硒化壳聚糖抑制人早幼粒白血病细胞增殖的研究

为探讨硒化壳聚糖对体外培养人早幼粒白血病细胞增殖的抑制作用,用SRB法和集落形成法检测了药物对细胞增殖的抑制作用,流式法检测了细胞周期阻断作用。结果表明,25、50、100mg／L硒化壳聚糖作用HL60细胞48h对细胞有增殖抑制作用（P〈0．01）;50、100mg／L硒化壳聚糖作用细胞48h后,G0／G1期细胞数较对照组增加了14．9％-22．0％（P〈0．05）,S期细胞减少了14．3％～20．1％（P〈0．05）。可见硒化壳聚糖对人早幼粒白血病细胞增殖具有抑制作用。     

Apoptotic and Cell Cycle Effects of Triterpenes Isolated from Phoradendron wattii on Leukemia Cell Lines

Current antineoplastic agents present multiple disadvantages, driving an ongoing search for new and better compounds. Four lupane-type triterpenes, 3α,24-dihydroxylup-20(29)-en-28-oic acid (1), 3α,23-dihydroxy-30-oxo-lup-20(29)-en-28-oic acid (2), 3α,23-O-isopropylidenyl-3α,23-dihydroxylup-20(29)-en-28-oic acid (3), and 3α,23-dihydroxylup-20(29)-en-28-oic acid (4), previously isolated from Phoradendron wattii, were evaluated on two cell lines of chronic (K562) and acute (HL60) myeloid leukemia. Compounds 1, 2, and 4 decreased cell viability and inhibit proliferation, mainly in K562, and exhibited an apoptotic effect from 24 h of treatment. Of particular interest is compound 2, which caused arrest in active phases (G2/M) of the cell cycle, as shown by in silico study of the CDK1/Cyclin B/Csk2 complex by molecular docking. This compound [3α,23-dihydroxy-30-oxo-lup-20(29)-en-28-oic acid] s a promising candidate for incorporation into cancer treatments and deserves further study.     

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 Aptamers in the Detection of Leukemia Cells by the Thickness Shear Mode Acoustics Method

By using the thickness shear mode acoustics method (TSM) and single-molecule force spectroscopy (SMFS) we studied the interactions between DNA aptamers (sgc8c) specific to the protein tyrosine kinase 7 (PTK7), which is localized in the membranes of leukemia lymphoblastics (MOLT-4), and lymphocyte (Jurkat) cell lines, as well with PTK7-negative U266 myeloid leukemia cells. The TSM method allowed the development of a highly sensitive, label-free biosensor for the detection leukemia cells with a limit of detection of (195±20) cells/mL. SMFS approved the high selectivity of the sgc8c aptamers to the PTK7 receptors at the cell surface and allowed determining the binding probability of the aptamers to the PTK7 receptors at different cell lines.     

Cephalotaxine Inhibits the Survival of Leukemia Cells by Activating Mitochondrial Apoptosis Pathway and Inhibiting Autophagy Flow

Cephalotaxine (CET) is a natural alkaloid with potent antileukemia effects. However, its underlying molecular mechanism has not been well understood. In this study, we verified that CET significantly inhibited the viability of various leukemia cells, including HL-60, NB4, Jurkat, K562, Raji and MOLT-4. RNA-sequencing and bioinformatics analysis revealed that CET causes mitochondrial function change. Mechanism research indicated that CET activated the mitochondrial apoptosis pathway by reducing the mitochondrial membrane potential, downregulating anti-apoptotic Bcl-2 protein and upregulating pro-apoptotic Bak protein. In addition, the autophagy signaling pathway was highly enriched by RNA-seq analysis. Then, we found that CET blocked the fluorescence colocation of MitoTracker Green and LysoTracker Red and upregulated the level of LC3-II and p62, which indicated that autophagy flow was impaired. Further results demonstrated that CET could impair lysosomal acidification and block autophagy flow. Finally, inhibiting autophagy flow could aggravate apoptosis of HL-60 cells induced by CET. In summary, this study demonstrated that CET exerted antileukemia effects through activation of the mitochondria-dependent pathway and by impairing autophagy flow. Our research provides new insights into the molecular mechanisms of CET in the treatment of leukemia.     

Expanding the Study of the Cytotoxicity of Incomptines A and B against Leukemia Cells

Heliangolide-type sesquiterpene lactones (HTSLs) are phytocompounds with several pharmacological activities including cytotoxic and antitumor activity. Both bioactivities are related to an α-methylene-γ-lactone moiety and an ester group on carbon C-8 in the sesquiterpene lactone (SL) structure. Two HTSLs, incomptines A (AI) and B (IB) isolated from Decachaeta incompta, were evaluated for their cytotoxic activity on three leukemia cell lines: HL-60, K-562, and REH cells. Both compounds were subjected to a molecular docking study using target proteins associated with cancer such as topoisomerase IIα, topoisomerase IIβ, dihydrofolate reductase, methylenetetrahydrofolate dehydrogenase, and Bcl-2-related protein A1. Results show that IA and IB exhibit cytotoxic activity against all cell lines used. The CC₅₀ value of IA was 2–4-fold less than etoposide and methotrexate, two anticancer drugs used as positive controls. The cytotoxic activity of IB was close to that of etoposide and methotrexate. The molecular docking analysis showed that IA and IB have important interaction on all targets used. These findings suggest that IA and IB may serve as scaffolds for the development of new treatments for different types of leukemia.     

基于超高效液相色谱-离子阱-飞行时间质谱联用的白血病细胞及耐药细胞非靶标代谢组学研究

建立了一种基于超高效液相色谱-离子阱-飞行时间质谱(UPLC-IT-TOF MS)联用的非靶标代谢组学方法,研究了人慢性髓系白血病细胞及其阿霉素耐药细胞(K562和K562-ADM)之间的代谢物差异。考察了细胞破碎方法、提取溶剂体系和溶剂体积对细胞代谢物提取效果的影响,实验发现采用超声破碎法,以800μL的80%甲醇提取时可得到较多的代谢特征峰,方法重复性和稳定性较高。将建立的方法应用于K562及K562-ADM细胞的代谢组差异研究,对所得数据进行主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA)。结果显示2种细胞的代谢特征存在显著差异,并发现40个对分类有显著贡献的代谢物,差异代谢物主要参与氨基酸代谢、鞘脂代谢、嘌呤代谢和嘧啶代谢等通路。该方法可用于K562和K562-ADM细胞的代谢组差异研究,可望为白血病细胞耐药性的代谢组学研究提供帮助。     

Determination of Imatinib in the Blood Cells of Chronic Myelogenous Leukemia Patients by Ion-Trap Mass Spectrometry

Imatinib mesylate is a standard first-line therapy for patients with chronic myelogenous leukemia. However, there is still a significant proportion of these patients who reflect sub-optimal responses or fail imatinib therapy. Knowledge of the distribution within the studied system (e.g., peripheral blood) may be of high importance for understanding the principles of drug action and possible patient resistance to treatment. Intracellular or more precisely cell-associated, imatinib concentrations in patients, were shown to be higher compared to those in plasma, but still only limited data related to the methodology aspects of cell-associated concentrations are available. Herein is presented an assessment of the cell-associated imatinib determination assay by mass spectrometry. Three approaches were evaluated to isolate cells from the peripheral blood of chronic myelogenous leukemia patients. Erythrocyte lysis was found to cause substantial leakage of cell-associated imatinib in the first step. Selected alternative procedures utilizing density gradients did not affect the cell-associated imatinib concentration significantly. Cell isolates were subjected to flow cytometry which revealed differences in the population composition of peripheral blood cell isolates among individual patients indicating that the cell isolate composition should be addressed with the cell-associated imatinib concentration. The proposed approach may be utilized for the determination of intracellular concentration of imatinib and for other drugs in which the intracellular concentration plays a key role in the therapy.     

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。     

Dual Targeting of MDM4 and FTH1 by MMRi71 for Induced Protein Degradation and p53-Independent Apoptosis in Leukemia Cells

MDM2 and MDM4 are cancer drug targets validated in multiple models for p53-based cancer therapies. The RING domains of MDM2 and non-p53-binder MDM2 splice isoforms form RING domain heterodimer polyubiquitin E3 ligases with MDM4, which regulate p53 stability in vivo and promote tumorigenesis independent of p53. Despite the importance of the MDM2 RING domain in p53 regulation and cancer development, small molecule inhibitors targeting the E3 ligase activity of MDM2-MDM4 are poorly explored. Here, we describe the synthesis and characterization of quinolinol derivatives for the identification of analogs that are capable of targeting the MDM2-MDM4 heterodimer E3 ligase and inducing apoptosis in cells. The structure-activity-relationship (SAR) study identified structural moieties critical for the inhibitory effects toward MDM2-MDM4 E3 ligase, the targeted degradation of MDM4 and FTH1 in cells, and anti-proliferation activity. Lead optimization led to the development of compound MMRi71 with improved activity. In addition to accumulating p53 proteins in wt-p53 bearing cancer cells as expected of any MDM2 inhibitors, MMRi71 effectively kills p53-null leukemia cells, an activity that conventional MDM2-p53 disrupting inhibitors lack. This study provides a prototype structure for developing MDM4/FTH1 dual-targeting inhibitors as potential cancer therapeutics.     

Polyphenolic Flavonoid Compound Quercetin Effects in the Treatment of Acute Myeloid Leukemia and Myelodysplastic Syndromes

Flavonoids are ubiquitous groups of polyphenolic compounds present in most natural products and plants. These substances have been shown to have promising chemopreventive and chemotherapeutic properties with multiple target interactions and multiple pathway regulations against various human cancers. Polyphenolic flavonoid compounds can block the initiation or reverse the promotion stage of multistep carcinogenesis. Quercetin is one of the most abundant flavonoids found in fruits and vegetables and has been shown to have multiple properties capable of reducing cell growth in cancer cells. Acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) therapy remains a challenge for hematologists worldwide, and the outcomes for patients with both disorders continue to be poor. This scenario indicates the increasing demand for innovative drugs and rational combinative therapies. Herein, we discuss the multitarget effects of the flavonoid quercetin, a naturally occurring flavonol, on AML and MDS.     

白血病骨髓移植患者口腔黏膜病与微量元素锌的关系

为研究白血病骨髓移植患者全血微量元素锌与口腔黏膜病的关系，用原子吸收光谱法检测了正常对照组与白血病骨髓移患者预处理前及移植后骨髓空虚期全血微量元素锌的含量，并观察了白血病骨髓移植患者空黏膜病变情况。结果表明，正常对照组全血锌浓度与白血病骨髓移植患者预处理前全血锌浓度比较，差异无显著性（P＞0．05），而预处理前全血锌浓度与骨髓空虚期全血锌浓度有显著性差异（P＜0．01），白血病缓解后（预处理前）血锌接受正常水平，无口腔溃疡发生，而白血病骨髓移植患者处理后骨髓空虚期全血锌含量显著降低，并出现不同程度的口腔黏膜病变，说明预处理影响微量元素锌的代谢，微量元素锌的减少与白血病骨髓移植患者移植患者口腔黏膜病的发生有关。     

白血病骨髓移植病人全血微量元素铜的变化及其意义

为了研究白血病骨髓移植病人全血微量元素铜的变化及其意义 ,用原子吸收光谱法检测了正常对照组与白血病骨髓移植患者预处理前及移植后骨髓空虚期全血微量元素铜的含量。结果表明 ,正常对照组全血铜浓度与白血病骨髓移植患者预处理前全血铜浓度差异无显著性 (P >0 0 5 ) ,而与骨髓空虚期全血铜浓度有显著性差异 (P <0 0 5 ) ,说明白血病缓解后血铜接近正常对照组水平 ,而白血病骨髓移植患者预处理后骨髓空虚期全血铜含量高于正常对照组 ,说明预处理影响微量元素铜的代谢 ,本文就其影响机理进行了研究     

Co-Treatments of Edible Curcumin from Turmeric Rhizomes and Chemotherapeutic Drugs on Cytotoxicity and FLT3 Protein Expression in Leukemic Stem Cells

This study aims to enhance efficacy and reduce toxicity of the combination treatment of a drug and curcumin (Cur) on leukemic stem cell and leukemic cell lines, including KG-1a and KG-1 (FLT3⁺ LSCs), EoL-1 (FLT3⁺ LCs), and U937 (FLT3⁻ LCs). The cytotoxicity of co-treatments of doxorubicin (Dox) or idarubicin (Ida) at concentrations of the IC₁₀–IC₈₀ values and each concentration of Cur at the IC₂₀, IC₃₀, IC₄₀, and IC₅₀ values (conditions 1, 2, 3, and 4) was determined by MTT assays. Dox–Cur increased cytotoxicity in leukemic cells. Dox–Cur co-treatment showed additive and synergistic effects in several conditions. The effect of this co-treatment on FLT3 expression in KG-1a, KG-1, and EoL-1 cells was examined by Western blotting. Dox–Cur decreased FLT3 protein levels and total cell numbers in all the cell lines in a dose-dependent manner. In summary, this study exhibits a novel report of Dox–Cur co-treatment in both enhancing cytotoxicity of Dox and inhibiting cell proliferation via FLT3 protein expression in leukemia stem cells and leukemic cells. This is the option of leukemia treatment with reducing side effects of chemotherapeutic drugs to leukemia patients.     

Eight Indole Alkaloids from the Roots of Maerua siamensis and Their Nitric Oxide Inhibitory Effects

Maerua siamensis (Capparaceae) roots are used for treating pain and inflammation in traditional Thai medicine. Eight new indole alkaloids, named maeruanitriles A and B, maeroximes A–C, and maeruabisindoles A–C, were isolated from them. Spectroscopic methods and computational analysis were applied to determine the structure of the isolated compounds. Maeroximes A–C possesses an unusual O-methyloxime moiety. The bisindole alkaloid maeruabisindoles A and B possess a rare azete ring, whereas maeruabisindole C is the first indolo[3,2-b]carbazole derivative found in this plant family. Five compounds [maeruanitriles A and B, maeroxime C, maeruabisindoles B, and C] displayed anti-inflammatory activity by inhibiting nitric oxide (NO) production in the lipopolysaccharide-induced RAW 264.7 cells. Maeruabisindole B was the most active inhibitor of NO production, with an IC₅₀ of 31.1 ± 1.8 μM compared to indomethacin (IC₅₀ = 150.0 ± 16.0 μM) as the positive control.     

Simultaneous HPLC Analysis of Catecholamines and Indoleamines in Mouse Brain Tissue Following Acetate Extraction and Treatment with Ascorbate Oxidase

Abstract

A refined HPLC Method for the determination of monoamine levels in six brain regions is presented. Analyses are made for the olfactory tubercles, prefrontal cortex, septum, striatum, amygdala and hypothalamus of adult male ICR mice. This system permits the simultaneous analysis of norepinephrine, dopamine, serotonin and their major metabolites during a single run of approximately twenty-five minutes without prior clean-up of samples.     

Modification of Polyimide in Barrier Discharge Air-Plasmas: Chemical and Morphological Effects

Chemical and physical modifications of polyimide (PI) surfaces caused by an air plasma have been studied. The plasma-induced surface changes of PI were investigated by using a local dielectric barrier discharge (DBD) in air at atmospheric pressure and room temperature as a function of the plasma exposure time and plasma power, while the excitation frequency was kept constant at about 130 kHz. The first results obtained in this work suggest that DBDs operating in air at atmospheric pressure can be an efficient alternative plasma source for surface treatment of polymers: a short time air plasma treatment of few seconds leads to chemical and physical changes including the rise of wettability, surface oxidation, and enhancement of surface roughness. Therefore, this simple kind of dry surface treatment seems to be an effective, low cost method for production of well-adhering subsequent layers such as metal films, paints, glues, etc. on DBD pretreated polymers.     

Preparation of Active Peptides from Camellia vietnamensis and Their Metabolic Effects in Alcohol-Induced Liver Injury Cells

Liver damage seriously affects human health. Over 35% of cases of acute liver damage are caused by alcohol damage. Thus, finding drugs that can inhibit and effectively treat this disease is necessary. This article mainly focuses on the effect of the metabolome physical activity of active peptides in Camellia vietnamensis active peptide (CMAP) and improving liver protection. DEAE Sepharose FF ion-exchange column chromatography was used in separating and purifying crude peptides from Camellia vietnamensis Two components, A1 and A2, were obtained, and the most active A1 was selected. Sephadex G-100 gel column chromatography was used in A1 separation and purification. Three components, Al-1, Al-2, and Al-3, were obtained. Through antioxidant activity in vitro as an index of inspection, the relatively active component A1-2 was removed. Reverse-phase high-performance liquid chromatography showed that the purity of component A1-2 was 93.45%. The extracted CMAPs acted on alcoholic liver injury cells. Metabolomics studies revealed that the up-regulated metabolites were ribothymidine and xanthine; the down-regulated metabolites were hydroxyphenyllactic acid, creatinine, stearoylcarnitine, and inosine. This study provides an effective theoretical support for subsequent research.