阿尔茨海默病生物标志物及其在新药研发中的应用

理想的生物标志物是实现阿尔茨海默病（AD）早期精确诊断和预测疾病发展的重要前提。新版AD 诊断标准从临床无症状期到 痴呆阶段的区分也有赖于生物标志物的应用。介绍目前临床应用的AD 生物标志物及其在药物研发中的应用,重点对AD 血液生物标志 物研究的现状和瓶颈进行综述。     

尿液作为新型生物标志物来源的探索

机体各种变化是生物标志物研究的核心内容.因为机体固有的稳态调控机制,在血液中早期产生的变化容易被清除.而在尿液中不存在稳态调控机制,允许容纳更多的变化因素存在.尤其在疾病发生的早期阶段,从尿液中更有可能发现新型的早期生物标志物.在尿液生物标志物研究中,亦应当考虑药物的影响.使用尿液生物标志物研究路线图,能够有效规避各种影响因素对于尿液生物标志物研究的干扰;同时,结合膜处理新技术,有利于经济、高效地大规模收集尿液样本,促进尿液生物标志物的大规模研究.另外,本文介绍了最容易在尿液中体现出变化的肾脏疾病生物标志物的研究进展.尿液生物标志物的研究,将赋予人类在疾病预防、诊断、治疗及预后监测等诸多方面实现更多进步的可能性.     

冠心病相关循环miRNA

冠心病诊断中最主要的挑战就是从常规的血液样本中鉴定出可靠的临床生物标志物.循环miRNA是一种可以稳定存在于体液中的小分子RNA,具有较高的组织、疾病特异性及敏感性,具有作为新的冠心病非侵入性生物标志物的潜力.通过综述血液样本（全血、血浆、血清、外周血单核细胞（PBMC）、内皮祖细胞（EPC）及血小板）中冠心病相关循环miRNA,及探讨循环miRNA研究中存在的一些问题,为未来筛选出真正具有临床应用价值的循环miRNA生物标志物奠定基础.     

临床蛋白质组学———蛋白质组学在临床研究中的应用

临床蛋白质组学是将蛋白质组学技术应用于临床医学研究,它主要围绕疾病的预防、早期诊断和治疗等方面开展研究,其中,恶性肿瘤是临床蛋白质组学研究的一个重点研究对象.由于肿瘤生物标志物对早期诊断具有重要价值,所以临床蛋白质组学的主要目标之一是寻找合适的肿瘤生物标志物,多分子生物标志物已成为寻找肿瘤生物标志物的一个研究趋势.简要介绍了临床蛋白质组学的基本概念,实验设计,临床样本收集与预处理以及蛋白质组学技术在临床研究中的应用与进展.     

综述与专论：阿尔茨海默病体外诊断纳米技术

阿尔茨海默病(Alzheimer’s disease,AD)是一种最常见的神经退行性疾病。AD的精准诊断技术,特别是早期诊断技术是临床亟需的。近年来,以生物标志物为基础的非侵入性体外诊断技术发展迅速,特别是利用纳米材料和纳米技术的高表面活性、独特的光电特性、生物相容性好、易于表面修饰、小型化、集成化等特点,发展了新型的AD体外检测和诊断的纳米技术,大大提升了AD检测的灵敏度和准确性,并且具有简便快速等特点,在AD疾病的早期诊断、预后判断以及疗效评估等方面发挥着重要的作用。本文综述了AD蛋白质类生物标志物检测纳米技术的研究进展,介绍了纳米材料在生物标志物富集方面发挥的重要作用,阐述了以纳米材料为基础的光电信号转导技术以及增强检测信号和提高检测灵敏度的方法。除此之外,还简要介绍了AD纳米检测技术在临床诊断、预后和疗效评估方面的应用前景,总结了AD体外诊断技术的优势及面临的挑战,为AD精准诊疗研究提供参考信息。     

骨关节炎相关生物标志物

骨关节炎（osteoarthritis）是关节软骨进展的退化性疾病,并累及周围组织结构的病变,是致老年人伤残主要原因之一。目前,以临床表现和影像学诊断为主,缺乏早期检测和预后评估的有效方法。生物标志物的检查是具有前景的研究方向,在关节软骨结构改变之前,各种生物标志物代谢发生变化,其能帮助诊断和预测骨关节炎的发生发展及其预后。然而,生物标志物在临床诊断和治疗相关的应用仍需加以证实。通过广泛查阅近年有关骨关节炎相关分子生物诊断的相关文献,有助于了解生物标志物对于骨关节炎的早期诊断意义和临床应用前景。本文就关节软骨、骨和滑膜等不同组织类型相关的生物标志物进行综述。     

生物标志物在前列腺炎中的研究进展

前列腺炎是男性泌尿系统的常见疾病,由于前列腺炎存在病因和发病机制复杂,导致临床症状表现多种、药物治疗缺乏特异性和复发率较高等问题,因此前列腺炎的诊断、治疗和预后判断标准一直是临床关注的焦点。目前还没有诊断前列腺炎的"金标准",而生物标志物的检测有助于临床诊断、治疗和预后诊断,本文检索近年相关,对敏感性较高和特异性较好的一些前列腺炎生物标志物(细胞因子和趋化因子等)及今后研究的侧重点进行综述,从而为前列腺炎的诊断、分型治疗和预后观察提供依据。     

非靶标~1H-NMR尿液代谢组学筛选生物标志物组用于心理亚健康的诊断识别

心理亚健康(psychological suboptimal health)临床症状表现多样,且无与其密切相关的具体生化指标,因而心理亚健康的判别和诊断成为目前研究中的重点和难点。本研究旨在发现并评价能够用于临床诊断的尿液生物标志物组,为心理亚健康的临床诊断提供依据。通过纳入符合入组标准的24例心理亚健康及18例健康对照受试者,采集空腹晨尿经预处理后进行核磁共振氢谱法(1H-nuclear magnetic resonance spectroscopy,1H-NMR)测试。采用单变量、多元统计及随机森林算法对数据运算分析,以筛选造成组间差异的主要代谢物,发现脯氨酸、苯丙氨酸、葫芦巴碱、酮戊二酸、丙氨酸、柠檬酸、吲哚硫酸酚、乳酸、肌酸、牛磺酸及丙三醇11种代谢物的组间浓度表现出显著性差异(P0.05,0.01),提示可作为候选诊断生物标志物。进一步结合受试者工作曲线(receiver operating characteristic curve,ROC)及曲线下面积(area under the curve,AUC)对其作为诊断生物标志物组的能力进行评价。结果显示,单一生物标志物对心理亚健康的诊断能力较低,而将柠檬酸、葫芦巴碱及丙三醇3种物质联合应用后诊断能力明显提升(AUC=0.8)。上述结果提示,柠檬酸、葫芦巴碱及丙三醇3种候选生物标志物,可作为生物标志物组用于心理亚健康的诊断。生物标志物组的发现及其应用对于提高心理亚健康诊断与识别的准确度具有重要意义。     

阿尔茨海默病体外诊断纳米技术

阿尔茨海默病(Alzheimer's disease,AD)是一种最常见的神经退行性疾病.AD的精准诊断技术,特别是早期诊断技术是临床亟需的.近年来,以生物标志物为基础的非侵入性体外诊断技术发展迅速,特别是利用纳米材料和纳米技术的高表面活性、独特的光电特性、生物相容性好、易于表面修饰、小型化、集成化等特点,发展了新型的...     

结直肠癌无创筛查技术研究进展

结直肠癌和高危腺瘤的早期诊断及治疗,能够大大降低其死亡率。因而,在临床普及结直肠癌筛查有助于遏制该疾病的危害。目前,结肠镜检查是结直肠癌诊断的金标准,但该方法需要肠道准备,且具有侵入性,易造成肠道穿孔等缺点,导致患者依从性差,不利于其作为大规模筛查技术推广实施。近年来非侵入性的结直肠癌诊断方法发展迅速,这类方法主要通过检测粪便或血液样本中与结直肠癌发生相关的生物标志物,为结直肠癌的无创筛查提供了可能。但由于粪便、血液样本的成分复杂,对其中生物标志物的检测技术仍然在不断研究和完善中。本文分别从基于粪便样本和血液样本的检测技术两方面入手,探讨了近年来结直肠癌无创筛查技术的研究进展。     

昆虫多巴胺及其受体的研究进展及展望

多巴胺(dopamine,DA)是一种重要的神经递质,通过特异地结合其相关的多巴胺受体(dopamine receptors,DARs)发挥作用。昆虫DARs可分为D1-like DARs,D2-like DARs和多巴胺/蜕皮激素受体(dopamine/ecdysteroid receptor,DopEcR)。D1-like DARs包含两种亚型即DOP1和DOP2,都能偶联G s蛋白引起胞内cAMP上升,且DOP2还能偶联G q蛋白引起胞内Ca 2+浓度升高;D2-like DARs只有一种亚型DOP3,偶联G i蛋白导致胞内cAMP下降;DopEcR可以同时被DA和蜕皮激素激活。本文综述了近年来关于昆虫DA的调控、多巴胺神经元、DARs的药理学特性及生理功能等方面的研究进展。DA合成、转运和降解过程中的基因调控昆虫的多种表型,如表皮黑化、翅的颜色和图案等。DA在多巴胺神经元中合成和释放,不同类型的多巴胺神经元参与调控不同的功能。随着近年来单细胞测序和DA实时成像技术的兴起,这将有利于进一步探讨特异神经元的功能。不同昆虫DARs的激动剂和拮抗剂活性存在很大异同,这些药理学差异将为以昆虫DARs为作用靶标开发高效选择性杀虫剂提供重要依据。DARs参与调控昆虫的多种生理与行为过程,如取食、学习、记忆、遗忘、求偶、交配、睡眠及觉醒等。随着CRISPR/Cas9技术在不同昆虫中成功地应用,以及结合模式昆虫黑腹果蝇中丰富的遗传学操作手段,这些都将有利于精准解析DARs的功能。     

重组干扰素α-2b栓治疗宫颈人乳头瘤病毒亚临床感染的疗效及安全性观察

目的探讨重组干扰素α-2b栓治疗宫颈人乳头瘤病毒（HPV）亚临床感染的疗效及安全性观察。方法选择宫颈HPV亚临床感染患者80例,随机分为观察组与对照组。观察组患者予以重组人干扰素α-2b栓阴道内放置,每晚1枚,隔日1次,10d为1个疗程,连用3个疗程。对照组患者予以双唑泰栓阴道塞入,每晚1枚,隔日1次,10d为1个疗程,连用3个疗程。观察并比较两组患者治疗后的临床疗效及药物不良反应情况。结果治疗3个疗程后,观察组患者临床总有效率为95．0％,明显高于对照组的80．0％（X^2=4．11,P〈0．05）。对照组与观察组治疗期间分别出现不良反应4例和2例,均为阴道瘙痒,症状较轻微,未发生严重的药物不良反应。两组患者治疗期间不良反应发生率比较差异无统计学意义（X^2=0．18,P〉0．05）。结论重组干扰素α-2b栓治疗宫颈HPV亚临床感染的疗效较肯定,具有不良反应轻,安全性较好,使用方便等优点。     

Reciprocal modulation of function between the D1 and D2 dopamine receptors and the Na+,K+-ATPase

It is well documented that dopamine can increase or decrease the activity of the Na+,K+-ATPase (NKA, sodium pump) in an organ-specific fashion. This regulation can occur, at least partially, via receptor-mediated second messenger activation and can promote NKA insertion or removal from the plasma membrane. Using co-immunoprecipitation and mass spectrometry, we now show that, in both brain and HEK293T cells, D1 and D2 dopamine receptors (DARs) can exist in a complex with the sodium pump. To determine the impact of NKA on DAR function, biological assays were conducted with NKA and DARs co-expressed in HEK293T cells. In this system, expression of NKA dramatically decreased D1 and D2 DAR densities with a concomitant functional decrease in DAR-mediated regulation of cAMP levels. Interestingly, pharmacological inhibition of endogenous or overexpressed NKA enhanced DAR function without altering receptor number or localization. Similarly, DAR function was also augmented by small interfering RNA reduction of the endogenous NKA. These data suggest that, under basal conditions, NKA negatively regulates DAR function via protein-protein interactions. In reciprocal fashion, expression of DARs decreases endogenous NKA function in the absence of dopamine, implicating DAR proteins as regulators of NKA activity. Notably, dopamine stimulation or pertussis toxin inhibition of D2 receptor signaling did not alter NKA activity, indicating that the D2-mediated decrease in NKA function is dependent upon protein-protein interactions rather than signaling molecules. This evidence for reciprocal regulation between DARs and NKA provides a novel control mechanism for both DAR signaling and cellular ion balance.     

双歧杆菌三联活菌散剂联合四磨汤治疗儿童厌食症的疗效

目的探讨双歧杆菌三联活菌散剂联合四磨汤治疗儿童厌食症的疗效。方法选取儿童厌食症患者80例,随机分为观察组和对照组。两组患儿均予以补锌、健脾和纠正不良饮食习惯等常规治疗。观察组患儿予以双歧杆菌三联活菌散剂联合四磨汤治疗,对照组患儿予以单纯的四磨汤治疗,两组疗程8周。观察两组患儿治疗前后腹部皮下脂肪厚度的变化,并比较其临床疗效及药物不良反应。结果治疗8周后,两组患者腹部皮下脂肪厚度均较前明显增加（P〈0.05或P〈0.01）,且观察组增加的幅度明显高于对照组（P〈0.05）;同时观察组患儿的总有效率明显高于对照组（95.0%vs 80.0%）（χ2=4.11,P〈0.05）,两组患儿治疗期间未发生明显的药物不良反应。结论双歧杆菌三联活菌散剂联合四磨汤治疗儿童厌食症的疗效确切,能明显增强患儿的食欲,增加皮下脂肪厚度,安全性好。     

Biomarkers for cardiovascular disease: challenges and future directions

The accurate diagnosis and prevention of cardiovascular disease (CVD) is an important public health goal. Although clinical characteristics such as age and gender are well-established risk factors for CVD, such features are not sufficient to identify all patients at risk. Cardiovascular biomarkers have the potential to augment clinical risk stratification by aiding in screening, diagnosis and assessment of prognosis. However, most current biomarkers have only modest predictive value, and there is a need to identify additional biomarkers from new biological pathways. The availability of platforms for profiling DNA, RNA, proteins and metabolites in clinical specimens has facilitated the 'unbiased' search for new biomarkers, which can now be tested in a clinical setting. This review highlights recent developments in the field of cardiovascular biomarkers and describes the use of new technologies for the identification of biomarkers.     

Sphingolipids as New Biomarkers for Assessment of Delayed-Type Hypersensitivity and Response to Triptolide

Background

Hypersensitivity diseases are associated with many severe human illnesses, including leprosy and tuberculosis. Emerging evidence suggests that the pathogenesis and pathological mechanisms of treating these diseases may be attributable to sphingolipid metabolism.

Methods

High performance liquid chromatography-tandem mass spectrometry was employed to target and measure 43 core sphingolipids in the plasma, kidneys, livers and spleens of BALB/c mice from four experimental groups: control, delayed-type hypersensitivity (DTH) model, DTH+triptolide, and control+triptolide. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify potential biomarkers associated with variance between groups. Relationships between the identified biomarkers and disease markers were evaluated by Spearman correlation.

Results

As a treatment to hypersensitivity disease, triptolide significantly inhibit the ear swelling and recover the reduction of splenic index caused by DTH. The sphingolipidomic result revealed marked alterations in sphingolipid levels between groups that were associated with the effects of the disease and triptolide treatment. Based on this data, 23 potential biomarkers were identified by OPLS-DA, and seven of these biomarkers correlated markedly with the disease markers (p<0.05) by Spearman correlation.

Conclusions

These data indicate that differences in sphingolipid levels in plasma and tissues are related to DTH and treatment with triptolide. Restoration of proper sphingolipid levels may attribute to the therapeutic effect of triptolide treatment. Furthermore, these findings demonstrate that targeted sphingolipidomic analysis followed by multivariate analysis presents a novel strategy for the identification of biomarkers in biological samples.     

Examination of protein adsorption in fluidized bed and packed bed columns at different temperatures using frontal chromatographic methods

The influences of various experimental parameters on the dynamic adsorption capacity (DAC) and the dynamic adsorption rate (DAR) of a biomimetic affinity silica-based adsorbent in fluidized and packed bed columns operated under plug flow conditions and at different temperatures have been investigated with different inlet concentrations of hen egg white lysozyme (HEWL) and human serum albumin (HSA). The DACs as well as the DARs of both the fluidized and packed beds were examined at 10% saturation (i.e., at the QB value) and the experimental data compared with the corresponding data obtained from batch equilibrium adsorption procedures. Parameters examined included the fluid superficial velocity and protein concentration and their effect on the binding capacity and column efficiency. Consistent with various results reported from this and other laboratories on the behavior of biospecific affinity adsorbents derived from porous silica and zirconia particles, adsorbents prepared from Fractosil 1000 were found to exhibit appropriate rheological characteristics in fluidized bed systems under the experimental conditions. Moreover, changes in temperature resulted in a more significant effect on the breakthrough profiles of HSA compared to HEWL with the immobilized Cibacron Blue F3G-A with Fractosil 1000 adsorbent. This result suggests that temperature effects can possibly be employed profitably in some processes as part of a strategy to enhance column performance with fluidized bed systems for selective recovery of target proteins. At relatively low superficial velocities of the feed, the DARs with HEWL and HSA were similar for both the fluidized and packed bed column systems, whereas, at high superficial velocities, the DARs for these proteins were larger with the packed bed columns.