Transcriptome analysis of sheep follicular development during prerecruitment, dominant, and mature stages after FSH superstimulation.

Sheep is usually a monovular animal; superovulation technology is used to increase the number of offspring per individual and shorten generation intervals. To date, mature FSH superstimulatory treatments have been successfully used in sheep breeding, but much remains unknown about genes, pathways, and biological functions involved in follicular development. Therefore, in this study, we performed transcriptome profiling of small follicles (SFs; 2-2.5 mm), medium follicles (MFs; 3.5-4.5 mm), and large follicles (LFs; > 6 mm) in Mongolian ewes after FSH superstimulation. Furthermore, we identified differentially expressed genes and performed Kyoto Encyclopedia of Genes and Genomes pathway and Gene Ontology enrichment analyses in 3 separate pairwise comparisons. We found that ovarian steroidogenesis was significantly enriched in the SFs versus MFs analysis; the associated genes, cytochrome P450 family 19 (CYP19) and Hydroxy-delta-5-steroid dehydrogenase 3 beta- and steroid delta-isomerase 1 (HSD3B1), were significantly upregulated. Moreover, proline metabolism, glutathione metabolism, and PPAR signaling pathways were significantly enriched in the LFs versus SFs analysis; the associated genes, glutamate-cysteine ligase modifier subunit (GCLM) and cystathionine gamma-lyase (CTH), were significantly upregulated, whereas peroxisome proliferator-activated receptor gamma (PPARγ) was significantly downregulated. In summary, our study provides basic data and possible biological direction to further explore the molecular mechanism of sheep follicular development after FSH superstimulation.

Alkaloids from Cynanchum komarovii with inhibitory activity against the tobacco mosaic virus.

A pyrroloisoquinoline alkaloid, 2,3-dimethoxy-6-(3-oxo-butyl)-7,9,10,11,11a,12-hexahydrobenzo[f]pyrrolo[1,2-b]isoquinoline (1), whose structure was determined by spectroscopic methods, was isolated from the aerial parts of Cynanchum komarovii, together with two known alkaloids, 7-demethoxytylophorine (2) and 7-demethoxytylophorine N-oxide (3). Alkaloids 2 and 3 had antiviral activities against tobacco mosaic virus.

[Expression of Trial in mammalian cell and its apoptosis activation in culture cells].

To develop a system for expression of Trail in mammalian cell. PCR and sequencing were used to clone and to confirm Trail gene. Then, we constructed the vector for expression of Trail in mammalian cells. HeLa cells, one of sensitive cell lines to Trail, were transfected with the vector DNA. After 48 h, flow cytometry was used to screen HeLa transfected by Trail. 19% of total cells happened to apoptosis. This study conforms that the production of Trail expressing in mammal cell can induce apoptosis and exhibit high bio-active. The results indicate the potential application of Trail to tumor immunotherapy.

[Expression of 3'-truncated preS/S gene in transgenic mice].

The 3'-truncated preS/S region from HBV genome, encoding a transcriptional transactivator, was cloned and a recombination expression vector for 3'-truncated HBV preS/S sequences under the control of the CMV promoter was constructed by recombination DNA techniques. Then, the expression vector DNA was microinjected into pronuclei of fertilized mouse oocytes. Founders of transgenic mice harbouring the recombination gene which can be expressed were screened by polymerase chain reaction (PCR) at genomic DNA level and confirmed by ELISA andlysis at protein level. Two of 15 mice in one series of microinjections showed the expression of 3'-truncated preS/S gene from HBV genome. The expression vector of 3'-truncated preS/S gene might be helpful for further studies of relationship between the expression production of 3'-truncated preS/S sequence and HBV-associated oncogenesis in vitro. The transgenic mice with expressing 3'-truncated preS/S gene will provide new insight into its roles in the development of human hepatocelluar carcinoma (HCC) in vivo.