利用等位基因特民扩增法(ASA)为基础的基因分型法,对细胞色素P4502D6 (CYP2D6)缺陷型等位基因携带者的9个家庭共38个进行了基因分型,并与用右旋美沙芬为
探针的表型分型法进行对比,发现两种方法的结果是一致的,CYP2D6酶缺陷型等位基因呈常染色体隐性遗传。
Abstract:A genotyping method based on the principle of allele-specific amplification and a phenotyping procedure with dextromethorphan as a probe were employed in familial study of nine families with 38 members for the cytochrome P450 2D6(CYP2D6)deficient alleles——CYP2D6A,CYP2D6B,CYP2D6D and CYP2D6T.The results showed that the CYP2D6 deficient alleles were inherited as an autosomal recessive trait. 相似文献
Epithelial-mesenchymal transition (EMT) plays an important role in cancer tumorigenesis. However, the underlying mechanisms of EMT in lung adenocarcinoma, and how this process might be inhibited, remain to be explored. This study investigated the role of IL-6 in lung adenocarcinoma cell EMT and explored the potential effects of metformin on this process.
Methods
Invasion assay and MTT assay was performed to determine cell invasion and cell proliferation. Western blotting, immunofluorescence, real-time PCR, ELISA, and immunohistochemistry were performed to detect the expression of IL-6, E-cadherin, Vimentin, and p-STAT3.
Results
We discovered that IL-6, via STAT3 phosphorylation, could promote lung adenocarcinoma cell invasion via EMT in vitro. This was supported by the inverse correlation between E-cadherin and IL-6 expression, positive correlation between IL-6 and vimentin mRNA expression and between STAT3 phosphorylation and IL-6 expression in tumor tissues. Importantly, metformin inhibited tumor growth and distant metastases in tumor-bearing nude mice and reversed IL-6-induced EMT both in vitro and in vivo. Furthermore, we found that blockade of STAT3 phosphorylation might be the underlying mechanism of metformin inhibition of IL-6-induced EMT.
Conclusions
Collectively, our present results show that enhanced IL-6 expression, via STAT3 phosphorylation, is a mechanism of EMT in lung adenocarcinoma. We found that metformin could inhibit IL-6-induced EMT possibly by blocking STAT3 phosphorylation. 相似文献
Olfactory receptors, which are responsible for sensing odor molecules, form the largest G protein-coupled receptor (GPCR) family in mammalian animals. These proteins play an important role in the detection of chemical signals and signal transduction to the brain. Currently, only a limited number of olfactory receptors have been characterized, which is mainly due to the lack of sensitive and efficient tools for performing functional assays of these receptors. This paper describes a novel surface acoustic wave (SAW)-based biosensor for highly sensitive functional assays of olfactory receptors. An olfactory receptor of Caenorhabditis elegans, ODR-10, was expressed on the plasma membrane of human breast cancer MCF-7 cells, which was used as a model system for this study. For specific odorant response assays, the membrane fraction of MCF-7 cells containing ODR-10 was extracted and integrated with our SAW sensors. The response of ODR-10 to various odorants was monitored by recording the resonance frequency shifts of SAWs applied to the sensor. Our results show that heterologously expressed ODR-10 receptors can specifically respond to diacetyl, its natural ligand. Dose-dependent responses were obtained by performing measurements using various concentrations of diacetyl. The sensitivity of this biosensor is 2 kHz/ng and can detect concentrations as low as 10−10 mM, which is 10× lower than what has previously been reported. This biosensor can be used to characterize odorant response profiles of olfactory receptors and provide information rich data for functional assays of olfactory receptors. In addition to providing a greater understanding of the biological mechanisms of GPCRs, such data holds great potential in many other fields such as food industry, biomedicine, and environmental protection. 相似文献
The immobilization efficiency of molecular detectors is of great importance with regard to the performances of biosensors such as the sensitivity, stability, and reproducibility. This paper presents a biomimetic olfactory receptor-based biosensor with better performances by improving the immobilization efficiency of molecular detectors for odorant sensing. A mixed self-assembled monolayers (SAMs) functionalized with specific olfactory receptors (ODR-10) was constructed on the sensitive area of surface acoustic wave (SAW) chip. The immobilization of ODR-10 was characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The responses of this biosensor to various odorants were recorded by monitoring the resonance frequency shifts of SAW, which is correlated to the mass loading on its sensitive area. All the results demonstrate this biosensor can specifically respond to the natural ligand of ODR-10, diacetyl, with high sensitivity and stability. The sensitivity is 4 kHz/ng, which is 2× higher than that of previous work. The detection limit is 1.2×10(-11) mM. The major advances on immobilization efficiency of molecular detectors presented in this work could substantially promote and accelerate the researches and applications of olfactory receptor-based biosensors with different transducers, such as quartz crystal microbalance (QCM), surface plasma resonance (SPR), and field effect transistors (FET). 相似文献
Diabetic neuropathic pain (DNP) is a troublesome diabetes complication all over the world. P2X3 receptor (P2X3R), a purinergic receptor from dorsal root ganglion (DRG), has important roles in neuropathic pain pathology and nociceptive sensations. Here, we investigated the involvement of DRG P2X3R and the effect of 2 Hz electroacupuncture (EA) on DNP. We monitored the rats’ body weight, fasting blood glucose level, paw withdrawal thresholds, and paw withdrawal latency, and evaluated P2X3R expression in DRG. We found that P2X3R expression is upregulated on DNP, while 2 Hz EA is analgesic against DNP and suppresses P2X3R expression in DRG. To evaluate P2X3R involvement in pain modulation, we then treated the animals with A317491, a P2X3R specific antagonist, or α β-me ATP, a P2X3R agonist. We found that A317491 alleviates hyperalgesia, while α β-me ATP blocks EA’s analgesic effects. Our findings indicated that 2 Hz EA alleviates DNP, possibly by suppressing P2X3R upregulation in DRG.
Oligochitosan, which has greater than 3 but less than 10 saccharide (N-acetylglucosamine or glucosamine) residues, is obtained by either chemical or enzymatic hydrolysis of chitosan. In this work, we demonstrated that oligochitosan had an in vitro stimulatory effect on the release of tumor necrosis factor-alpha and interleukin-1 beta in macrophages. Moreover, we observed that oligochitosan could be uptaken by macrophages through confocal laser microscopy. Scatchard analysis of internalization of 2-aminoacridone-oligochitosan in macrophages indicated its internalization was mediated by a specific receptor on macrophage membrane with a Kd of 2.1 x 10(-5) M. Competition studies showed that mannose could inhibit oligochitosan internalization, while lipopolysaccharide and beta-glucan could not do it. Inhibition of mannose-BSA, fucose-BSA, and N-acetylglucosamine-BSA on oligochitosan internalization further suggests that oligochitosan internalization is mediated by a macrophage lectin receptor like with mannose specificity. 相似文献