Determination of the major constituents in fruit of Arctium lappa L. by matrix solid-phase dispersion extraction coupled with HPLC separation and fluorescence detection

The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r > 0.9998) in the concentration range of 0.010–5.0 μg mL⁻¹ for arctiin and 0.025–7.5 μg mL⁻¹ for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.     

HFRS患者特异性IgA,IgE抗体及其免疫复合物测定

为进一步研究ＨＦＲＳ免疫损伤机制,用ＥＬＩＳＡ法同步测定了１０８例不同临床型、不同病日、病期ＨＦＲＳ患者血清中特异性ＩｇＡ、ＩｇＥ抗体以及ＨＦＲＳ病毒特异性ＩｇＡ、ＩｇＥ型ＣＩＣ的水平及检出率。发现ＨＦＲＳＩｇＡ型抗体在轻型病例高于中、重型病例;ＨＦＲＳＩｇＥ型抗体及ＩｇＥ型ＣＩＣ在重型病例高于中、轻型病例。上述差异在病程早期（发热、休克少尿期,或是３～８病日）尤为突出。ＩｇＡ型ＣＩＣ则未见到上述差异。     

Pardaxin-induced apoptosis enhances antitumor activity in HeLa cells

Pardaxin, a pore-forming antimicrobial peptide that encodes 33 amino acids was isolated from the Red Sea Moses sole, Pardachirus mamoratus. In this study, we investigated its antitumor activity in human fibrosarcoma (HT-1080) cells and epithelial carcinoma (HeLa) cells. In vitro results showed that the synthetic pardaxin peptide had antitumor activity in these two types of cancer cells and that 15 μg/ml pardaxin did not lyse human red blood cells. Moreover, this synthetic pardaxin inhibited the proliferation of HT1080 cells in a dose-dependent manner and induced programmed cell death in HeLa cells. DNA fragmentation and increases in the subG1 phase and caspase 8 activities suggest that pardaxin caused HeLa cell death by inducing apoptosis, but had a different mechanism in HT1080 cells.     

聚乙二醇化学修饰的聚乙烯亚胺衍生物的制备及其细胞毒性研究

目的:合成聚乙二醇(PEG)化的聚乙烯亚胺衍生物(PEI-Et)基因输送载体PET 1和PET 2,并考察两个载体材料在He La细胞、MCF-7细胞中的细胞毒性及在He La细胞中的转染效率。方法:将亚乙基二氯甲酸酯与PEI 800 Da交联制备成交联PEI衍生物PEI-Et,进一步将PEI-Et与聚乙二醇(PEG)以不同摩尔比例(1:1,2:1)交联连接,得到PEG化的PET 1和PET 2。采用MTT法检测PEI-Et、PET 1、PET 2对He La细胞、MCF-7细胞的细胞毒性。检测单位质量的荧光强度测定转染效率。结果:PET的细胞毒性随浓度增大而增大,在同一浓度下PET的细胞毒性小于PEI 25 KDa(P0.01);并且与DNA复合后,复合物细胞毒性随质量比的增高而增大,在同一质量比下PET的细胞毒性小于PEI 25 KDa(P0.01),特别是PET 1。并且最佳比例时,PET 1的转染活性最高。结论:作为非病毒基因载体,PET 1具有高的转染效率及低的细胞毒性。     

Kelsoenethiol and dikelsoenyl ether,two unique kelsoane-type sesquiterpenes,from the Formosan soft coral Nephthea erecta

Two new kelsoane-type sesquiterpenes, namely kelsoenethiol (1) and dikelsoenyl ether (2), were obtained from the Formosan soft coral Nephthea erecta. Their structures were elucidated through extensive spectroscopic analyses, ESI orbitrap mass and quantum chemical calculations (QCC). The cytotoxicity against A-459 (human lung carcinoma), P-388 (mouse lymphocytic leukemia), and HT-29 (human colon adenocarcinoma) cancer cell lines of 1 and 2 was evaluated in vitro. Compound 1 showed cytotoxicity against P-388 and HT-29 cells with ED_50s of 1.3 and 1.8 μg/mL, respectively.     

Evaluation of ARG protein expression in mature B cell lymphomas compared to non-neoplastic reactive lymph node

The participation of Abl-Related Gene (ARG) is demonstrated in pathogenesis of different human malignancies. However there is no conclusive evidence on ARG expression level in mature B cell lymphomas. In this study we evaluated ARG protein expression in Follicular Lymphoma (FL), Burkitt’s Lymphoma (BL) and Diffused Large B Cell Lymphoma (DLBCL) in comparison with non-neoplastic lymph nodes. Semi-quantitative fluorescent ImmunoHistoChemistry was applied on 14, 7 and 4 patients with DLBCL, FL and BL respectively, adding to 4 normal and 4 reactive lymph nodes. The mean ratio of ARG/GAPDH expression was significantly different (p < 0.00) between lymphomas and control samples, with DLBCL having the highest ARG expression amongst all. Over expression of ARG was seen in FL and BL, with FL expressing statistically more ARG than BL. Moreover, the ARG/GAPDH expression ratio increased from DLBCL stage I towards stage VI, all showing significantly more ARG expression than FL and BL (in all cases p < 0.00).     

Tumor cytotoxicity of 5,6-dimethyl-1,10-phenanthroline and its corresponding gold(III) complex

A gold(III) complex possessing 5,6-dimethyl-1,10-phenanthroline (5,6DMP) was synthesized and fully characterized using standard spectroscopic techniques, as well as X-ray crystallography and elemental analysis. The complex [(5,6DMP)AuCl₂][BF₄] (2) was found to possess a distorted square planar geometry about the gold(III) center, commonplace for d⁸ Au(III) cations possessing sterically un-hindered polypyridyl ligands. Compound 2 was evaluated for its potential use as an anticancer therapeutic. It was determined that the complex is stable in phosphate buffer over a 24-hour period, thought it does undergo rapid reduction in the presence of equimolar amounts of reduced glutathione (GSH) and ascorbic acid. The DNA binding and in vitro tumor cytotoxicity of the title compound 2 were also determined. It was found to undergo weak and reversible binding to calf thymus DNA, and was more cytotoxic towards a panel of human cancer cell lines than the commonly used chemotherapy agent cisplatin. Cytotoxicity experiments with the free 5,6DMP ligand indicate that the ligand has IC₅₀ values that are slightly lower than those observed for the gold complex (2), and coupled with the fact that the ligand appears to be released from the gold(III) metal center in reducing environments, this suggests the ligand itself may play an important role in the antitumor activity of the parent gold complex.     

Functional properties of FC-2.15, a monoclonal antibody that mediates human complement cytotoxicity against breast cancer cells

FC-2.15 is a murine IgM monoclonal antibody (mAb) that recognizes a cell-surface antigen (Ag2.15) expressed in most tumor-proliferating cells of human breast carcinomas and other neoplasias. In this study the cytotoxic ability of mAb FC-2.15, its cell-surface binding properties and endocytosis in Ag2.15-expressing (Ag2.15⁺) cells were investigated. A⁵¹Cr-release assay was used to test the FC-2.15-mediated cytotoxicity. When human serum was used as source of complement, FC-2.15 exerted a strong cytotoxic effect against human Ag2.15⁺ cells such as MCF-7 (breast cancer cell line), primary breast carcinoma cells, polymorphonuclear leukocytes and chronic myeloid leukemia cells. The mAb concentration range was 1–50 g/ml. Cytotoxicity was completely abolished when complement was inactivated. Only 3.8±2.9% of MCF-7 cells survived the treatment with FC-2.15 in the presence of human serum. A flow-cytometry assay was performed to study the Ag2.15 expression of the surviving cells and they were found to be Ag2.15^–. FC-2.15 did not mediate antibody-dependent cell cytotoxicity when different effector cells were used. Scatchard analysis with¹²⁵I-FC-2.15 on MCF-7 cells demonstrated an affinity constant of 6.9×10⁷ M^–1 and 2.8×10⁶ antigenic sites/cell.¹²⁵I-FC-2.15 was internalized to cytoplasmic vesicles reaching a maximum of 27% after 6 h incubation, followed by the release of labeled degradation products to the supernatant. FC-2.15 appears to exert its cytotoxic effect mainly in the presence of human complement, it reacts with intermediate affinity with a high-density surface antigen, and it is slowly internalized by Ag2.15⁺ cells.     

Pt(II) complexes with (N,N′) or (C,N,E)− (E = N,S) ligands: Cytotoxic studies,effect on DNA tertiary structure and structure–activity relationships

The cytotoxic activity of two series of platinum(II) complexes containing the polyfunctional imines R¹–CHN–R² [R¹ = phenyl or ferrocenyl unit and R² = (CH₂)_n–CH₂–NMe₂ where n = 1 or 2) (1 and 2) or C₆H₄-2-SMe (3)] acting as a bidentate (N,N′) (4–7) or terdentate [C(phenyl or ferrocenyl),N,N′]^? (8–10) or [C(ferrocenyl),N,S]^? ligand (11) in front of A549 lung, MDA-MB231 breast and HCT116 colon human adenocarcinoma cell lines is reported. The results reveal that most of the platinum(II) complexes are active against the three assayed lines and compounds 6, 7 and the platinacycles 10 and 11 exhibit a remarkable antiproliferative activity, even greater than cisplatin itself, in the cisplatin resistant HCT116 human cancer cell line. Electrophoretic DNA migration studies showed that most of them modify the DNA tertiary structure in a similar way as the reference cisplatin. Solution studies of a selection of the most relevant complexes have also been performed in order to test: (a) their stability in the aqueous biological medium and/or the formation of biologically active species and (b) their proclivity to react with 9-methylguanine (9-MeG), as a model nucleobase. Computational studies at DFT level have also been performed in order to explain the different solution behaviour of the complexes and their proclivity to react with the nucleobase.     

检测肾综合征出血热抗原特异性循环免疫复合物ELISA方法的建立

检测肾综合征出血热抗原特异性循环免疫复合物ＥＬＩＳＡ方法的建立张东海,孙辉,高峰（山东省淄博第二卫生学校,淄博２５５０１５）关键词肾综合征出血热病毒,循坏免疫复合物,特异性,ＥＬＩＳＡ肾综合征出血热（ＨＦＲＳ）发病机理有多种学说,其中在体液免疫学说中...     

果蝇亚群中六个种的求爱歌的研究：对ipi作用的探究

据认为果蝇求爱歌的脉冲间隔(ipi)在种特异性配偶认识系统(SMRS)中起着重要的作用。果蝇D.takahashii亚群中各物种求爱歌的ipi值和种间亲缘关系的相关性如何?迄今尚未见报道。我们测定了此亚群中能饲养存活的6个种的ipi,并用O’Farrell双向电泳方法及杂交实验研究了它们之间的亲缘关系。在近缘种D.takahashii和D.lutescens之间,D.takahashii和D.paralutea之间,D.lutescens和D.paralutea之间,其求爱歌的ipi值,不论平均数或众数都相差甚远。而D.paralutea和D.prostipennis是2个亲缘关系较远的种,两者不能杂交,但其ipi的平均数在统计学上无差异,众数亦很相近。本实验可导出这样的见解:ipi值在不同果蝇种群或亚群的SMRS中的作用可能并不相同。在D.takahashii亚群中ipi值对于杂交成败似乎不起支配性作用。     

Kinetics and mechanism of the complex formation of [Pd(NNN)Cl] with pyridines in methanol: synthesis and crystal structure of [Pd(terpy)(py)](ClO4)2

The kinetics of the complex-formation reactions between monofunctional palladium(II) complexes [Pd(NNN)Cl]⁺, where NNN is 2,2^′:6^′,2″-terpyridine (terpy), diethylenetriamine (dien) or bis(2-pyridylmethyl)amine (bpma), with pyridine, 4-methylpyridine, 4-acetylpyridine, 4-cyanopyridine and 4-aminopyridine, have been studied in methanol at 25 °C using stopped-flow spectrophotometry. The highest reactivity was observed for the [Pd(terpy)Cl]⁺ complex, whereas 4-aminopyridine is the strongest nucleophile. The results, compared with those previously published on the [Pt(NNN)Cl]⁺ complexes, are discussed in terms of reactivity and discrimination ability of the reaction centre. The crystal structure of [Pd(terpy)(py)](ClO₄)₂ has been determined by X-ray diffraction. Crystals are triclinic, space group , and consist of distorted square planar [Pd(terpy)(py)]²⁺ cations and perchlorate anions. The Pd-N bond length to the central atom of terpy ligand is well below 2.0 Å and significantly shorter than any of the other M-N distances. The pyridine plane forms a dihedral angle of 61.9(2)° with the coordination N4 donors.     

Synthesis and biological evaluation of arctigenin ester and ether derivatives as activators of AMPK

A series of new arctigenin and 9-deoxy-arctigenin derivatives bearing different ester and ether side chains at the phenolic hydroxyl positions are designed, synthesized, and evaluated for activating AMPK potency in L6 myoblasts. Initial biological evaluation indicates that some alkyl ester and phenethyl ether arctigenin derivatives display potential activities in AMPK phosphorylation improvement. Further structure–activity relationship analysis shows that arctigenin ester derivatives 3a, 3h and 9-deoxy-arctigenin phenethyl ether derivatives 6a, 6c, 6d activate AMPK more potently than arctigenin. Moreover, the 2-(3,4-dimethoxyphenyl)ethyl ether moiety of 6c has been demonstrated as a potential functional group to improve the effect of AMPK phosphorylation. The structural optimization of arctigenin leads to the identification of 6c as a promising lead compound that exhibits excellent activity in AMPK activation.     

Upregulation of PBR mRNA expression in human neuroblastoma cells by flavonoids

To investigate the putative mediation of peripheral benzodiazepine receptor (PBR) in the cytotoxicity of flavonoids, in this study, modulatory effects of several flavonoids on the lipid peroxide (LPO) production and PBR mRNA expression of human neuroblastoma cells were observed. Elevated levels of peroxidated products in cancer cells may activate pro-apoptotic and anti-proliferative signaling pathways. Treatment of 10(-6) M 4'-chlorodiazepam and PK 11195 ligands of the PBR for 6 days enhanced the generation of LPO of the human neuroblastoma cells. Several flavonoids, well-known cytotoxic substances, potentiated the enhancement of LPO production by PBR ligands. Treatment of 10(-6) M flavonoids for 6 days elevated the expression of PBR mRNA in cells. These findings indicate that the potential of flavonoids to induce apoptosis in cancer cells is strongly associated with their PBR-inducing properties, thereby providing a new mechanism by which polyphenolic compounds may exert their cancer-preventive and anti-neoplastic effects.     

Molecular modeling,synthesis, antibacterial and cytotoxicity evaluation of sulfonamide derivatives of benzimidazole,indazole, benzothiazole and thiazole

A new series of heterocyclic molecules bearing sulfonamide linkage has been synthesized and screened for antibacterial activity. During antibacterial screening using broath dilution method, molecules were found to be highly active (MIC value 50–3.1?µg/mL) against different human pathogens, namely B. cerus, S. aureus, E. coli and P. aeruginosa, and most effective against E. coli. A great synergistic effect was observed during determination of FIC where molecules were used in combination with reference drugs chloramphenicol and sulfamethoxazole. The MIC value of the combination – varying concentration of test compounds and ½ MIC of reference drugs or varying concentration of reference drugs and ½ MIC of test compounds, was reduced up to ¹/₄ or ¹/₃₂ of the original value, indicating thereby the combination was 4–32 times more potent than the test molecule. The molecules also showed low degree of cytotoxicity against PBM, CEM and VERO cell lines. The results positively indicated towards the development of lead antibacterials using the combination approach.     

Comparative study of the influence of the metal centres: Fe(III), Cu(II) and Zn(II), on the ring opening and oxidative dehydrogenation reactions occurring in a coordinated imidazolidine ligand

Condensation of 2-pyridinecarboxaldehyde and 1,9-bis-(2′-pyridyl)-2,5,8-triazanonane, L¹, yields 1-[3-aza-4-(2-pyridyl)butyl]-2-(2-pyridyl)-3-[(2-pyridyl)methyl]imidazolidine, L², as proven by NMR solution spectra. When L² is reacted with Fe(III) in different alcohols, an imidazolidine ring opening and an oxidative dehydrogenation reaction occur resulting in new complexes of the type: [Fe^IIL^n′]²⁺. Compound 1 with a coordinated L^3′ ligand was obtained in n-propanol as a solvent. Compounds 2, 3 and 4 were obtained with L^4′, L^5′ and L^6′ when iso-propanol, n-butanol and iso-butanol were used as solvent, respectively. The structures for 1, 2, 3 and 4 were determined by NMR solution spectra and additionally by X-ray crystallography in the case of the n-butoxy derivative 3. When Cu(II) was used, the hexadentate ligand L² undergoes also an imidazolidine ring opening reaction on complex formation, however, now generating the well-known pentadentate ligand L¹ that is coordinated to the metal ion, 7. Evidence is again provided by the corresponding X-ray structure. With Zn(II) the initial structure of L² is maintained and in this case L² functions as a tetradentate, 5, or bis-tridentate ligand, 6, depending on whether the stoichiometric ratio M:L was 1:1 or 2:1, respectively. This has been proven by a solid-state X-ray structure analysis as well as by NMR solution spectra. The ring opening reaction in the presence of Fe(III) can be explained as a result of a higher Lewis acidity of this metal centre, which decreases the electronic density on the nitrogen atom of the imidazolidinic cycle, thus weakening the nitrogen-carbon bond, favouring the nucleophilic attack on the carbon atom by alcohols and producing a more stable hexacoordinated species. Electrochemical evidence is provided in order to support this reaction mechanism.