Protein microarrays enhanced performance using nanobeads arraying and polymer coating

A nanosize material composed of 330 nm glass beads coated with a copolymer of N,N-dimethylacrylamide (DMA), N,N-acryloyloxysuccinimide (NAS) and [3-(methacryloyl-oxy)propyl]trimethoxysilane (MAPS) was developed to improve the protein immobilization on biochips. The developed material, bearing rabbit-IgG proteins, was arrayed as 150 μm spots trapped at the surface of a poly(dimethylsiloxane) elastomer (PDMS), and compared to copoly(DMA-NAS-MAPS)-coated glass slides and latex beads based biochips. Evidences were made through scanning electron microscopy that the newly developed material based microarray exhibited surface irregularities at the submicron level leading to high specific area.The combination of such large immobilization area with the highly efficient protein immobilization of the copoly(DMA-NAS-MAPS) polymer, enabled the achievement of microarrays exhibiting good performances both in pure media and complex samples (human sera). Indeed, high specific/non-specific signal ratio was found using this optimized immobilization procedure.Chemiluminescent detection of anti-rabbit-IgG was obtained through peroxidase labeled antibodies in the 5 μg/l to 10 mg/l range. Application of the developed system to real samples was achieved for the detection of rheumatoid factor (RF) through a capture assay. Interesting results were obtained, with a RF detection over the 5.3-485 IU/ml range and without measurable matrix effect or non-specific signal.     

Nanotechnology for diagnosis and therapy of rheumatoid arthritis: Evolution towards theranostic approaches

Rheumatoid arthritis(RA),as a chronic autoimmune disease,damages the bone and cartilage of patients,and even leads to disability.Therefore,the diagnosis and treatment of RA is particularly important.However,due to the complexity of RA,it is difficult to make effective early diagnosis of RA,which is detrimental to RA treatment.Besides,long-term intake of anti-RA drugs can also cause damage to patients' organs.The emergence of nanotechnology provides the new train of thoughts for the diagnosis and treatment of RA.And the combination of diagnosis and therapy is an ideal method to solve the problem of disease management of RA patients.In this review,we summarize the mechanism and microenvironment of RA,discuss the commonly used diagnostic techniques and therapeutic drugs for RA,and review their advantages and disadvantages.New nanotherapy strategies such as drug-carrying nanoparticles,PTT,PDT are listed,and their applications in RA treatment are also summarized.In addition,multimodal imaging,combined therapy and responsive diagnosis and treatment are also summarized as important contents.At last,we also review typical nanocarriers that can be used in the integration of diagnosis and therapy,and discussed their potential applications in RA theranostics.     

Selective chemiluminescence method for monitoring of vitamin K homologues in rheumatoid arthritis patients

Vitamin K is a fat-soluble vitamin involved in blood coagulation and bone metabolism. The detection and monitoring of vitamin K homologues in rheumatoid arthritis (RA) patients is a challenging problem due to the smaller concentrations of vitamin K and the presence of several interfering medications. Therefore, this study aimed to develop a new highly sensitive and selective chemiluminescence (CL) method designated to quantify vitamin K homologues in plasma of RA patients including phylloquinone (PK, vitamin K₁), menaquinone-4 (MK-4, vitamin K₂) and menaquinone-7 (MK-7, vitamin K₂). The method was based on the unique photochemical properties of vitamin K homologues that were exploited for selective luminol CL reaction. The correlation coefficients of 0.998 or more were obtained in the concentration ranges of 0.1-100 ng mL⁻¹ vitamin K homologues. The detection limits were 0.03-0.1 ng mL⁻¹ in human plasma for vitamin K homologues. The developed HPLC-CL system was successfully applied for selective determination of vitamin K homologues in plasma of RA patients. The developed method may provide a useful tool for monitoring vitamin K homologues in different clinical studies such as RA, osteoporosis and hepatocellular carcinoma in which vitamin K is intervented.     

类风湿关节炎患者血浆置换治疗前后微量元素铜的变化

应用原子吸收分光光度法，对１５例类风湿关节炎（ＲＡ）患者血浆置换前后微量元素铜的变化进行了测定。结果表明，血浆置换后患者的血铜水平显著下降，提示血浆置换治疗ＲＡ存在铜元素的机制。     

Latex piezoelectric immunoassay: Effect of interfacial properties

Latex piezoelectric immunoassay is a technique for detecting agglutination of antibody- or antigen-bearing latex by an immunoreaction using a piezoelectric quartz crystal; the agglutination decreases the oscillation frequency of the crystal. This is advantageous in that immobilization of antibody or antigen on the crystal surface is unnecessary. In this report, different kinds of chemical functional groups were immobilized on the electrode surface, allowing us to consider the effect of interfacial structure on the frequency change. Electrode modifications such as self-assembly of alkanethiol and aminoalkoxysilane monolayers, and polyethylenimine-glutaraldehyde coating as well as plasma treatment were examined. The sensitivity of the system was found to imitate the interfacial properties so that modification of the electrode surface could improve the response. Among the methods examined for this electrode surface modification, the polyethylenimine-glutaraldehyde modification had the advantages of high reproducibility, fast operation and simplicity. It was also suggested that the frequency change originated primarily from the immunoreaction at the interface.     

Liquid chromatographic method for the simultaneous determination of sulphasalazine and its main metabolites in biological fluids

Summary An HPLC procedure was developed for the simultaneous determination of salicylazosulphapyridine, and its main metabolites 5-aminosalicylic acid and sulphapyridine, in human serum and synovial fluid. The analytical procedure consisted of a single ion-pair extraction step for an Extrelut column with methylene chloride. The investigated compounds and the added sulphadimidine internal standard were eluated from a Hypersil-MOS reversed-phase column by stepwise gradient; mobile phase was methanol-0.01 M potassium dihydrogenphosphate (3:7, 0.0–2.0 min and 8:2, 2.1–6.5 min).     

邻苯二胺-过氧化氢-辣根过氧化物酶酶联免疫示差脉冲伏安法测定人血清类风湿因子抗体

建立了邻苯二胺（ＯＰＤ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）酶联示差脉冲伏安分析体系并用于测定人血清中类风湿因子（ＲＦ），ＨＲＰ催化Ｈ２Ｏ２氧化ＯＰＤ所形成酶催化产物在ｐＨ２．０磷酸盐－枸橼酸缓冲溶液中于－０．１８ Ｖ左右产生一灵敏示差脉冲伏安峰，ＲＦ浓度在１．２５～２０．０ Ｕ／ｍＬ之间与峰电流呈线性关系，应用此峰检测人血清ＲＦ的检测限低至 ０．２８Ｕ／ｍＬ。该法较相同条件下ＥＬＩＳＡ显色光度测定法的灵敏度增加５倍，且受干扰较少。     

对211例类风湿关节炎患者微量元素的测定

用同位素激发Ｘ射线荧光分析对２１１例类风湿关节炎患者发中１０种常量及微量元素测定，以探索类风湿关节病与微量元素之间的关系．结果表明：全部女性患者发锌含量明显降低，除３０～３９岁及５０～５９岁年龄组Ｐ＜０．０５外，其余各组均Ｐ＜０．０１。而２０～２９岁男性组患者发锌含量也低，Ｐ＜０．０１。男女患者各组发锶含量较对照组均低，且有明显差异（除男性２０～２９岁组及５０￣５９岁组Ｐ＜０．０５，其余各组均ｐ＜０．０１）。提示类风湿关节病患者发中锌、锶含量较正常人低。因此，如临床上补充锌和锶元素，可能是有效治疗类风湿关节病的新途径．     

Label-free detection of rheumatoid factor using YbYxOy electrolyte–insulator–semiconductor devices

In this study, we investigated the effect of yttrium content on the structural properties and sensing characteristics of YbY_xO_y sensing membranes for electrolyte–insulator–semiconductor (EIS) sensors to detect the rheumatoid factor (RF). The YbY_xO_y EIS device prepared at the 60 W plasma condition exhibited a higher sensitivity of 65.77 mV/pH, a lower hysteresis voltage of ∼1 mV, and a smaller drift rate of 0.14 mV/h than did those prepared at the other conditions. We attribute this behavior to the optimal yttrium content in the YbY_xO_y film forming a smooth surface. Furthermore, we used a novel YbTi_xO_y EIS biosensor to measure the RF antigen in human serum because of its rapid and label-free detection. Two different techniques were used for the immobilization of RF antibody onto the surface of an YbTi_xO_y EIS sensor. The RF antibody was directly immobilized on the EIS surface modified with 3-aminopropyltriethoxysilane (APTES) followed by glutaraldehyde (GA). In contrast, a mixture of 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) solution was used to functionalize the carboxyl groups at the tail of RF antibodies. RF antibodies functionalized with the active NHS esters were covalently immobilized on the APTES-modified YbTi_xO_y surface. The immobilized RF antibodies on the EIS that are functionalized with the EDC and NHS exhibit higher (41.11 mV/pC_RF) for detection of serum RF antigen in the range 10⁻⁷ to 10⁻³ M, compared to traditional antibody immobilization technique via APTES and GA linkage. The YbTi_xO_y EIS biosensor is a promising analytical tool for RF antigen monitoring due to its good sensitivity, stability and repeatability.     

计算机模式识别法在类风湿中医证侯的研究

根据计算机模式识别的马氏(Mahalanobis)距离判别法，用BASIC等语言缩制的IBM-PC等徽机适用的多元统计判别程序将健康人分别与类风湿性关节炎的寒热错杂型、寒湿阻络型及肝肾两虚型的发样徽量元素Zn、Fe、Cu、Cd及Zn／Cu五种因素进行分类判别．结果表明以Zn、Ft、Zn／Cu为特征参量．健康人与寒热情杂型类风温性关节炎患者分类准确亭为95．6%，而分别以Zn、Cu和Zn，Fe、Cu为特征参量．健康人与寒湿阻络型和肝肾两虚型患者分类准确率分别为100％和90．2％．说明以Fe、Cu、Zn(或Zn／Cu)组成的微量元素谱含量，在不同中医证型的类风温性关节炎和健康中是有差异的。