解析COMT基因Met多态与子宫内膜异位症的关系

目的探讨COMT基因Met多态与子宫内膜异位症的相关性。方法该研究选择的观察组对象60例,均为该院收治的子宫内膜异位症者,与40例对照组非内异症及腺肌病的女性采用PCR-RFLP法进行COMT基因分型,回顾相关资料。结果观察组野生纯和型(G/G)基因型、突变杂合型(G/A)基因型、突变纯合型基因型频率与对照组比较差异无统计学意义(P>0.05)。两组G等位基因频率(观察组72.9%,对照组27.1%)与A等位基因频率(观察组70%,对照组30%)比较差异无统计学意义(P>0.05)。COMTC/G(OR=1)、G/A(OR=0.516,95%CI为0.186-1.442)、A/A(OR=0.451,95%CI=0.166-1.234),3种基因型相对危度分析差异无统计学意义(P>0.05)。提示子宫内膜异位症的发生与COMT基因多态无明显相关性。结论子宫内膜异位症的发生与COMT基因Met多态无相关性,在临床治疗中可作为参考依据。     

人源抗Met基因工程抗体scFv的改造与特性分析

目的:将已制备的抗Met单链抗体基因与人IgG Fc基因片段融合,表达有活性的融合蛋白分子,以增强单链抗体的可溶性.方法:从人淋巴细胞提取总RNA,经RT-PCR扩增并制备人IgG的Fc基因片段,并克隆于已构建好的pBAD-scFv原核表达载体中,转化大肠杆菌Top10,经阿拉伯糖诱导表达融合蛋白scFv-Fc.所表达的可溶性蛋白经亲和层析纯化、SDS-PAGE、Western blot分析鉴定,并用ELISA检测抗体效价.结果:序列分析表明重组质粒pBAD-scFv-Fc基因序列正确;SDS-PAGE分析表明,scFv-Fc融合蛋白分子量为60 ku,且为可溶性蛋白;该蛋白经过His亲和层析纯化、ELISA检测,结果表明,该融合蛋白能够与抗原分子Met特异性结合.结论:改造后的抗体融合蛋白scFv-Fc能与人Met特异性结合,增加了抗体蛋白溶解度,有利于抗体的大量制备.     

Immunocytochemical survey of haloperidolinduced immunoreactive changes of [Met]enkephalin-Arg-Gly-Leu in the rat forebrain

It has already been demonstrated that chronic treatment with the dopamine receptor blocker, haloperidol, results in an increase of proenkephalin-A-derived peptides in the caudate-putamen (CP). To examine this phenomenon at the cellular level, we used immunocytochemistry to investigate the effects of haloperidol on [Met]enkephalin-Arg⁶-Gly⁷-Leu⁸ (MEAGL) immunoreactivity in the rat forebrain. After daily haloperidol (5 mg/kg, IP, for 6 days) or haloperidol decanoate (70 mg/kg, IM, given once or twice) treatment, immunoreactive neurons appeared diffusely in the whole CP and in the core part of the nucleus accumbens (Acb) and less frequently in the outer shell part of the Acb and the cell-dense layer of the tuberculum olfactorium (TuO). Increase of MEAGL-immunoreactive fibers in the CP, Acb and TuO was also detected after these treatments, a particularly prominent increase being found in the striopallidal terminals in the globus pallidus and ventral pallidum. Haloperidol or haloperidol decanoate had no effect on MEAGL immunoreactivity in the cerebral cortex, amygdala, or hypothalamus. Reserpine treatment (5 mg/kg, IP, for 6 days) caused similar effects on the dorsal and ventral striopallidal system and the direct injection of 6-hydroxydopamine (10 μ/5 μl) into the CP led to the appearance of MEAGL-immunoreactive neurons in accordance with the depleted dopaminergic terminal area. These findings suggest that haloperidol influences enkephalinergic neurons region specifically and that in the dorsal and ventral striopallidal enkephalinergic system haloperidol increases MEAGL immunoreactivity in cell bodies, fibers and terminals by blocking intrastriatal dopaminergic neurotransmission.     

Corneal transplantation in a patient with mucopolysaccharidosis type VII (Sly disease)

Purpose: To illustrate a good visual outcome following penetrating keratoplasty in a patient with Sly disease, a rare mucopolysaccharidosis (MPS) caused by a deficiency of ß-glucuronidase. Methods: A 15-year-old male with progressive bilateral corneal opacification had a complete medical, genetic, and ophthalmic evaluation followed by a penetrating keratoplasty. Results: The cornea has remained clear for two years following surgery. Histopathology of the corneal button demonstrated vacuoles and granular inclusions consistent with this lysosomal storage disease. Conclusion: While research is ongoing in the fields of enzyme replacement and bone marrow transplantation, these treatments may not alleviate or reverse the corneal clouding. This case illustrates that cornea transplantation may be a valuable treatment option for visually rehabilitating such patients.     

Gender differences in methionine accumulation and metabolism in freshly isolated mouse hepatocytes: Potential roles in toxicity

l-Methionine (Met) is hepatotoxic at high concentrations. Because Met toxicity in freshly isolated mouse hepatocytes is gender-dependent, the goal of this study was to assess the roles of Met accumulation and metabolism in the increased sensitivity of male hepatocytes to Met toxicity compared with female hepatocytes. Male hepatocytes incubated with Met (30 mM) at 37 °C exhibited higher levels of intracellular Met at 0.5, 1.0, and 1.5 h, respectively, compared to female hepatocytes. Conversely, female hepatocytes had higher levels of S-adenosyl-l-methionine compared to male hepatocytes. Female hepatocytes also exhibited higher l-methionine-l-sulfoxide levels relative to control hepatocytes, whereas the increases in l-methionine-d-sulfoxide (Met-d-O) levels were similar in hepatocytes of both genders. Addition of aminooxyacetic acid (AOAA), an inhibitor of Met transamination, significantly increased Met levels at 1.5 h and increased Met-d-O levels at 1.0 and 1.5 h only in Met-exposed male hepatocytes. No gender differences in cytosolic Met transamination activity by glutamine transaminase K were detected. However, female mouse liver cytosol exhibited higher methionine-dl-sulfoxide (MetO) reductase activity than male mouse liver cytosol at low (0.25 and 0.5 mM) MetO concentrations. Collectively, these results suggest that increased cellular Met accumulation, decreased Met transmethylation, and increased Met and MetO transamination in male mouse hepatocytes may be contributing to the higher sensitivity of the male mouse hepatocytes to Met toxicity in comparison with female mouse hepatocytes.     

Effect of functional catechol-O-methyltransferase Val158Met polymorphism on physical aggression

Genetic and psychological analysis of the relationships between catechol-O-methyltransferase Val158Met polymorphism and various types of aggressiveness was performed in 114 women. Dispersion analysis revealed significant association of ValVal genotype with elevated physical aggression. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 1, pp. 68–70, January, 2008     

Knockdown of c-Met enhances sensitivity to bortezomib in human multiple myeloma U266 cells via inhibiting Akt/mTOR activity

The c-Met is a receptor tyrosine kinase that is overexpressed in human myeloma cell lines and promotes the survival and drug resistance of myeloma cells. This study aimed to elucidate the mechanisms by which c-Met contributes to the chemoresistance in myeloma. Stable U266 cell line in which c-Met was effectively knockdown was employed and treated with bortezomib. Cytotoxicity was evaluated by MTT assay. Cell cycle profile and apoptosis were examined by cytometry analysis. The expression of cell cycle related proteins, and the activities of caspases and Akt/mTOR were detected by Western blot analysis. The c-Met knockdown in U266 cells decreased the average IC(50) of bortezomib, induced G0/G1 phase arrest, and increased caspase-mediated apoptosis in U266 cells exposed to bortezomib. In addition, c-Met knockdown decreased the level of cyclin D1 and increased the levels of p27 and cleaved caspase 3 and caspase 9. Moreover, the Akt/mTOR activity in U266 cells treated with bortezomib was downregulated upon c-Met knockdown and c-Met knockdown U266 cells recovered chemoresistance upon the overexpression of Akt and mTOR. Our data demonstrate that c-Met is a potential therapeutic target for multiple myeloma, and Akt/mTOR is a key signaling component through which c-Met protects multiple myeloma cells from chemotherapy-induced growth inhibition and apoptosis.     

Iron, ferritin and proteins of the methionine-centered redox cycle in young and old rat hearts

Progressive oxidation of cellular components constitutes a major mechanism of the aging process. An emerging paradigm of redox signaling suggests that low level oxidants activate protective pathways resulting in prolonged cell survival. This report centers on the study of cardiac muscle in young and old rats, including (i) the expression of ferritin (Ft) the major iron storage protein, and (ii) the expression of the major proteins of the methionine-centered redox cycle (MCRC), which controls the cellular methionine redox status. Total amounts of Ft (protein) and its mRNA encoding for Ft L-subunit (Ft-L) were higher in the aged hearts, indicating that the iron-binding capacity of myocardial Ft increased with age. Among the proteins of the MCRC, methionine sulfoxide reductases A and B (MsrA, MsrB) and MsrA mRNA were significantly higher in hearts of old rats with a significant decrease in MsrA activity. The observed up-regulation of the expression of Msr and Ft-L could represent a protective response to the increased oxidative stress in the aging myocardium.     

Brain‐derived neurotrophic factor Val66Met polymorphism and early life adversity affect hippocampal volume

The interaction between adverse life events during childhood and genetic factors is associated with a higher risk to develop major depressive disorder (MDD). One of the polymorphisms found to be associated with MDD is the Val66MET polymorphism of brain‐derived neurotrophic factor (BDNF). The aim of our two‐center study was to determine how the BDNF Val66Met polymorphism and childhood adversity affect the volumetric measures of the hippocampus in healthy individuals and people with MDD. In this two‐center study, 62 adult patients with MDD and 71 healthy matched controls underwent high‐resolution magnetic resonance imaging. We used manual tracing of the bilateral hippocampal structure with help of the software BRAINS2, assessed childhood adversity using the Childhood Trauma Questionnaire and genotyped Val66Met BDNF SNP (rs6265). MDD patients had smaller hippocampal volumes, both in the left and right hemispheres (F = 5.4, P = 0.022). We also found a significant interaction between BDNF allele and history of childhood adversity (F = 6.1, P = 0.015): Met allele carriers in our samples showed significantly smaller hippocampal volumes when they did have a history of childhood adversity, both in patients and controls. Our results highlight how relevant stress–gene interactions are for hippocampal volume reductions. Subjects exposed to early life adversity developed smaller hippocampal volumes when they carry the Met‐allele of the BDNF polymorphism. © 2013 Wiley Periodicals, Inc.