Summary Steady-state expression quantitative trait loci (eQTLs) explain only a fraction of disease-associated loci identified through genome-wide association studies (GWAS), while eQTLs involved in gene-by-environment (GxE) interactions have rarely been characterized in humans due to experimental challenges. Using a baboon model, we found hundreds of eQTLs that emerge in adipose, liver, and muscle after prolonged exposure to high dietary fat and cholesterol. Diet-responsive eQTLs exhibit genomic localization and genic features that are distinct from steady-state eQTLs. Furthermore, the human orthologs associated with diet-responsive eQTLs are enriched for GWAS genes associated with human metabolic traits, suggesting that context-responsive eQTLs with more complex regulatory effects are likely to explain GWAS hits that do not seem to overlap with standard eQTLs. Our results highlight the complexity of genetic regulatory effects and the potential of eQTLs with disease-relevant GxE interactions in enhancing the understanding of GWAS signals for human complex disease using nonhuman primate models.
OBJECTIVE To investigate the effects of heme oxygenase-1 (HO-1) gene on human islets in vitro, and to explore the potential value of gene therapy in clinical islet transplantation. METHODS Adenovirus vector carrying human HO-1 gene (Ad-HO-1) or EGPF (Ad-EGFP) were established respectively. Human cadaveric pancreases were isolated, purified, cultured, and divided into 3 groups to be transfected with Ad-HO-1, Ad-EGFP or blank vector. Human tumor necrosis factor and cyclohexamide (CHX) were added into the culture fluid of the pancreatic islets. 48 hours later the pancreatic islets were digested into single cells. Flow cytometry was used to detect the apoptosis. Glucose of the concentration of 16.7 mmol/L was added into the culture fluid of the 3 groups of islet cells. After 1-hour co-incubation radioimmunochemistry was used to detect the level of insulin in the supernatant. RESULTS After stimulation of glucose the insulin concentration in the supernatant of the Ad-HO-1 group was 270 mIU/L +/- 89 mIU/L, significantly higher than those of the Ad-EGFP group (189 mIU/L +/- 88 mIU/L) and control group (182 mIU/L +/- 59 mIU/L, both P < 0.05). The apoptotic ratio of the Ad-HO-1 group was 63.1% +/- 10.9%, significantly lower than that of the control group (90.9% +/- 11.3%, P < 0.01) after treatment with TNFalpha and CHX. CONCLUSION Transfection of Ad-HO-1 into human islets improves anti-apoptotic function in cultured human islets and promotes insulin release of human pancreatic islets.
Abstract Background The mechanism of erectile dysfunction (ED) caused by low androgen status is not fully understood. Objectives To investigate whether low androgen status inhibits erectile function of rats by inducing pyroptosis in the corpus cavernosum (CC). Materials and methods Thirty‐six eight‐weeks‐old healthy male Sprague‐Dawley rats were equally divided into six groups: sham‐operated group (4w sham, 8w sham), castration group (4w cast, 8w cast), and castration + testosterone (T) group (4w cast + T, 8w cast + T). The rats in castration + T groups were injected with testosterone propionate subcutaneously every other day. After 4 and 8 weeks, the ratio of maximum intracavernous pressure (ICPmax)/mean arterial pressure (MAP), the level of serum T, the concentration of nitric oxide (NO) and interleukin‐1β (IL‐1β), the expression of NOD‐like receptor pyrin domain containing 3 (NLRP3), apoptosis‐associated speck‐like protein containing a caspase activation and recruitment domain (ASC), Caspase‐1 p20, gasdermin D‐N (GSDMD‐N), transforming growth factor β1 (TGF‐β1), collagen‐I, and collagen‐III, the ratio of smooth muscle/collagen (SM/C), and the proportion of pyroptotic cells in the CC were analyzed. Results The ratio of ICPmax/MAP (3/5 V) and SM/C, the level of NO and serum T was significantly decreased in castration groups when compared to other groups ( p < 0.01). NLRP3, ASC, Caspase‐1, and GSDMD were mainly expressed in the cytoplasm of smooth muscle cells (SMCs) and endothelial cells (ECs) in the CC. The expression of NLRP3, ASC, Caspase‐1p20, GSDMD‐N, IL‐1β, TGF‐β1, collagen‐I, and collagen‐III was significantly increased in castration groups when compared with other groups ( p < 0.01). The proportion of pyroptotic cells in the CC was increased significantly in castration groups when compared with other groups ( p < 0.05). Discussion and Conclusion Low androgen status inhibits erectile function of rats by promoting CC fibrosis and reducing NO synthesis through pyroptosis of SMCs and ECs in the CC.
Objective To investigate the efficiency of ileal transposition on bodyweight,gut hormones glucagon-like peptide-1 (GLP-1) and glucose tolerance of Yorkshire piglets.Methods 17 Yorkshire young pigs were randomized into IT group and Sham group which accepted ileal transposition and Sham operation respectively.The bodyweight was recorded weekly.Duodenum glucose tolerance tests wer performed in all pigs on POW4.Consecutive blood samples were collected for glucose measurement,the area under glycemic curves were calculated.Blood samples of T0 min and T40 min were selected for GLP-1 examination.Results No bodyweight difference is identified between IT and Sham group in POW4 (Z =0.42,P ＞ 0.05).In POW4,the peak value of glycemia in IT group (10.58 ± 2.85) mmol/L is significant lower than that of preoperative baseline (13.5 ± 2.56) mmol/L,(Z =2.31,P ＜ 0.05),and the glycemic peak-time is significantly delayed in IT group (90 ± 10.54) min when compared with Sham group (50.00 ± 5.16) min,(Z =2.29,P ＜ 0.05) ; Compared with the 180 min AUC of baseline (1938.4 ± 873.4) mmol/L × min,the IT group (1568.9 ± 546.3) mmol/L × min is significant lower (Z =2.26,P ＜0.05) during the DGTT;40 min after glucose administration,the GLP-1 level of IT group (10.0 ± 1.56 ng/L) is significantly higher than in Sham group (4.27 ± 1.68 ng/L,Z =2.12,P ＜ 0.05).Conclusion Ileal transposition could effectively enhance the level of the gut hormone GLP-1,lower the peak value of blood glucose,defer the peak time and improve the glucose tolerance of Yorkshire piglets in early operative stage,and the effects are independent with weight loss.
Key words:
Ileal transposition; Glucagon-like peptide-l ; Porcine models; Glucose tolerance
Objective: To explore the effects of nuclear M-CSF on the process of tumorigenesis. Methods: Functional part of M-CSF cDNA was inserted into an eukaryotic expression plasmid pCMV/myc/nuc, which can add three NLS to the C-terminal of the expressed protein and direct the protein into the cell nuclei. The constructed plasmid was transferred into NIH3T3 cells and the cell clones were selected by G-418 selection. Cell clones stable expressing target protein were identified by RT-PCR, ABC immunohistochemistry assay and Western blot. Cell growth kinetics analyses through growth curves, cell doubling time, MTT test and anti-sense oligodeoxynucleotide (ASODN) inhibiting cell growth test were performed to identify cells proliferation potential. Results: The transfected cells showed elevated proliferation potential over the control cells. Conclusion: Abnormal appearance of M-CSF in nucleus could enhance cell proliferation, which suggests that cytokine isoforms within cell nucleus might play transcription factor-like role.
HIV-1 protease (PR) is an essential enzyme for viral production. Thus, PR inhibitors (PIs) are the most effective class of anti-HIV drugs. However, the main challenge to the successful use of PI drugs in patient treatment is the emergence of multidrug resistant PRs (mdrPRs). This study aimed to develop a fission yeast cell-based system for rapid testing of new PIs that combat mdrPRs. Three mdrPRs were isolated from HIV-infected patients that carried seven (M7PR), ten (M10PR) and eleven (M11PR) PR gene mutations, respectively. They were cloned and expressed in fission yeast under an inducible promoter to allow the measurement of PR-specific proteolysis and drug resistance. The results showed that all three mdrPRs maintained their abilities to proteolyze HIV viral substrates (MA↓CA and p6) and to confer drug resistance. Production of these proteins in the fission yeast caused cell growth inhibition, oxidative stress and altered mitochondrial morphologies that led to cell death. Five investigational PIs were used to test the utility of the established yeast system with an FDA-approved PI drug Darunavir (DRV) as control. All six compounds suppressed the wildtype PR (wtPR) and the M7PR-mediated activities. However, none of them were able to suppress the M10PR or the M11PR. The three clinically isolated mdrPRs maintained their viral proteolytic activities and drug resistance in the fission yeast. Furthermore, those viral mdrPR activities were coupled with the induction of growth inhibition and cell death, which could be used to test the PI activities. Indeed, the five investigational PIs and DRV suppressed the wtPR in fission yeast as they did in mammalian cells. Significantly, two of the high level mdrPRs (M10PR and M11PR) were resistant to all of the existing PI drugs including DRV. This observation underscores the importance of continued searching for new PIs against mdrPRs.
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