[Analysis of genetic diversity of Sichuan indigenous chicken breeds using microsatellite markers].

Thirty microsatellite markers with medium or high polymorphisms were selected to detect the genetic diversity of 8 indigenous chicken breeds in Sichuan. According to the allele frequencies of 30 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and DA genetic distances were calculated for each breeds. The results showed that 24 of 30 microsatellite sites were highly polymorphic, so the 24 microsatellite markers were effective markers for analysis of genetic relationship among chicken breeds. The mean heterozygosity of 8 chicken breeds was all over 0.5. The highest was the Luning chicken (0.681), and the lowest was the Jiuyuan Dark chicken. The high diversity of 8 chicken breeds might be caused by the traffic obstruction(geographic isolation). The results of the heterozygosity were consistent with that of PIC. UPGMA tree was completed through analysis of DA genetic distances. Emei Dark chicken, Miyi chicken, Luning chicken and Jiuyuan Dark chicken were the first group: Miyi chicken and Luning chicken were grouped firstly, then Emei Dark chicken were grouped with them in shorter time distances, and Jiuyuan Dark chicken were grouped with them at last. Shimiancao Ke chicken Xingwen Silky chicken and Muchuan Silky were the second group: Xingwen Silky chicken and Muchuan Silky were grouped firstly, and then Shimiancao Ke chicken was grouped with them. Liangshangya Ying chicken had its own branch. The result of UPGM was consistent with the genesis, breeding history, differentiation and location of 8 chicken breeds.

Modulation of cyclophilin gene expression by N-4-(hydroxyphenyl)retinamide: association with reactive oxygen species generation and apoptosis.

To explore the mechanisms underlying the pro-apoptotic effects of the synthetic retinoid N-4-(hydroxyphenyl)retinamide (4-HPR) on LNCaP human prostate cancer cells, we used the differential display-polymerase chain reaction (DD-PCR) technique to identify 4-HPR-responsive genes. RNA extracted from LNCaP cells that had been treated for 24 h with 4-HPR at a dose (2.5 microM) optimal for apoptosis induction was used for DD-PCR analysis using random primers. A differentially expressed 115 bp fragment was cloned and sequenced and then identified in GenBank as having a high degree of homology with several members of the cyclophilin gene family. Northern blot analyses using specific probes for cyclophilin A, cyclophilin D, and the cloned 115-bp fragment were performed on RNA extracted from LNCaP cells and MCF-7 human breast cancer cells treated with 4-HPR, N-acetylcysteine (NAC, an anti-oxidant), 4-HPR plus NAC, cyclosporin A, R-1881 (a synthetic androgen), dehydroepiandrosterone, all-trans retinoic acid, or prednisone. 4-HPR downregulated the transcript detected by the 115-bp fragment. Expression patterns detected by the 115-bp fragment and cyclophilin D probes were identical in response to each treatment; none of these treatments affected cyclophilin A expression. Furthermore, expression of mRNA transcripts detected by the 115-bp fragment and cyclophilin D probes correlated with the generation of reactive oxygen species (ROS), as detected by measurement of 2,7-dichlorofluorescein oxidation. Therefore, members of the cyclophilin gene family, such as cyclophilin D (a component of the mitochondrial permeability transition pore previously linked with oxidative stress and apoptosis), may play a role in the ROS-mediated apoptotic effects of 4-HPR.