荧光光谱研究CBAP与人血清白蛋白的相互作用

在模拟人体生理条件下,用荧光光谱和紫外可见吸收光谱法研究了不同温度下4-(2-羧基苯偶氮)-连苯三酚(CBAP)与人血清白蛋白(HSA)的相互作用.结果表明,CBAP对HSA的荧光猝灭属静态猝灭.通过288、293、298 K时的荧光猝灭,得出CBAP与HSA的结合常数K分别为1.62×106,3.97×105和3.56×105,结合位点数n=0.92(平均值).据F(o)rster非辐射能量转移理论计算出CBAP与HSA的结合距离r=1.44 nm,由热力学参数焓变和熵变,推断两者反应时氢键和范德华力可能起主要作用.运用同步荧光和三维荧光技术分析了CBAP对HSA构象的影响,表明CBAP的加入改变了HSA中酪氨酸残基和色氨酸残基的构象.     

氢氯噻嗪与牛血清白蛋白的相互作用

用荧光光谱法研究了氢氯噻嗪(Hydrochlorothiazide,HCT)与牛血清白蛋白(BSA)的相互作用。实验结果表明,HCT与BSA作用的猝灭常数随着温度的升高而降低,HCT可以有规律地使BSA内源荧光猝灭,其猝灭机理可认为是HCT与BSA形成复合物的静态猝灭。通过测定和计算不同温度下该结合反应的结合常数和结合位点数,并根据热力学方程求得了结合反应的ΔG、ΔH和ΔS等热力学参数,根据所得结果推断出HCT与BSA间的主要作用力类型是疏水作用力。同时,从分子荧光寿命进一步证明HCT与BSA的荧光猝灭机制为静态猝灭。     

牛血清白蛋白和苯甲酸钠相互作用的荧光光谱研究

利用荧光光谱研究了牛血清白蛋白(BSA)与苯甲酸钠(SB)的相互作用,结果表明,随着苯甲酸钠浓度的增加,BSA荧光强度逐渐减弱。二者以1:1的结合比结合,考察了不同pH、离子强度、不同金属离子,表面活性剂的条件下,苯甲酸钠与BSA的结合情况。     

八羧酸酞菁钴配合物与牛血清白蛋白的结合作用研究

八羧基金属酞菁的周边四个苯环各带有两个羧基,具有较好的水溶性,且吸收峰在670-700nm,是一类性能优良的光敏剂,而且CoPc(COOH)8带有负电荷,对蛋白有很强的亲和作用。采用光谱法研究了CoPc(COOH)8与牛血清白蛋白(BSA)的结合作用;采用荧光光谱分析方法测定了CoPc(COOH)8与BSA的结合常数为K=6.94×105;以氯血红素(HE)、布洛芬(IB)、L-色氨酸(TRP)为BSA的分子配体,络合竞争法研究了它们对CoPc(COOH)8-BSA体系紫外光谱的影响,并且确定了CoPc(COOH)8与BSA的结合点为SiteIII。     

Cu（Ⅱ）与牛血清白蛋白相互作用的荧光光谱研究

利用荧光光谱技术,从分子水平上研究了模拟生理条件下Cu2＋与牛血清白蛋白（BSA）的相互作用机理。实验结果表明,Cu2＋使得内部疏水区域的色氨酸（Trp）等芳香环氨基酸逐步暴露出来,在荧光光谱中表现为BSA内源荧光的猝灭,猝灭机理主要为静态猝灭。Cu2＋与BSA的结合常数为8.73×104,结合比为1：1。     

地美硝唑与牛血清白蛋白结合热力学特征的荧光光谱研究

采用荧光光谱和紫外可见吸收光谱法研究了地美硝唑与牛血清蛋白的结合反应,发现地美硝唑对牛血清白蛋白有较强的荧光猝灭作用,该猝灭过程主要为静态猝灭过程。从荧光光谱数据,由Stern—Volmer方程和Lineweaver—Burk方程分析并处理实验数据得到了23℃时结合反应的结合常数K=1．03×10^4L／mol,结合位点数为0．70,结合反应的标准焓变、标准熵变、和标准吉布斯自由能变分别为～63．48kJ／mol,-96．22J／K,-35．00kJ／mol。     

近生理浓度的脲对核黄素与牛血清蛋白结合作用的影响

用CD光谱和荧光光谱研究了近生理浓度的脲存在和不存在两种情况下核黄素和牛血清蛋白之间的相互作用.一定浓度的核黄素结合在牛血清蛋白上可以引起BSA分子结构伸展,二级结构和三级结构减少.近生理浓度的脲可以诱导核黄素变性的牛血清蛋白二级结构和三级结构部分恢复.脲存在时,核黄素对牛血清蛋白的熄灭常数（Kq）以及供体和受体之间的距离（r）增大.相反地,结合常数（Ka）和结合位点数（n）减小.脲存在下,核黄素和牛血清蛋白的结合作用是一个熄灭作用增强,结合作用减弱的过程.人体内血脲的正常范围为0.0025–0.0075M,脲浓度达到0.01 M时,由于核黄素和HAS分子的弱结合作用导致血浆中核黄素缺乏,一些病人可能会因为核黄素缺乏,结合着药物的生物大分子不能被有效地转移进细胞内,不能被某些药物有效地治疗.     

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC₅₀) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%–84.60%, with coefficients of variation of less than 12.2%.     

N-十六烷基-羟乙基-二甲基溴化铵与牛血清白蛋白的结合作用

用荧光光谱法研究了298K时Tris-HC l缓冲溶液（pH=7.1）中季铵盐类表面活性剂N-十六烷基-羟乙基-二甲溴化铵（CHDAB）与牛血清白蛋白（BSA）的结合作用.考察了BSA浓度对结合作用的影响,用Stern-Volmer方程探讨了CHDAB在浓度较低区域与BSA的作用机制,用位点结合模型计算了CHDAB与BSA结合反应的结合常数KA和结合位点数n,用同步荧光技术考察了CHDAB对BSA构象的影响.结果表明：CHDAB对BSA的内源荧光有猝灭作用,并导致其最大发射波长蓝移;相同温度下,BSA浓度越小,其Stern-Volmer猝灭常数Ksv越大,CHDAB对BSA的猝灭作用越强,同时其结合常数也越大,结合越强;同步荧光光谱表明CHDAB对BSA构象产生了一定影响.     

荧光光谱法研究Fe~（2＋）和Fe~（3＋）与牛血清白蛋白的竞争结合作用

利用荧光光谱法、紫外吸收光谱法、同步荧光法和三维荧光法研究了金属离子Fe2＋、Fe3＋与牛血清白蛋白（BSA）在模拟生理条件下的竞争结合作用.结果表明,Fe2＋、Fe3＋对BSA的内源荧光都有明显的猝灭作用,猝灭机理均为静态猝灭和动态猝灭的联合作用,主导作用分别为静态猝灭和动态猝灭,并分别计算了结合常数、结合位点数和热力学参数,Fe3＋与BSA的结合能力强于Fe2＋.弱的结合力使BSA与Fe2＋、Fe3＋相互作用后构型没有明显改变.当Fe2＋、Fe3＋同时与BSA相互作用时,二者表现出弱的竞争取代现象.     

荧光光谱法研究甲磺酸酚妥拉明与血红蛋白的结合作用

药物摄入人体之后,与血液中的血红蛋白结合,从而使血红蛋白的结构和功能发生改变.因此,研究甲磺酸酚妥拉明与血红蛋白的相互作用机理有助于阐明药物在生物体内的运输和代谢机制、药物的毒理机制,以及了解蛋白质的结构与功能关系都具有非常重要的意义.本文用荧光光谱技术研究了甲磺酸酚妥拉明与牛血红蛋白的相互作用机理,确定了相互作用的结合常数及结合位点数.     

Anti-senescence effect and molecular mechanism of the major royal jelly proteins on human embryonic lung fibroblast (HFL-I) cell line

Royal jelly (RJ) from honeybee has been widely used as a health promotion supplement. The major royal jelly proteins (MRJPs) have been identified as the functional component of RJ. However, the question of whether MRJPs have anti-senescence activity for human cells remains. Human embryonic lung fibroblast (HFL-I) cells were cultured in media containing no MRJPs (A), MRJPs at 0.1 mg/ml (B), 0.2 mg/ml (C), or 0.3 mg/ml (D), or bovine serum albumin (BSA) at 0.2 mg/ml (E). The mean population doubling levels of cells in media B, C, D, and E were increased by 12.4%, 31.2%, 24.0%, and 10.4%, respectively, compared with that in medium A. The cells in medium C also exhibited the highest relative proliferation activity, the lowest senescence, and the longest telomeres. Moreover, MRJPs up-regulated the expression of superoxide dismutase-1 (SOD1) and down-regulated the expression of mammalian target of rapamycin (MTOR), catenin beta like-1 (CTNNB1), and tumor protein p53 (TP53). Raman spectra analysis showed that there were two unique bands related to DNA synthesis materials, amide carbonyl group vibrations and aromatic hydrogens. These results suggest that MRJPs possess anti-senescence activity for the HFL-I cell line, and provide new knowledge illustrating the molecular mechanism of MRJPs as anti-senescence factors.     

荧光法研究司他夫定与牛血清白蛋白的结合作用

在Tris缓冲溶液（pH7．1）体系中,用荧光光谱技术研究水溶液中司他夫定与牛血清白蛋白的结合作用．结果表明,司他夫定对牛血清白蛋白内源荧光产生较强的荧光猝灭作用,根据不同温度下司他夫定对牛血清白蛋白的荧光猝灭作用,证明其为静态猝灭机制,运用位点模型计算出298K,310K时其结合常数KA（分别为1．01×10^4,4．17×10^4L·mol^-1）和结合位点数n（分别为0．99,0．93）,根据热力学参数确定其作用力以氢键或VanderWall＇s作用为主;蛋白质变性剂尿素的存在导致上述荧光猝灭效率减少;运用FSster偶极-偶极非辐射能量转移原理,计算了司他夫定与牛血清白蛋白的结合距离r为3．79nm．     

荧光光谱法研究芥子碱与人血清蛋白的相互作用

在模拟生理条件下(pH=7.4),采用荧光光谱法研究了芥子碱与人血清蛋白(HSA)的相互作用。结果表明芥子碱可以使人血清蛋白的内源荧光发生猝灭。根据Stern-Volmer方程计算得到不同温度下的荧光猝灭常数,在296～303 K温度范围为静态猝灭,303~310 K为动态猝灭;由Lineweaver-Burk双倒数方程得出不同温度下芥子碱和HSA的结合常数(Ka)和结合位点数(n),并计算得到其荧光猝灭的吉布斯自由能(G)和熵值(S),其结果表明芥子碱与人血清蛋白的作用过程是一个熵增加、自由能降低的自发反应。在296~303 K时两者凭借氢键和范德华力进行结合,在303~310 K温度范围两者的主作用力为疏水作用。     

荧光光谱法研究黄豆苷原与牛血清白蛋白的相互作用

用荧光光谱法在pH7.4的PBS中研究了黄豆苷原（Daidzein,Da）与牛血清白蛋白（bovine serum albu-min,BSA）的相互作用.结果表明Da可静态猝灭BSA的内源荧光,温度升高时受动态猝灭的影响变得明显.二者之间的结合位点数约为1.5,315 K、310 K,300 K和290 K时的表观结合常数分别为5.44×104L.mol-1,5.70×104L.mol-1,5.29×104L.mol-1和5.69×104L.mol-1.焓变为0.2969 kJ.mol-1,熵变为91.78为J.mol-1.K-1,各温度下的自由能变均小于零,热力学参数表明二者之间主要靠静电力发生作用,熵变对反应过程具有较大贡献,且反应是自发过程.同步荧光光谱表明Da的加入改变了BSA的构像,使色氨酸残基的微环境亲水性增强.本研究对于阐明Da与蛋白的相互作用机理具有借鉴意义.     

Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice

Human serum albumin(HSA) is widely utilized for medical purposes and biochemical research.Transgenic rice has proved to be an attractive bioreactor for mass production of recombinant HSA(rHSA).However,transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical value.This study aimed to develop a selectively terminable transgenic rice line expressing HSA in rice seeds,and a simple process for recovery and purification of rHSA for economical manufacture.An HSA expression cassette was inserted into a T-DNA vector encoding an RNA interference(RNAi) cassette suppressing the CYP81A6 gene.This gene detoxifies the herbicide bentazon and is linked to the 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) cassette which confers glyphosate tolerance.ANX Sepharose Fast Flow(ANX FF) anion exchange chromatography coupled with Butyl Sepharose High Performance(Butyl HP) hydrophobic interaction chromatography was used to purify rHSA.A transgenic rice line,HSA-84,was obtained with stable expression of rHSA of up to 0.72% of the total dry weight of the dehusked rice seeds.This line also demonstrated high sensitivity to bentazon,and thus could be killed selectively by a spray of bentazon.A two-step chromatography purification scheme was established to purify the rHSA from rice seeds to a purity of 99% with a recovery of 62.4%.Results from mass spectrometry and N-terminus sequencing suggested that the purified rHSA was identical to natural plasma-derived HSA.This study provides an alternative strategy for large-scale production of HSA with a built-in transgene safety control mechanism.     

Influence of serum adiponectin level and SNP +45 polymorphism of adiponectin gene on myocardial fibrosis

Adiponectin plays an important role in the development of hypertension, atherosclerosis, and cardiomyocyte hypertrophy, but very little was known about the influence of serum adiponectin or the adiponectin gene polymorphism on myocardial fibrosis. Our study investigates the influence of the SNP +45 polymorphism of the adiponectin gene and serum levels of adiponectin on myocardial fibrosis in patients with essential hypertension. A case-control study was conducted on 165 hypertensive patients and 126 normotensive healthy controls. The genotypes of adiponectin gene polymorphisms were detected by the polymerase chain reaction (PCR) method. Serum concentrations of procollagen were measured by a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) in all subjects. The integrated backscatter score (IBS) was measured in the left ventricular myocardium using echocardiography. The serum levels of adiponectin in hypertensive patients were significantly lower than those in the normal control group ((2.69±1.0) μg/ml vs. (4.21±2.89) μg/ml, respectively, P<0.001). The serum levels of type-I procollagen carboxyl end peptide (PICP) and type-III procollagen ammonia cardinal extremity peptide (PIIINP) in the hypertension group were significantly higher than those in the control group. In the hypertension group, serum levels of adiponectin were significantly and negatively related to the average acoustic intensity and corrected acoustic intensity of the myocardium (r=0.46 and 0.61, respectively, P<0.05 for both). The serum levels of PICP and PIIINP were significantly different among the three genotypes of SNP +45 (P<0.01). Logistic regression analyses showed that sex and genotype (GG+GT) were the major risk factors of myocardial fibrosis in hypertensive patients (OR=5.343 and 3.278, respectively, P<0.05). These data suggest that lower levels of adiponectin and SNP +45 polymorphism of the adiponectin gene are likely to play an important role in myocardial fibrosis in hypertensive patients.     

腐殖酸和牛血清蛋白对氧化物纳米颗粒团聚和沉降行为的影响

研究目的：比较牛血清蛋白和腐殖酸对Al2O3、SiO2和TiO2三种纳米颗粒团聚与沉降行为的影响,并讨论其影响机制。创新要点：纳米颗粒团聚物直径的增大能引起其沉降速度的加快,但小的水动力学直径并不一定导致低的沉降速度,说明团聚直径不是决定纳米颗粒沉降的唯一因素。研究方法：通过透射电镜观察纳米颗粒团聚物的形态;采用动态光散射技术研究纳米颗粒的团聚动力学;最后通过测量悬浊液的光学吸收来研究纳米颗粒的沉降动力学。重要结论：牛血清蛋白处理降低了三种纳米颗粒在NaCl和CaCl2中的水动力学直径,原因是牛血清蛋白的球状结构能够引起颗粒间的空间位阻斥力。腐殖酸处理导致纳米颗粒的水动力学直径在NaCl中最小,而在CaCl2中最大（图4）,原因是腐殖酸能通过钙的配位作用彼此连接,从而促进了纳米颗粒的团聚。牛血清蛋白减缓了纳米颗粒的沉降;然而腐殖酸在CaCl2中明显加大了纳米颗粒的沉降速度（图5）,与其水动力学直径的增大一致。腐殖酸处理的纳米颗粒在NaCl中水动力学直径最小,但沉降速度却不是最低。     

匹伐他汀钙对高胆固醇血症患者动脉粥样硬化的影响（英文）

目的:观察匹伐他汀钙对高胆固醇血症患者外周血管的影响。创新点:首次在国内发现匹伐他汀钙能够改善高胆固醇血症患者肱动脉和颈动脉血管内皮功能而且延缓其动脉粥样硬化发展,并首次证实改善内皮功能是匹伐他汀钙延缓其动脉粥样硬化发展的重要原因。方法:按照入选排除标准,选取本院高胆固醇血症患者(HC),完成超声心动图检查的40例。根据剂量不同,分为两个剂量组:1 mg剂量组20例(男性5例,女性15例,平均年龄(55.20±8.35)岁),2 mg剂量组20例(男性9例,女性11例,平均年龄(57.56±6.09)岁)。访视结束后完成超声心动图检查的HC组36例,两个剂量组分别有2人失访。治疗后1 mg剂量组18例(男性3例,女性15例,平均年龄(56.00±7.85)岁),2 mg剂量组18例(男性7例,女性11例,平均年龄(57.79±6.46)岁)。选择本院同期体检中心30例正常人作为对照(年龄和性别均与病例组匹配,男性14例,女性16例,平均年龄(54.94±6.90)岁)。所有研究对象,均经隔夜禁食12~14小时,次日清晨抽取空腹肘静脉血,测定临床生化指标。采用Sequia512彩色多普勒超声诊断仪,应用高分辩率外周血管超声技术,检测HC治疗前后肱动脉血流介导性舒张功能(FMD)、颈动脉结构和功能。结论:经匹伐他汀钙治疗8周后,高胆固醇血症患者血管功能明显改善,表现为FMD升高,僵硬度减小;颈动脉僵硬度和内中膜厚度(IMT)延缓进展与其内皮功能改善密切相关。     

Charlson合并症指数对2型糖尿病肾病死亡率预测作用的研究(英文)简

Our intent is to examine the predictive role of Charlson comorbidity index （CCI） on mortality of patients with type 2 diabetic nephropathy （DN）. Based on the CCI score, the severity of comorbidity was categorized into three grades： mild, with CCI scores of 1-2; moderate, with CCI scores of 3-4; and severe, with CCI scores 〉5. Factors influencing mortality and differences between groups stratified by CCI were determined by logistical regression analysis and one-way analysis of variance （ANOVA）. The impact of CCI on mortality was assessed by the Kaplan- Meier analysis. A total of 533 patients with type 2 DN were enrolled in this study, all of them had comorbidity （CCI score 〉1）, and 44.7% （238/533） died. The mortality increased with CCI scores： 21.0% （50/238） patients with CCI scores of 1-2, 56.7% （135/238） patients with CCI scores of 3-4, and 22.3% （53/238） patients with CCI scores 〉5. Logistical regression analysis showed that CCI scores, hemoglobin, and serum albumin were the potential predictors of mortality （P〈0.05）. One-way ANOVA analysis showed that DN patients with higher CCI scores had lower levels of hemoglobulin, higher levels of serum creatinine, and higher mortality rates than those with lower CCI scores. The Kaplan-Meier curves showed that survival time decreased when the CCI scores and mortality rates went up. In con- clusion, CCI provides a simple, readily applicable, and valid method for classifying comorbidities and predicting the mortality of type 2 DN. An increased awareness of the potential comorbidities in type 2 DN patients may provide insights into this complicated disease and improve the outcomes by identifying and treating patients earlier and more effectively.