摘要：
Although draft genome sequences of two of the major human schistosomes, Schistosoma japonicum and Schistosoma mansoni are available, the structures and characteristics of most genes and the influence of exogenous genes on the metabolism of schistosomes remain uncharacterized. Furthermore, which functional genomics approaches will be tractable for schistosomes are not yet apparent. Here, the vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter, under the control of the retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce S. japonicum to investigate the utility of retrovirus-mediated transgenesis of S japonicum and the activity of human telomerase reverse transcriptase as a reporter transgene in schistosomes. Schistosomules perfused from experimentally infected rabbits were cultured for 6 days after exposure to the virions after which genomic DNAs from virus exposed and control worms were extracted. Analysis of RNA from transduced parasites and immunohistochemistry of thin parasite sections revealed expression of hTERT in the transduced worms. Expression of hTERT was also confirmed by immunoblot analysis. These findings indicated that S. japonicum could be effectively transduced by VSVG-pseudotyped retrovirus carrying the hTERT gene. Given the potential of hTERT to aid in derivation of immortalized cells, these findings suggest that this pantropic retroviral approach can be employed to transduce cells from specific tissues and organs of schistosomes to investigate the influence of transgene hTERT on growth and proliferation of schistosome cells. (C) 2010 Elsevier B.V. All rights reserved.
作者：
Yan Liu;Qingren Zeng;Jun Zhou;Shenghui Yang;Biyuan Liu;...
作者机构：
Department of Parasitology,School of Medicine,University of South China;Center of cell and Molecula;Center of cell and Molecular Biology Experiment,Xiangya School of Medicine,Central South University;Experimental Center of Xiangya Third Hospital,CSU;Department of Pathogenic Biology and Immunology,Hunan University of Chinese Medicine
作者：
Yang, S. H.;Brindley, P. J.;Zeng, Q. R.*;Zeng, T. B.;Li, Y. S.;...
作者机构：
[Li, Y. S.; Liu, Y.; Cai, L. T.; Liu, B. Y.; Zeng, T. B.; Lan, L. M.; Yang, S. H.; Zeng, Q. R.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.;[Wu, C. R.; Tan, Z. J.; Lu, F. G.; Yang, S. H.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.;[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.;[Liu, Y.; Liu, B. Y.; Zeng, T. B.] Univ South China, Sch Med, Hengyang, Peoples R China.;[Li, Y. S.; McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.
会议名称：
12th International Congress of Parasitology (ICOPA)/6th Novel Approaches to the Control of Helminth Parasites of Liverstock Conference
会议时间：
AUG 15-20, 2010
会议地点：
Melbourne, AUSTRALIA
会议主办单位：
[Yang, S. H.;Zeng, Q. R.;Zeng, T. B.;Li, Y. S.;Liu, Y.;Liu, B. Y.;Cai, L. T.;Lan, L. M.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.^[Yang, S. H.;Wu, C. R.;Lu, F. G.;Tan, Z. J.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.^[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.^[Zeng, T. B.;Liu, Y.;Liu, B. Y.] Univ South China, Sch Med, Hengyang, Peoples R China.^[Li, Y. S.;McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.^[Zhou, J.] Cent S Univ, Expt Ctr Xiangya Third Hosp, Changsha, Hunan, Peoples R China.
摘要：
Retroviral transduction of cultured schistosomes was explored in our study. The vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter gene under the control of retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce Schistosoma japonicum (Sj) to investigate the utility of retrovirus mediated Sj transgenesis and to investigate the activity as human telomerase as a reporter transgene in schistosomes. The presence of transgene hTERT was detected by PCR. Furthermore, analysis of RNAs from transduced parasites, immunohistochemistry of thin sections and immunoblot analysis revealed expression of hTERT transgene in the transduced worms. These findings indicated that Sj could be effectively transduced by VSVG pseudotyped retrovirus carrying the hTERT gene.
通讯机构：
[Yin, Weidong] U;Univ S China, Life Sci Res Ctr, Inst Cardiovasc Res, Key Lab Atherosclerol Hunan Prov, Hengyang 421001, Hunan, Peoples R China.
摘要：
Insulin resistance and dyslipidemia are both considered to be risk factors for metabolic syndrome. Low levels of IGF1 are associated with insulin resistance. Elevation of low-density lipoprotein cholesterol (LDL-C) concomitant with depression of high-density lipoprotein cholesterol (HDL-C) increase the risk of obesity and type 2 diabetes mellitus (T2DM). Liver secretes IGF1 and catabolizes cholesterol regulated by the rate-limiting enzyme of bile acid synthesis from cholesterol 7 alpha-hydroxylase (CYP7A1). NO-1886, a chemically synthesized lipoprotein lipase activator, suppresses diet-induced insulin resistance with the improvement of HDL-C. The goal of the present study is to evaluate whether NO-1886 upregulates IGF1 and CYP7A1 to benefit glucose and cholesterol metabolism. By using human hepatoma cell lines (HepG2 cells) as an in vitro model, we found that NO-1886 promoted IGF1 secretion and CYP7A1 expression through the activation of signal transducer and activator of transcription 5 (STAT5). Pretreatment of cells with AG 490, the inhibitor of STAT pathway, completely abolished NO1886-induced IGF1 secretion and CYP7A1 expression. Studies performed in Chinese Bama minipigs pointed out an augmentation of plasma IGF1 elicited by a single dose administration of NO-1886. Long-term supplementation with NO-1886 recovered hyperinsulinemia and low plasma levels of IGF1 suppressed LDL-C and facilitated reverse cholesterol transport by decreasing hepatic cholesterol accumulation through increasing CYP7A1 expression in high-fat/high-sucrose/high-cholesterol diet minipigs. These findings indicate that NO-1886 upregulates IGF1 secretion and CYP7A1 expression to improve insulin resistance and hepatic cholesterol accumulation, which may represent an alternative therapeutic avenue of NO-1886 for T2DM and metabolic syndrome. Journal of Endocrinology (2010) 204, 47-56
摘要：
To develop a minipig model of type 2 diabetes that simulates the common manifestations of the metabolic abnormalities and resembles the kidney pathology of type 2 diabetes in the human population, male Chinese Bama minipigs were divided into 2 groups (5 in each) and fed with a control diet (CD) or high-fat/ high-sucrose/ high-cholesterol diet (HFSCD) for 5 months. The biochemical parameters of blood and urine, and the oral glucose tolerance test were monitored after the feeding program. The insulin resistance was estimated by the HOMA-IR index and the glucose elimination constant (K(G)), and beta-cell function by the HOMA-beta index and the acute insulin response (AIR). Glomerulosclerosis index (GSI) was semi-quantitated by the degree of glomerular lesions in kidney sections stained with Masson trichrome. Extracellular matrix deposition in the kidney was examined by the protein expression of type IV collagen, connective tissue growth factor (CTGF) and matrix metalloproteinases 2 (MMP-2) using immunohistochemistry. Feeding HFSCD to minipigs markedly caused hyperglycaemia, hyperinsulinaemia and dyslipidaemia. HOMA-IR was significantly increased while HOMA-beta, AIR and K(G) were obviously decreased in the HFSCD group compared with control group. Microalbuminuria, glucosuria and moderate glomerulosclerosis were exhibited in HFSCD-fed minipigs. The expression of type IV collagen and CTGF was elevated whereas that of MMP-2 was reduced in the kidneys of HFSCD group compared with the CD group. We concluded that feeding HFSCD to Chinese Bama minipigs for 5 months can induce humanoid type 2 diabetes and early-stage diabetic nephropathy, and accelerate extracellular matrix deposition and glomerulosclerosis.
摘要：
Inflammation, closely associated with obesity, is emerging as an important risk factor for the pathophysiological development of atherosclerosis and diabetes mellitus. Fat balance is critical in the aetiology of obesity. Lipoprotein lipase is an important enzyme in lipid metabolism. The aim of this study was to investigate the long-term effect of the lipoprotein lipase activator, NO-1886, on inflammation cytokines, adiposity and related diseases in miniature pigs fed a high-fat/high-sucrose/high-cholesterol diet (HFSC diet). Chinese Bama-miniature pigs were fed a control diet or HFSC diet with or without NO-1886 for 5 months. The levels of inflammation-associated cytokines were determined using the antibody arrays. Feeding of the HFSC diet to miniature pigs markedly increased the expression of inflammatory cytokines. On the other hand, supplementation of NO-1886 to HFSC diet decreased the expression of inflammatory cytokines significantly, protecting against the development of atherosclerosis and diabetes mellitus. NO-1886 may have a beneficial effect on the most inflammation-associated cytokines, and this effect may contribute to improving atherosclerosis and diabetes mellitus.
