoptosis-mediated cell death. Furthermore, 12j administration suppressed the improper proliferation of Müller cells and promoted the regression of vascular vessels in retinal explants cultured in a high glucose microenvironment. Collectively, our data suggest that 12j could be a potential lead compound for the treatment of DR, hence deserving further in-depth studies.

The full-length cDNA of hTIMP-1 was obtained from a surgical patient with HCC by the method of RT-PCR. Then it was cloned into the adenoviral shuttle plasmid pAdTrack-CMV, and subsequently cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thereupon, a recombinant adenoviral plasmid containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. The adenoviruses (AdhTIMP-1) were packaged and amplified in adenoviral packaging cells HEK 293. Then the viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 was detected by the techniques of Western blot and RT-PCR. The recombinant adenovirus vector carrying human TIMP-1 was successfully constructed and expressed in vitro and may pave the way for further application in liver gene therapy.

The fragment of MDR1 gene obtained from the plasmid pHaMDR1-1 carrying the whole human MDR1 cDNA, was cloned reversely into the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the Escherichia coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged in the 293 cells. The recombinant adenovirus MDR1 vector would introduce the antisense MDR1 gene into the human multidrug resistance hepatocellular cell line effectively, which would provide an experimental basis for studies on the multidrug resistance in human hepatocellular carcinoma.