水热法制备质谱专用提取多肽的金属螯合纳米磁珠

应用水热法合成质谱专用提取血清多肽的金属螯合纳米磁珠, 可用于质谱对血清中多肽分布情况的研究, 获得血清多肽谱. 对磁珠进行透射电镜(TEM), 原子力显微镜(AFM)和傅立叶红外光谱FT-IR的表征, 显示该粒子粒径在70～90 nm. 并通过质谱验证该金属螯合磁珠能有效提取血清中的多肽, 该磁珠为质谱进行疾病诊断解决样品制备的技术难题, 具有广阔应用前景.     

人肝癌细胞系HLE细胞表面抗原多肽的纳升电喷雾串联质谱从头测序分析

肿瘤细胞表面的抗原多肽能够被细胞毒T淋巴细胞特异性识别而引起免疫应答,因此有可能用于研制基于多肽的抗肿瘤疫苗。用弱酸将人肝癌细胞系HLE细胞表面抗原多肽和人正常肝细胞表面多肽洗脱后,经RP-HPLC分离,选择HLE细胞表面特异性多肽进行纳升电喷雾串联质谱(nanoESI-MS/MS)测序,共测定5个色谱峰中的20个多肽序列,分子量分布范围为1000~2000 Da。借助M asSeq软件分析出其中12个多肽的序列。经数据库查寻,其中的3个肽段分别来自钙调节蛋白、核蛋白S19和伴侣蛋白10。这些多肽的生物学功能及与肿瘤的关系值得深入研究。该研究表明nanoESI-MS/MS是测定微量混合多肽序列的最有效方法。     

毛细管电泳-发光二极管诱导荧光检测法分析肝硬化及正常人血清蛋白质的差异

应用毛细管电泳技术建立了人血清蛋白质毛细管电泳分析方法,对比了正常人与肝硬化患者血清蛋白质含量的差异。使用异硫氰酸荧光素(FITC)进行荧光标记,并考察光源与滤光片的波长、缓冲液的浓度、pH值、分离电压对分离效果的影响。建立了毛细管电泳-发光二极管诱导荧光法检测肝硬化患者血清及正常人血清蛋白质的方法;通过分离检测,获得了两者的蛋白质电泳谱图。经分析,在肝硬化患者的血清蛋白质中比在正常人的血清蛋白质中多检测到2个蛋白质峰,可能是和肝硬化发生相关的特异蛋白质;肝硬化患者与正常人的血清蛋白质电泳谱图的差异有统计学意义。该方法能实现人血清蛋白质的分离,可为临床诊断肝硬化做参考。     

核酸适配体功能化近红外量子点结合流式细胞术快速检测白血病细胞

利用作为肿瘤细胞识别分子的核酸适配体(Aptamer)的高特异性和高亲和力以及作为信号报告单元的近红外量子点(QDs)的高荧光发射强度和低生物背景干扰特性,构建了一种基于Aptamer功能化近红外QDs的新型纳米荧光探针,并进一步结合流式细胞术在单细胞荧光分析方面的高通量、简便和快速等优势,建立了一种检测白血病细胞的新方法.以基于Cell-SELEX(Cell-based systematic evolution of ligands by exponential enrichment)技术针对CCRF-CEM人急性白血病细胞筛选的特异性Aptamer Sgc8c为模型,构建了Sgc8c-QDs探针,其仅需与细胞样品培育30 min即可实现对缓冲液和血清中靶细胞的简单、快速和高特异性检测.与传统荧光染料标记技术相比,该方法不仅大大提高了分析灵敏度,还显示出对血清等复杂生物样品的高适用性和优良检测性能.鉴于Cell-SELEX技术在其它白血病细胞Aptamer筛选领域的应用潜力,该方法有望作为一种通用技术在白血病诊断及预后监测等方面发挥重要作用.     

基于质谱技术研究黑色素瘤血清特异性O-糖链

对接种和未接种B16黑色素瘤细胞的C57小鼠进行血清O-糖链比较糖组学研究,寻找黑色素瘤血清特异性O-糖链.小鼠血清10μL,β-消除反应释放O-糖链.反应混合物经石墨化炭黑固相萃取小柱(GCC SPE)分离纯化后,用于MALDI-Qit-TOF-MS分析.通过Launchpad软件采集并输出质谱数据,MATLAB进行数据解析,找到了10个稳定出现的差异糖链质谱峰.利用串联质谱分析了其中5个主要差异糖链的结构.     

用反相液相色谱分离多肽时峰容量与色谱柱长度关系的研究

在蛋白质组学研究中,多肽混合物的有效分离对蛋白质鉴定和蛋白质之间相互作用的研究起着决定性的影响。基于此,用反相液相色谱研究了在两个不同长度的色谱柱上分离多肽混合物时色谱柱长度与峰容量的关系,同时考察了梯度洗脱时间对峰容量和峰宽的影响。实验结果表明,色谱柱长度对峰容量有显著的影响,而延长梯度洗脱时间不仅可以增加峰容量,而且可以增加峰宽。这说明用毛细管液相色谱 串联质谱联用方法对多肽混合物进行分离鉴定时,采用较长的色谱柱和较长的梯度洗脱时间有利于对更多的多肽进行分析鉴定。     

儿童急性白血病骨髓浸润的MRI与MRS分析

目的 研究儿童急性白血病在发病初期、化疗缓解期及复发期骨髓MRI及MRS成像。方法 研究大连市妇女儿童医疗中心2015年2月至2016年8月间30例儿童(2~16岁)急性白血病患者的MRI平扫及MRS成像特征。急性淋巴细胞白血病25例,急性非淋巴细胞白血病5例;早期组24例,化疗缓解组21例,复发组2例,17例分别在发病初期、化疗缓解期或复发期行MRI检查。对照组为30例同年龄段正常儿童志愿者。结果 急淋初始发病时骨髓在T1信号减低,STIR示骨髓信号较均匀增高;MRS显示发病初期组脂峰消失或地平,水峰高耸。化疗缓解组骨髓腔信号在T1较初发组信号增高;STIR示骨髓信号减低;MRS脂峰复现。早期组与化疗缓解组比较、早期组与对照组之间骨髓FF%有显著性差异(P0.05),而化疗缓解组与对照组比较,无显著性差异(P0.05)。结论 骨髓MRI及MRS作为一种无创性的检查工具,对儿童急性白血病骨髓状态及评价预后具有价值。     

IEX/RP二维液相色谱中弱保留组分收集模式的构建及系统评价

以十通阀和捕集柱接口形式,构建了弱阴离子交换/反相(WAX/RP)二维液相色谱分离系统.通过将第一维离子交换色谱分析中的前部集中洗脱出的弱保留组分收集后单独分析,剩余样品进一步采用二维液相色谱分析,可以有效避免第二维色谱柱对特殊样品局部集中流出导致的第二维分离超柱容量问题,提高了系统的整体分离能力.使用4种蛋白胰蛋白酶酶解后的多肽样品评价该系统,在不分流的系统中共检测到115个峰.对第一维分离的前15 min分流后得到的组分单独分析,共识别出58个峰,后续的二维分离中共得到78个峰.2种方法相比,第二种方法检测峰数增加了21个,一些低丰度的组分在弱保留组分收集后被识别.     

微流控芯片中磁免疫荧光法检测新城疫病毒

建立了一种基于微流控芯片的磁免疫荧光方法,并用于新城疫病毒(NDV)的高灵敏检测。该方法基于抗原抗体免疫识别作用,采用免疫磁珠实现了对NDV的高效捕获和分离,并利用量子点的荧光作为检出信号,提高方法的灵敏度。在优化条件下,NDV的检测限为1ng/mL,线性范围为1～10ng/mL。利用免疫磁珠捕获分离病毒,可用于家禽肝脏、肺、粪便等实际样品中病毒的检测。该方法可以在1h内完成对NDV的检测,操作简单、特异性强,灵敏度高,重现性好。本文为NDV的检测提供了一种快速、准确的方法。     

基于超高效液相色谱-高分辨质谱的多肽组学技术用于人参不同部位多肽的差异分析

采用基于超高效液相色谱-高分辨质谱的多肽组学技术对人参主根、支根、须根和芦头的多肽谱进行全面分析，旨在评价人参不同形态区域多肽表达的异同。本研究共表征62个数据库中已收录的人参多肽。结果表明，人参不同部位均富含多肽类成分。多肽组学研究发现，人参主根和支根、芦头与须根之间多肽含量具有显著差异，从鉴定到的多肽中共发现25个稳定表达的已知潜在多肽标志物。其中多肽种类及含量在主根与其他部位间差异最显著，为主根与非主根药效差异研究提供了新思路。本研究揭示了人参多肽结构多样性及人参不同部位人参多肽表达的异同，对人参化学特征评价具有重要意义，为人参质量控制和合理应用提供了化学依据。     

Decoration of silver nanoparticles on multi-walled carbon nanotubes: Investigation of its anti-acute leukemia property against acute myeloid leukemia and acute T cell leukemia

Recently, the development of carbon nanocomposites composed of carbon nanotubes and metal nanoparticles has attracted many interests because of their large potential for technological applications such as catalysts, sensors, biomedicine, and disinfection. In the present study, we described a simple chemistry method to synthesize multi-walled carbon nanotubes (MWCNTs) decorated with silver nanoparticles (Ag-NPs). Also, we investigated the antioxidant and anti-acute leukemia activities against acute myeloid leukemia and acute T cell leukemia cell lines. Ag NPs-MWCNTs were characterized and analyzed using common nanotechnology techniques including transmission electron microscopy (TEM), X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDS), field emission-scanning electron microscopy (FE-SEM) and elemental mapping analysis. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) test was performed to assess the antioxidant capacities of AgNO₃, MWCNTs, and Ag NPs-MWCNTs. It revealed similar antioxidant potentials for Ag NPs-MWCNTs and butylated hydroxytoluene. In MTT assay, Ag NPs-MWCNTs had very low cell viability (very high anti-acute leukemia properties) dose-dependently against 32D-FLT3-ITD (Acute myeloid leukemia cell line), Human HL-60/vcr (Acute myeloid leukemia cell line), Jurkat, Clone E6–1 (Acute T cell leukemia cell line), and J.RT3-T3.5 (Acute T cell leukemia cell line) without any cytotoxicity on human umbilical vein endothelial cell line (HUVEC; Normal cell line). In conclusion, the synthesized Ag NPs-MWCNTs revealed excellent antioxidant and cytotoxicity activities against acute myeloid leukemia and acute T cell leukemia cell lines in a dose depended manner. After confirming in the in vivo and clinical trials, these nanoparticles can be administrated in humans for the treatment of acute leukemia especially acute myeloid leukemia and acute T cell leukemia.     

Gas Chromatographic Determination of Ethanol in Rabbit Arterial Serum

Abstract

A glc method utilizing a flame ionization detector was developed for determining ethanol (I) in rabbit arterial serum. Addition of triton X-100 solution containing the internal standard, acetonitrile (II) to rabbit serum was followed by direct injection glc analysis using Porapak Q as the stationary phase. Ethanol was clearly separated from its major metabolite acetaldehyde (III) although the assay was not suitable for determination of III. Accuracy and precision of the method was demonstrated over the concentration range anticipated in acute intravenous dosing of I to rabbits. The method was evaluated in a rabbit which received three different intravenous infusion doses of I at two week intervals and found to be applicable to arterial serum analysis.     

54例白血病患者血清硒含量分析

以催化极谱法测定了５４例白血病患者血清硒含量。结果表明，急淋、急粒患者血清硒含量均低于正常对照组，慢粒及急、慢粒经治疗缓解者血清硒含量与正常对照组间均无显著性差异，提示低硒状态只存在于急性患者，且随病情缓解后血硒水平趋于正常，慢粒与血硒含量没有明显的相关关系。     

Formulation of a novel anti-leukemia drug and evaluation of its therapeutic effects in comparison with Cytarabine

In the recent research, we investigated the application of gold nanoparticles green-synthesized by Alhagi maurorum aqueous extract in the treatment of several types of leukemia, i.e. acute T cell leukemia, acute lymphoblastic leukemia, and acute myeloid leukemia. Different techniques such as transmission electron microscopy (TEM), fourier-transform infrared spectroscopy (FTIR), and ultraviolet–visible spectroscopy analysis were used to characterize AuNPs. The TEM images show a spherical morphology for AuNPs with the range size of 21 to 59 for the synthetic nanoparticles. In the antioxidant test, the IC50 of AuNPs and butylated hydroxytoluene (BHT) against DPPH free radicals were 117 and 87 µg/mL, respectively. In the oncological part of the recent study, the treated cells with AuNPs and Cytarabine were assessed by MTT assay for 48 h about the cytotoxicity and anti-leukemia properties on normal (HUVEC) and leukemia cell lines i.e., acute myeloid leukemia (Human HL-60/vcr and 32D-FLT3-ITD), acute lymphoblastic leukemia (MOLT-3 and TALL-104), and acute T cell leukemia (J.RT3-T3.5 and Jurkat, Clone E6-1). The IC50 of AuNPs were 242, 297, 383, 207, 234, and 218 µg/mL against acute myeloid leukemia (Human HL-60/vcr and 32D-FLT3-ITD), acute lymphoblastic leukemia (MOLT-3 and TALL-104), and acute T cell leukemia (J.RT3-T3.5 and Jurkat, Clone E6-1) cell lines, respectively. In addition, the IC50 of Cytarabine were 117, 113, 145, 119, 131, and 135 µg/mL against acute myeloid leukemia (Human HL-60/vcr and 32D-FLT3-ITD), acute lymphoblastic leukemia (MOLT-3 and TALL-104), and acute T cell leukemia (J.RT3-T3.5 and Jurkat, Clone E6-1) cell lines, respectively. The viability of malignant leukemia cell line reduced dose-dependently in the presence of AuNPs and Cytarabine.     

白血病患儿体内铜锌代谢的动态观察

对白血病患儿体内血清锌、血清铜浓度以及血清铜/锌比值作了初步的观测.在41例白血病患儿和33例正常对照儿童中,观察到在明确诊断化疗前,41例白血病患儿中有20例患儿的血清锌浓度低于正常低限,他们的平均血清锌浓度为12.3±3.4 μmol/L,明显低于对照组18.3±4.0 μmol/L.白血病患儿的血清铜/锌比值为1.40±0.38,明显高于正常儿童1.19±0.25.经治疗后,有12例患儿获缓解并继续随访至发病后6～8周,平均血清锌浓度由11.4±2.2μmol/L,提高到13.7±2.1μmol/L,血清铜/锌比值由1.51±0.36,下降到1.29±0.23.说明白血病患儿体内存在着锌代谢的异常.     

DNA-protein complexes

Sera of patients with different types of leukemia and acquired immune deficiency syndrome (AIDS) have been examined for the presence of the anti-DNA antibodies. DNA-hydrolyzing activity of antibodies was detected in the sera of patients with chronic lymphoid leukemia (CLL), pre-B-cell acute lymphoid leukemia (pre-B-All), acute myeloleukosis (AML), and AIDS in stages III and IV of the disease. In immunofluorescence tests, the DNA-hydrolyzing antibodies reacted preferentially with proliferating cell nuclei compared with resting cells. A 35-kDa factor was identified as the target for the DNA antibodies in the cell nuclei. The DNA-hydrolyzing antibody fraction from the serum of an AIDS patient crossreacted with HIV I virus proteins gp160, gp120, and p65.     

Serum thymidine kinase activity of various cancer and HBV positive liver diseases

Clinical utility of determination of serum deoxythymidine kinase (TK) activity is described. It is well known that elevated TK level is observed in leukemia and other malignant diseases, or some viral infectious diseases. The TK activity was assayed on normal subjects, hepatitis B virus (HBV) positive liver diseases and various cancer by a newly developed high sensitive method, radio enzyme assay (REA) utilizing 125I-iododeoxyuridine as the substrate. Measurement of TK activity by the REA is revealed to be useful for "the marker of DNA metabolism anomaly" in leukemia, etc.     

Green formulation,chemical characterization and anti-acute leukemia effects of vanadium nanoparticles containing Foeniculum vulgare extract

In this study, vanadium nanoparticles (VNPs) were green synthesized using Foeniculum vulgare extract. VNPs were characterized using chemical analysis techniques including FT-IR, XRD, FE-SEM, TEM and EDS. The microscopy techniques revealed a spherical morphology for the particles with size less than 50 nm. According to XRD data V₂O₅ was confirmed for VNPs. Maybe significant anti-human acute leukemia potentials of the synthesized nanoparticles against common human acute leukemia cell lines are linked to their antioxidant activities. MTT assay was used on common acute leukemia cell lines i.e., 32D-FLT3-ITD, MOLT-3 and Jurkat, Clone E6-1 to survey the cytotoxicity and anti-acute leukemia effects of the synthesized nanoparticles. The synthesized nanoparticles had very low cell viability and high anti-acute leukemia activities dose-dependently against 32D-FLT3-ITD, MOLT-3 and Jurkat, Clone E6-1 cell lines without cytotoxicity on the normal cell line (HUVEC). To determine the antioxidant properties of the synthesized nanoparticles, the DPPH test was used in the presence of butylated hydroxytoluene as the positive control. The IC₅₀ of VNPs were 25, 33 and 26 µg/mL against 32D-FLT3-ITD, MOLT-3 and Jurkat, Clone E6-1 cell lines, respectively. The synthesized nanoparticles inhibited half of the DPPH molecules in the concentration of 28 µg/mL.     

Monolith and coating enzymatic microreactors of L-asparaginase: kinetics study by MCE-LIF for potential application in acute lymphoblastic leukemia (ALL) treatment

The study of enzyme immobilization using an extracorporeal shunt system is essential to eliminate the side effects of L-asparaginase (L-Asnase; including hepatic toxicity, allergic reaction, pancreatitis, central nervous system toxicity and decreased synthesis of blood clotting factors) when it was applied as an anticancer drug given directly to patients by injection. Thus, the novel monolith and coating enzymatic reactors of L-asparaginase were provided in this assay and a microchip electrophoresis-laser induced fluorescence (MCE-LIF) method was set up for the enzyme kinetics study. The enzymatic reactors would be a promising in vitro therapeutic method in an extracorporeal shunt system for acute lymphoblastic leukemia (ALL) treatment. For the first time, L-asparaginase was covalently bound to the polymer monolith and coating in the capillary and the activity characteristics of these enzymatic microreactors have been probed by Michaelis-Menten kinetic constants. Meanwhile, the D,L-amino acids were chirally separated using microchip electrophoresis with a laser induced detector and D,L-aspartic acid (D,L-Asp) were tested for the L-asparaginase enzymatic reactor kinetics study. Furthermore, human serum adding with L-asparagine (L-Asn) as the sample was hydrolyzed by the enzymatic microreactors. The results demonstrated that the developed enzymatic microreactor of L-asparaginase would be a potential therapeutic protocol for ALL treatment.     

Nylon tube-immobilized creatinine iminohydrolase and glutamate dehydrogenase in serum and urine creatinine analysis

Immobilized enzyme nylon-tube reactors incorporating creatinine iminohyrolase (CI) and glutamate dehydrogenase (GDH) were used to assay creatinine in serum and urine. Optimum substrate concentrations for the assay were determined. The reactors were incorporated into a continuous flow system for creatinine analysis. The method was evaluated with respect to linearity, sample interaction, precision, accuracy, and analytical recovery. Comparison studies were carried out with a standard Jaffé method and the effect of interfering substances was investigated. From the results obtained, it was concluded that the assay was suitable as a simple, reliable, and specific method for serum and urine creatinine determinations.