In the present work, we cloned the full-length cDNA of the pig Bmi1 gene (BMI1 polycomb ring finger oncogene), which has been indicated as an intestinal epithelial stem cell (IESC) marker in other mammals. This paper provides the first report of the function of Bmi1 in pig intestinal epithelial cells and a brief description of its underlying mechanism. Rapid amplification of cDNA ends technology was used to clone the complete pig Bmi1 sequence, and a Bmi1-pcDNA3.1 vector was constructed for transfection into an intestinal porcine epithelial cell line (IPEC-1). The proliferation ability of the cells was estimated using the MTT assay and the EdU incorporation method at different time points after seeding. Cell cycle information was detected by flow cytometry. The mRNA abundances of cell cycle-related genes were also measured. The results indicated that the pig Bmi1 cDNA is 3,193 bp in length and consists of a 981 bp open reading frame, a 256 bp 5´ untranslated region (UTR), and a 1,956 bp 3' UTR. The transcript contains no signal peptides, and there are no transmembrane regions in the pig Bmi1 coded protein, which has a total of 326 AA. The overexpression of the pig Bmi1 in the IPEC-1 cells led to increased cell proliferation and a lower percentage of cells in the G1 and S phases (P < 0.05), along with a higher percentage of cells in the G2 phase (P < 0.05). Furthermore, the gene expression levels of PCNA, Cyclin D1, Cyclin D2, Cyclin B, CDK1, and CDK2 were all elevated (P < 0.05) by Bmi1 overexpression, while the gene expression levels of Cyclin A2 and p21 showed little difference (P > 0.05). Our data suggested that pig Bmi1 can increase the proliferation of IPEC-1 cells by promoting the G1/S transition and the overall cell cycle process.

Brain-derived neurotrophic factor (BDNF) plays a critical role in many aspects of neuronal biology and hippocampal physiology and pathology, and has been implicated as a potential therapeutic target in temporal lobe epilepsy (TLE). BDNF total mRNA and its six transcripts were compared in the hippocampal tissue of TLE patients with or without hippocampal sclerosis (HS) by real-time fluorescence quantitative polymerase chain reaction. Excitatory actions induced by BDNF on hippocampal cells were investigated by whole-cell patch-clamp recordings. Statistically significant increases in three human BDNF mRNA transcripts were observed in TLE patients with HS compared with those without HS (transcripts 2, 3 and 5 exhibited 2.1-, 2.3- and 4.1-fold increases, respectively); there were no significant increases in other transcripts. BDNF directly induced N-methyl-D-aspartate currents in dentate granule cells of TLE patients with HS. These results demonstrated that BDNF transcripts were selectively upregulated in TLE patients with HS compared with those without HS. Moreover, BDNF induced excitability of dentate granule cells in TLE patients with HS.

Crossbred pigs were selected for high (HTC) or low (LTC) plasma total cholesterol (TC). Pigs from the seventh (n = 51) and eighth (n = 92) generations were used to determine restriction fragment length polymorphisms (RFLP). Using TaqI restriction enzyme digestion, the frequencies of two alleles (2.8- or 5.0-kb fragments) of the cholesterol 7 alpha-hydroxylase (CYP7) gene were determined in the two populations as a potential indicator of TC concentration at 8 weeks of age. Only the 2.8-kb fragment allele was present in the 26 HTC pigs tested in Generation 7. In the LTC pigs both the 2.8- and 5.0-kb alleles were present in 12 pigs, and only the 5.0-kb allele was present in 13 pigs. The allele frequencies of the 2.8 and 5.0 fragments, respectively, were .26 and .74 in LTC pigs and 1.00 and 0 in HTC pigs. There was an association (P < .001) between the 5.0- and 2.8-kb CYP7 alleles, respectively, and low and high TC concentrations. In Generation 8, all HTC pigs were homozygous for the 2.8-kb allele. The 5.0 kb allele was present in all LTC pigs tested and was homozygous in 57% of LTC pigs. Mean plasma TC was 105.0 mg/dl in 30 pigs homozygous for the 2.8-kb allele in Generation 8; means for LTC pigs were 53.5 and 60.4 mg/dl in 35 pigs homozygous for the 5.0-kb allele and in 27 heterozygous pigs, respectively. High TC was associated with the presence of the 2.8-kb allele, and low TC was associated with the presence of the 5.0-kb allele in both Generations 7 and 8. We conclude that TaqI RFLP analysis of the CYP7 gene is a reliable indicator for TC in these swine.

Background: Skeletal-related events (SREs) occur in 80% of patients with advanced breast cancer (BC) and bone metastases. SREs are costly and can be painful and debilitating, impacting patients’ quality of life and morbidity. While intravenous bisphosphonates such as pamidronate (PAM) and zoledronic acid (ZA) have demonstrated clinical benefit in reducing SREs, skeletal metastases remain a problem as treatments that are more efficacious, well tolerated, more convenient and less costly to administer are needed. Denosumab (XGEVA™) is a novel subcutaneous human monoclonal antibody therapy that significantly reduces the risk of developing SREs in patients with bone metastases from BC. Objective: The objective of this project is to estimate the incremental cost-effectiveness of denosumab relative to ZA and PAM in the treatment of advanced BC patients with bone metastases. Methods: A lifetime Markov model with four-week cycle lengths was developed with three health states: on treatment; off treatment; and death. The model included the risk of an SRE for patients on and off treatment and adverse events during treatment. Efficacy was measured as reduction in SREs. Head-to-head efficacy data, transition probabilities, and risk of adverse events were obtained from the clinical trial of denosumab versus ZA. (Stopeck AT et al JCO 2010) Efficacy data compared to PAM was determined from a published network meta-analysis. (Ford JA et al Eur J Cancer 2012) The baseline SRE risk was derived from clinical trial data due to the absence of real-world Canadian data. Analyses were conducted from the Canadian healthcare system perspective and reported in 2011 $CAD. Resource use was determined from a Canadian retrospective chart review of oncology patients with SREs. Costs were based on the published literature, the Ontario Case Costing Initiative, and input from a physician panel. Utility inputs were based on a time trade-off study. (Matza LS et al. Eur J Health Econ 2013) Bisphosphonate administration costs were derived from a published time and motion study. (Dranitsaris G et al. J Oncol Pharm Pract 2001) Outcomes were measured as both SREs avoided and quality-adjusted life years (QALYs) gained. Dominance was assessed or incremental cost-effectiveness ratios calculated per SRE avoided and per QALY gained, for denosumab compared to ZA and PAM. Future costs and QALYs were discounted at 5% per annum. Sensitivity analyses were conducted to test the robustness of the results. Results: Denosumab was dominant and resulted in $5,733 in cost savings compared to ZA and $2,566 in cost savings compared to PAM based on a probabilistic analysis. Cost savings was driven by differences in drug administration costs and reduction in SREs. SREs avoided were 0.27 and 0.57 compared to ZA and PAM respectively. Denosumab resulted in 0.012 QALYs gained and 0.025 QALYs gained per patient compared to ZA and PAM, respectively. Sensitivity analyses showed the results were robust but most sensitive to drug administration costs and the relative risk of SREs. Conclusion: Compared to both ZA and PAM, denosumab is more efficacious and offers better value for money (i.e. dominant) in Canada for managing SREs in patients with advanced BC and bone metastases. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-07-02.

1.1. Background: Tripterygium Hypoglaucum Hutch (THH), extracts of a Chinese medicinal plant, shows immunosuppressive function and has been used in China for decades to treat autoimmune and inflammatory conditions. This retrospective study aimed to determine long-term efficacy of THH in treating patients with BD-associated uveitis. Methods:Thirty-two patients with BD-associated uveitis resistant to conventional treatment were enrolled. Extracts of THH, 33mg/kg daily, were administered orally along with prednisone. The observation period was 46.8 months. Results:A positive response to THH therapy was observed in 87.5% of patients, showing 62.5% complete remission of uveitis and 75.8% achieved visual acuity maintained or improvement. THH was effective to reduce the incidence of ocular relapse (from 2.9 to 0.4 per yr, p<0.0001) and the dose of corticosteroid (from 22.3 to 11.6 mg daily, p<0.01), with beneficial effects on extraocular manifesta-tions. Side effects are rare and dose-dependent. Conclusion:THH therapy has significant potential in treating patients with sight-threatening BD-associated uveitis, showing beneficial effects on improvement of visual acuity, long-term remission and extraocular lesions of BD.