人Leptin基因的克隆及其在大肠杆菌中的表达

目的克隆Leptin基因,构建其重组表达菌株,并对表达产物进行免疫学鉴定。方法利用RT-PCR方法从脂肪细胞RNA中扩增出人瘦素前体(pre-Leptin)的cDNA片段和去信号肽序列的Leptin成熟基因片段,并插入pET32a中,转化大肠杆菌BL21(DE3),获得表达重组人瘦素(rh-Leptin)的菌株。结果DNA序列分析显示获得的pre-Leptin和成熟基因片段的序列与预计序列一致,菌株诱导后经SDS-PAGE检测,rh-Leptin表达量达菌体总蛋白的40%以上。Western blot分析显示重组Lep-tin具有免疫学活性。结论已获得高效表达Leptin的重组菌株。     

LPS诱导的胸腺细胞凋亡对瘦素的抑制作用

要提高声表面波(SAW)气体传感器的测量精确度,温度补偿是主要难题.目前有许多补偿方法,但其效果不佳.采用软件方法进行温度补偿的研究在国内外已成热点,但选用神经网络对声表面波气体传感器进行温度补偿罕见报道.该文以西北工业大学研发的声表面波CO气体传感器为研究对象,通过理论分析和实验,得到了声表面波CO气体传感器的温度特性曲线.提出了一种利用BP人工神经网络对声表面波CO气体传感器温度误差进行修正的新方法.计算机仿真和试验结果表明,该法能有效改善传感器的输出特性,且速度快,精度高,鲁棒性强,便于用硬件实现,具有较高的推广应用价值.     

Dietary Supplementation of Calcium may Counteract Obesity in Mice Mediated by Changes in Plasma Fatty Acids

The scope of this study was to assess the impact of calcium and conjugated linoleic acid (CLA) supplementation on plasma fatty acid profiles and to evaluate potential synergistic effects of both compounds against dietary obesity. Mice separated into five experimental groups were followed: control (C), high-fat diet (HF), HF with calcium (Ca), HF plus CLA and HF with both Ca and CLA. Plasma metabolites and fatty acids were determined by commercial kits and gas chromatography, respectively. Both dietary calcium and CLA supplementation contributed to lower body fat gain under a HF diet. Maximum efficacy was seen with calcium; no additional effect was associated with the combined treatment with CLA. Plasma leptin, adiponectin and HOMA index were in accordance with an altered glucose/insulin homeostasis in the HF and HF + CLA groups, whereas control levels were attained under Ca-enriched diets. Plasma fatty acids showed minor changes associated to CLA treatment, but a high impact on PUFA was observed under Ca-enriched diets. Our results show that the mechanism underlying the anti-obesity effects of calcium supplementation is mediated mainly by changes in PUFA plasma profile. In addition, the lack of synergy on body weight reduction in combination with associated lipid profiles of calcium and CLA suggests that calcium may interfere with absorption and/or bioactivity of CLA, which can be of relevance when using CLA-fortified dairy products against human obesity.     

Leptin promotes proliferation and invasion of osteosarcoma cells by upregulating the expression of SIRT1

Osteosarcoma (OS) is a primary high-grade malignant bone neoplasm, and the prognosis of OS remains poor due to early metastasis. Leptin plays an essential role in tumorigenesis, but the role of leptin in the development of OS is still not fully understood. In this study, we used a human osteosarcoma MG-63 cell line as an experimental model. MG-63 cells were treated with leptin, and cell proliferation, apoptosis, adhesion, invasion, and gene expression, were evaluated. The results showed that leptin promoted proliferation, decreased adhesion, suppressed apoptosis, and promoted invasion, of MG-63 cells. Moreover, the expression of SIRT1 was upregulated in MG-63 cells exposed to leptin. Furthermore, MMP-2, 8, and 9 were significantly upregulated by SIRT1, while SIRT1 knockdown inhibited the proliferation and invasion of MG-63 cells. In conclusion, our results suggest that leptin promotes OS cell proliferation and invasion by inducing the expression of SIRT1.     

Effect of the genistein metabolite on leptin secretion in murine adipocytes in vitro

Soy isoflavonoids have many useful properties. However, they are metabolized in vivo, including in humans. The effect of the metabolism of soy isoflavonoids on their properties is not fully understood. We have isolated the bacterial strain SY8519, which has been shown to metabolize daidzein to O-desmethylangolensin and to produce 2-(4-hydroxyphenyl)propionic acid from genistein. According to chiral HPLC analysis, the 2-(4-hydroxyphenyl)propionic acid obtained from the bacterium was optically active. To determine the absolute stereochemistry of the microbial product, we prepared (S)-2-(4-hydroxyphenyl)propionic acid from (S)-2-phenylpropionic and concluded that the microbial product had an R-configuration by chiral HPLC analysis. We also applied the metabolite to mouse adipocytes and found that 2-HPPA was less effective at reducing leptin secretion than the parent compound genistein. Our results suggested that ‘O-desmethylangolensin-production’ attenuates the effect of soy isoflavonoids by reducing not only the activity of daidzein but also that of genistein.     

Fatty Acid-Specific Alterations in Leptin,PPARα, and CPT-1 Gene Expression in the Rainbow Trout

It is known that fatty acids (FA) regulate lipid metabolism by modulating the expression of numerous genes. In order to gain a better understanding of the effect of individual FA on lipid metabolism related genes in rainbow trout (Oncorhynchus mykiss), an in vitro time‐course study was implemented where twelve individual FA (butyric 4:0; caprylic 8:0; palmitic (PAM) 16:0; stearic (STA) 18:0; palmitoleic16:1n‐7; oleic 18:1n‐9; 11‐cis‐eicosenoic 20:1n‐9; linoleic (LNA) 18:2n‐6; α‐linolenic (ALA) 18:3n‐3; eicosapentenoic (EPA) 20:5n‐3; docosahexaenoic (DHA) 22:6n‐3; arachidonic (ARA) 20:4n‐6) were incubated in rainbow trout liver slices. The effect of FA administration over time was evaluated on the expression of leptin, PPARα and CPT‐1 (lipid oxidative related genes). Leptin mRNA expression was down regulated by saturated fatty acids (SFA) and LNA, and was up regulated by monounsaturated fatty acids (MUFA) and long chain PUFA, whilst STA and ALA had no effect. PPARα and CPT‐1mRNA expression were up regulated by SFA, MUFA, ALA, ARA and DHA; and down regulated by LNA and EPA. These results suggest that there are individual and specific FA induced modifications of leptin, PPARα and CPT‐1 gene expression in rainbow trout, and it is envisaged that such results may provide highly valuable information for future practical applications in fish nutrition.     

Is there any interaction of resistin and adiponectin levels with protein‐energy wasting among patients with chronic kidney disease

The aim of this study was to evaluate the effects of adipocytokines including adiponectin, leptin, resistin, neuropeptide Y and ghrelin in chronic kidney disease (CKD) patients on appearance of protein‐energy wasting (PEW). One hundred fifty patients with mean age of 45.4 ± 15.9 years, without active infections or chronic inflammatory conditions were recruited into the study. Study groups were control group (consisting of 30 healthy volunteers with normal kidney functions), hemodialysis group, predialysis group, peritoneal dialysis group and kidney transplant group. Fasting morning serum leptin, ghrelin, acylated ghrelin, neuropeptide Y, adiponectin, resistin levels of all of the groups were measured. Anthropometric and nutritional assessments of all patients were obtained. Diagnosis of PEW was made according to definition recommended by the International Society of Renal Nutrition and Metabolism. Presence of PEW in hemodialysis (23.3%) and peritoneal dialysis (26.7%) groups were significantly higher than those of predialysis (3.3%), and transplantation (0%) groups. Adiponectin and resistin levels in predialysis, peritoneal dialysis and hemodialysis patients were significantly higher than control group (p: 0.0001). This study had given significant positive correlations between presence of PEW and serum resistin (r: 0.267, p: 0.001), and serum adiponectin levels (r: 0.349, p: 0.0001). There were no relationship between presence of PEW and ghrelin, acylated‐ghrelin, neuropeptide Y, and leptin levels of the groups. CKD patients except transplant patients had higher adiponectin and resistin levels than control group. PEW was found to be linearly correlated with resistin and adiponectin. High serum resistin and adiponectin levels might have a role in development of PEW among dialysis patients.