创伤患者急性肾功能损伤诊断与治疗

目的:探讨患者创伤后发生急性肾损伤的患者发病率、临床特点以及发病危险因素,以便有效预防和及早治疗。方法:回顾性分析我院重症监护室2004年1月至2010年12月收治的创伤患者相关临床资料,分析创伤后急性肾损伤的发病率以及发病危险因素。结果:共有106例患者纳入我们的研究,其中47例患者创伤后并发急性肾损伤。在发生急性肾损伤患者中,平均年龄为31±19岁,84.6%为男性;其中25例为脓毒血症引起,18例是因为低血压导致急性肾功能损伤。所有患者中,24例患者出现了少尿的症状,19例患者进行了透析治疗。腹部外伤[(OR)=3.66,P=0.027]和应用呋塞米[(OR=4.10,P=0.026)]是发生急性肾损伤的危险因素。结论:急性肾损伤时创伤后的严重并发症之一,死亡率高。只有找到创伤后发生急性肾损伤的危险因素,才能有效预防和及早治疗。     

免疫法筛选酵母HSA-IL-11融合蛋白转化子

报道了一种筛选高表达融合蛋白HSA-IL-11的毕赤酵母转化子的免疫双膜筛选法.将生长在醋酸纤维素滤膜上的转化子进行原位诱导,再用硝酸纤维素滤膜对表达的蛋白进行原位捕捉,并经封闭过夜后使用抗HSA抗体进行免疫杂交,再用标记二抗进行显色.根据显色强弱将转化子分为强阳性、中等和阴性三类,再用抗IL-11抗体进行复筛验证.结...     

定向进化提高灰盖鬼伞过氧化物酶染织废水脱色效率

【目的】获得染织废水脱色能力增强的灰盖鬼伞过氧化物酶。【方法】使用基因合成及定点突变平台合成突变灰盖鬼伞过氧化物酶基因CIPmt4(I49S、V53A、M166F和M242I),并调整密码子至毕赤酵母偏好性。以CIPmt4为模板进行定向进化,经过三轮易错PCR和高通量筛选得到一个酶学性质显著改善的突变体(CIPmt5)。通过3D建模和分子动力学模拟分析蛋白的结构及热稳定性,并进一步研究CIPmt5和野生型CIP对刚果红、氨基黑、甲基橙、次甲基蓝、苯胺蓝、结晶紫、溴酚蓝共7种染料的脱色能力。【结果】序列分析显示该突变体积累了I49S、V53A、T121A、M166F和Y272F共5个氨基酸突变,与野生型灰盖鬼伞过氧化物酶相比,以ABTS为底物酶活性是野生型的2.01倍(24.44 U/mg),最适反应p H由5.0提高到6.5,最适反应温度由25°C提高到45°C。除次甲基蓝外对其它染料脱色的最适p H都往中、碱性方向偏移,脱色率普遍高于野生型。模型分析显示CIPmt5活性中心更开放,热稳定性增强。【结论】突变体酶CIPmt5能够更好地替代野生型灰盖鬼伞过氧化物酶应用于染织业染料脱色、化工废水和染织废水的生物修复。     

ABCA1基因多态性R219K与帕金森症和阿尔兹海默症发病率的相关性研究

目的:探讨与研究三磷酸腺苷结合盒转运体A1 (Adenosine triphosphate (ATP)-binding cassette transporter A1)基因多态性R219K与帕金森症(Parkinson disease,PD)和阿尔兹海默症(Alzheimer disease,AD)发病率的相关性。方法:选择2016年2月到2019年8月在本院门诊与住院的帕金森症患者42例作为PD组,同期选择本院门诊与住院的阿尔兹海默症患者42例作为AD组,同期选择本院门诊健康体检者84例作为对照组。调查入选者的一般资料,检测三组血液样本的ABCA1基因多态性R219K情况并进行相关性分析。结果:AD组低密度脂蛋白(low-density lipoprotein,LDL-C)、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)与尿酸(Uric acid,UA)均低于对照组,而高密度脂蛋白(high-density lipoprotein,HDL-C)、同型半胱氨酸(homocysteine,Hcy)值高于对照组(P0.05);AD组TC均低于PD组,而HDL高于PD组。PD患者HDL-C均低于对照组,而LDL、TC和TG与对照组无差异(P0.05),三组空腹血糖(Fasting blood glucose,FBG)值对比差异无统计学意义(P0.05)。PD组与AD组的ABCA1 R219K GA基因型、A等位基因频率都显著高于对照组(P0.05),PD组与AD组对比差异无统计学意义(P0.05)。在168例入选者中,直线相关分析显示ABCA1 R219K GA基因型与A等位基因与帕金森症或阿尔兹海默症发生有显著相关性(P0.05)。结论:ABCA1基因多态性R219K在帕金森症和阿尔兹海默症患者中比较常见,ABCA1 R219K GA基因型与A等位基因可诱发帕金森症和阿尔兹海默症的发生。     

Human alpha-galactosidase A: characterization of the N-linked oligosaccharides on the intracellular and secreted glycoforms overexpressed by Chinese hamster ovary cells

Human alpha-galactosidase A (alpha-Gal A) is the lysosomal glycohydrolase that cleaves the terminal alpha-galactosyl moieties of various glycoconjugates. Overexpression of the enzyme in Chinese hamster ovary (CHO) cells results in high intracellular enzyme accumulation and the selective secretion of active enzyme. Structural analysis of the N -linked oligosaccharides of the intracellular and secreted glycoforms revealed that the secreted enzyme's oligosaccharides were remarkably heterogeneous, having high mannose (63%), complex (30%), and hybrid (5%) structures. The major high mannose oligosaccharides were Man5-7GlcNAc2 species. Approximately 40% of the high mannose and 30% of the hybrid oligosaccharides had phosphate monoester groups. The complex oligosaccharides were mono-, bi- , 2,4-tri-, 2,6-tri- and tetraantennary with or without core-region fucose, many of which had incomplete outer chains. Approximately 30% of the complex oligosaccharides were mono- or disialylated. Sialic acids were mostly N -acetylneuraminic acid and occurred exclusively in alpha2, 3-linkage. In contrast, the intracellular enzyme had only small amounts of complex chains (7.7%) and had predominantly high mannose oligosaccharides (92%), mostly Man5GlcNAc2 and smaller species, of which only 3% were phosphorylated. The complex oligosaccharides were fucosylated and had the same antennary structures as the secreted enzyme. Although most had mature outer chains, none were sialylated. Thus, the overexpression of human alpha-Gal A in CHO cells resulted in different oligosaccharide structures on the secreted and intracellular glycoforms, the highly heterogeneous secreted forms presumably due to the high level expression and impaired glycosylation in the trans- Golgi network, and the predominately Man5-7GlcNAc2 cellular glycoforms resulting from carbohydrate trimming in the lysosome.      

Histological and immunohistochemical studies of the structure of lymph nodes in Kilis goats

Ten healthy adult Kilis goat mesenteric lymph nodes were used to examine the general structure of lymph nodes, lymphocytes, plasma cells, reticular cells and reticular fibers using histological methods. We also detected T lymphocytes using anti-CD3 [SP7], anti-CD4 [74-12-4], mouse anti-bovine CD4 [CC30] and mouse anti-bovine CD8 [CC63] monoclonal antibodies (mAb); and B lymphocytes using anti-CD79a [HM57] mAb, macrophages using anti-macrophage [MAC387] mAb and follicular dendritic cells using anti-S100 polyclonal antibody (pAb). The distribution of these cells also was studied. Although the primer antibodies we used for CD3, CD8, CD79a, MAC387 and S100 worked well, the primer antibodies for CD4 were ineffective for paraffin embedded goat lymph nodes.     

Quantitative trait loci analysis of individual and total isoflavone contents in soybean seeds

Soybean isoflavones play diverse roles in human health, including cancers, osteoporosis, heart disease, menopausal symptoms and pabulums. The objective of this study was to identify the quantitative trait loci (QTL) associated with the isoflavones daidzein (DC), genistein (GeC), glycitein (GlC) and total isoflavone contents (TIC) in soybean seeds. A population of 184 F_21:0 recombinant inbred lines derived from a ‘Xiaoheidou’ בGR8836’ cross was planted in pot and field conditions to evaluate soybean isoflavones. Twenty-one QTL were detected by composite interval mapping. Several QTL were associated with the traits for DC, GeC, GlC and TIC only. QDGeGlTIC4_1 and QDGlTIC12_1 are reported first in this study and were associated with the DC, GeC, GlC and TIC traits simultaneously. The QTL identified have potential value for marker-assisted selection to develop soybean varieties with desirable isoflavone content.