Interfering RNA against PKC-α inhibits TNF-α-induced IP3R1 expression and improves glomerular filtration rate in rats with fulminant hepatic failure.

We have reported that tumor necrosis factor-α (TNF-α) is critical for reduction of glomerular filtration rate (GFR) in rats with fulminant hepatic failure (FHF). The present study aims to evaluate the underlying mechanisms of decreased GFR during acute hepatic failure. Rats with FHF induced by d-galactosamine plus lipopolysaccharide (GalN/LPS) were injected intravenously with recombinant lentivirus harboring short hairpin RNA against the protein kinase C-α ( PKC-α) gene (Lenti-shRNA-PKC-α). GFR, serum levels of aminotransferases, creatinine, urea nitrogen, potassium, sodium, chloride, TNF-α, and endothelin-1 (ET-1), as well as type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) expression in renal tissue were assessed. The effects of PKC-α silencing on TNF-α-induced IP3R1, specificity protein 1 (SP-1), and c-Jun NH2-terminal kinase (JNK) expression, as well as cytosolic calcium content were determined in glomerular mesangial cell (GMCs) with RNAi against PKC-α. Renal IP3R1 overexpression was abrogated by pre-treatment with Lenti-shRNA-PKC-α. The PKC-α silence significantly improved the compromised GFR, reduced Cr levels, and reversed the decrease in glomerular inulin space and the increase in glomerular calcium content in GalN/LPS-exposed rats. TNF-α treatment increased expression of PKC-α, IP3R1, specificity protein 1 (SP-1), JNK, and p-JNK in GMCs and increased Ca2 + release and binding activity of SP-1 to the IP3R1 promoter. These effects were blocked by transfection of siRNA against the PKC-α gene, and the PKC-α gene silence also restored cytosolic Ca2+ concentration. RNAi targeting PKC-α inhibited TNF-α-induced IP3R1 overexpression and in turn improved compromised GFR in the development of acute kidney injury during FHF in rats.

Tumor necrosis factor-alpha-induced reduction of glomerular filtration rate in rats with fulminant hepatic failure.

The mechanism of renal failure during fulminant hepatic failure (FHF) or end-stage of liver disease is not fully understood. The present study aims to delineate the mechanisms of decreased glomerular filtration rate (GFR) in acute hepatic failure. A rat model of renal insufficiency in severe liver injury was established by lipopolysaccharide (LPS) plus D-galactosamine (GalN) exposure. GFR was evaluated by continuous infusion of fluorescein isothiocyanate-inulin with implanted micro-osmotic pumps. GalN/LPS intoxication resulted in severe hepatocyte toxicity as evidenced by liver histology and biochemical tests, whereas renal morphology remained normal. GFR was reduced by 33% of the controls 12 h after GalN/LPS exposure, accompanied with a decreased serum sodium levels, a marked increase in serum TNF-α and ET-1 levels as well as significantly upregulated renal type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) expression. The upregulated IP3R1 expression was abrogated by the treatment of anti-TNF-α antibodies, but not by 2-aminoethoxydiphenylborate (2-APB), which blocks the inositol 1,4,5-trisphosphate signaling pathway. Treatments with either TNF-α antibodies or 2-APB also significantly improved the compromised GFR, elevated serum urea nitrogen and creatinine levels, and reversed the decrease in glomerular inulin space and the increase in glomerular calcium content in GalN/LPS-exposed rats. The extent of acute liver injury as reflected by serum ALT levels was much more attenuated by anti-TNF-α antibodies than by 2-APB. Liver histology further confirmed that anti-TNF-α antibodies conferred better protection than 2-APB in GalN/LPS-exposed rats. LPS-elicited TNF-α over-production is responsible for decreased GFR through IP3R1 overexpression, and the compromised GFR resulted in the development of acute renal failure in rats with FHF.

Graphite prelithiation by solid electrochemical corrosion of lithium metal with a superficial mosaic structure.

A transformative concept of solid electrochemical corrosion has been put forward, in which solid-state electrolyte LiPON has been applied to replace the liquid one to prelithiate graphite with Li-metal. Thus, high prelithiation efficiency and low polarization of the treated anode can be obtained, with a unique mosaic structure left at the surface.

[Analysis of the genetic structure of 12 Chinese and foreign cattle breeds using DNA microsatellite markers].

The genetic diversity of 9 Chinese native cattle breeds and 3 introduced breeds were analyzed using 12 microsatellite DNA markers recommended by the Food and Agriculture Organization of the United Nations (FAO) and the International Society of Animal Genetics (ISAG) through fluorescence-multiple PCR. According to the allele frequencies of 12 microsatellite locus, mean heterozygosity (H), polymorphism information content (PIC) , DA and DS genetic distances were calculated for each breed. Cluster analysis based on UPGMA method indicated that 12 breeds were clustered into four groups. Group I belonged to the southern-China cattle including Enshi, Liping, Zhaotong and Chuannan Mountainous; Group II belonged to the central-China cattle including Jiaxian Red and Zaosheng, Pinglu Mountainous; Group III belonged to the northern-China cattle including Yanbian and Changbai; Group IV consisted of foreign breeds including Germany Yellow, Simmental and Charolais. The results may provide an academic basis for preservation and utilization of Chinese native cattle breeds.

[Analysis on the genetic structure of 8 Asia buffalo populations].

One hundred and forty seven alleles were detected when thirteen microsatellite loci were analyzed applying fluorescence-multiplex PCR technology in eight buffalo populations were analyzed, including six indigenous Chinese native breeds (DechangXinglongFuzhongWenzhouDongliuFu'an), and two introduced breeds (MurrahNili-Ravi). Seven populations have their own unique alleles, total number is twenty-three. As to all the eight populations, effective number of alleles (Ne) was between 2.2908 and 4.2308, heterozygosity (H) between 0.4951 and 0.7194, and polymorphism information content (PIC) between 0.4495 and 0.6776. Eleven of the thirteen microsatellite loci were of high polymorphism and were then the appropriate, polymorphism marker could be used to analyze properly genetic diversity of the involved buffalo populations. Cluster analysis indicated that Fuzhong and Dongliu were clustered together, then with an independent cluster of Xinglong. Wenzhou and Fu'an were clustered together, Dechang was an independent cluster. Murrah and Nili-Ravi were clustered together.