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Cinobufacini, a traditional Chinese medicine, has been used widely for cancer treatment, such as hepatocellular carcinoma (HCC), sarcoma, and leukemia. Previous studies done by our lab indicated that cinobufacini could suppress HCC cells through mitochondria-mediated and Fas-mediated apoptotic pathways. Here, we use a combination of cinobufacini and doxorubicin to inhibit the growth of HCC cells. The combination group induced more significant apoptosis by affecting proteins and RNA of apoptosis-related elements, such as Bcl-2, Bax, Bid, and cytochrome c. Furthermore, cinobufacini, as a mixture of a number of components, had stronger apoptosis-inducing activity than particular individual components or a simple mixture of a few components. Overall, these results suggested that the combination of cinobufacini and doxorubicin may provide a new strategy for inhibiting the proliferation of HCC cells.

The aim of this study was to analyze the relationship between polymorphism of the MHC-DRB1/DQB1 gene and its resistance to Cystic Echinococcosis (C.E), as well as to screen out the molecular genetic marker of antiechinococcosis in Chinese Merino sheep. The MHCII-DRB1/DQB1 exon 2 was amplified by polymerase chain reaction (PCR) from DNA samples of healthy and hydatidosis sheep. PCR products were characterized by restriction fragment length polymorphism (RFLP) technique. Five restriction enzymes (Mval, HaeIII, SacI, SacII, and Hin1I) were employed to cut DRB1, while seven restriction enzymes (MroxI, ScaI, SacII, NciI, TaqI, Mval, and HaeIII) were employed to cut DQB1.Results showed that frequencies of patterns Mvalbb (P < 0.01), SacIab in DRB1 exon 2 (P < 0.05), and TaqIaa, HaeIIInn (P < 0.01) in DQB1 exon 2 were significantly higher in the healthy group compared with the C.E individuals, which implied that there was a strong association between these genotypes and hydatidosis resistance or susceptibility. Chi-square test showed that individuals with the genic haplotype DRB1-SacIab/DRB1-Mvalbb/DQB1-TaqIaa/DQB1-HaeIIInn (P < 0.01) were relatively resistant to C.E, while individuals with the genic haplotypes DRB1-Mvalbc/DQB1-Mvalyy/DQB1-TaqIab/DQB1-HaeIIImn (P < 0.01) and DRB1-Mvalbb/DQB1-Mvalcc/DQB1-TaqIab/DQB1-HaeIIImn (P < 0.01) were more susceptible to C.E. In addition, to confirm these results, a fielding experiment was performed with Chinese Merino sheep which were artificially infected with E.g. The result was in accordance with the results of the first study. In conclusion, MHC-DRB1/DQB1 exon 2 plays an important role as resistant to C.E in Chinese Merino sheep. In addition, the molecular genetic marker of antiechinococcosis (DRB1-SacIab/DRB1-Mvalbb/DQB1-TaqIaa/DQB1-HaeIIInn) was screened out in Chinese Merino sheep.

ABSTRACT. The role of microRNAs (miRNAs) in the regulation of mammalian reproduction has been demonstrated previously. However, only a few studies have assessed the role of miRNAs in the reproduction processes of sheep. The elucidation of miRNA expression profiles in the ovaries of different sheep breeds representing fecundity extremes will be useful in understanding the roles of miRNAs in sheep reproduction. In this study, two small RNA libraries were constructed from ovary tissue taken from Qira black sheep and Hetian sheep during the estrous period and then sequenced using the Solexa sequencing method. We obtained 9,565,212 and 9,563,426 high-quality reads from Qira black sheep and Hetian sheep, respectively. In total, 531 miRNAs, including 98 putative miRNAs, were identified. Among the conserved miRNAs, 125 known miRNAs were significantly differentially expressed in the Qira black sheep and Hetian sheep libraries, with 24 upregulated and 101 downregulated in the Hetian sheep compared to the Qira black sheep. Four differentially expressed miRNAs were analyzed using real-time 7357 miRNA expression analysis in sheep ovaries ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (3): 7356-7367 (2015) quantitative PCR to validate the reliability of the Solexa sequencing results. These results provide a foundation for future research on the regulation of miRNAs in sheep fertility and enrich the sheep miRNA databases.

For this study, peripheral blood samples were collected from human volunteers. Mononuclear cells (MNC) were separated by density centrifugation and were induced to differentiate into endothelial progenitor cells (EPCs) in vitro. Different concentrations of rapamycin and silymarin were introduced to the EPCs over 24 hours and then EPCs were analyzed for proliferation, migration, apoptosis and angiogenesis. Compared with the control group, rapamycin (1, 10, 100 ng/mL) inhibited the proliferation and migration of EPCs in a concentration dependent manner (P<0.05). Silymarin (50, 100 µg/mL) enhanced the proliferation and migration of EPCs and inhibited apoptosis in a concentration dependent manner (P<0.05). By adding rapamycin (1 ng/mL) and silymarin (25, 50, 100 µg/mL) over 24 hours, silymarin inhibited the pro-apoptotic effect of rapamycin on EPCs, and reversed the inhibition of proliferation, migration and angiogenesis of EPCs by rapamycin (P<0.05).

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