Preparative isolation and purification of anti‐tumor agent ansamitocin P‐3 from fermentation broth of Actinosynnema pretiosum using high‐performance counter‐current chromatography

Ansamitocin P‐3 is a potent anti‐tumor maytansinoid found in Actinosynnema pretiosum. However, due to the complexity of the fermentation broth of Actinomycete, how to effectively separate ansamitocin P‐3 is still a challenge. In this study, both analytical and preparative high‐performance counter‐current chromatography were successfully used to separate and purify ansamitocin P‐3 from fermentation broth. A total of 28.8 mg ansamitocin P‐3 with purity of 98.4% was separated from 160 mg crude sample of fermentation broth in less than 80 min with the two‐phase solvent system of hexane–ethyl acetate–methanol–water (0.6:1:0.6:1, v/v/v/v). The purity and structural identification were determined by HPLC, ¹H NMR, ¹³C NMR and mass spectroscopy.     

Sonodynamically induced anti-tumor effect with protoporphyrin IX on hepatoma-22 solid tumor

Objective

The purpose of this study was to evaluate sonodynamically induced anti-tumor effect of protoporphyrin IX (PPIX) in mice bearing hepatoma-22 (H-22) solid tumors, and the possible in vivo cell damage mechanism was also investigated.

Methods

The pharmacokinetics of PPIX was analyzed in plasma, skin, muscle and tumor of H-22 bearing mice. Tumors were irradiated with ultrasound (1.43 MHz, I_SATA 3 W/cm², 3 min) for three times at 8, 12 and 24 h after 5.0 mg/kg PPIX administration, respectively. The anti-tumor effects of sonodynamic therapy (SDT) were estimated by the tumor inhibition ratio (volume and weight). The bio-effects of SDT were evaluated by hematoxylin and eosin (H&E) staining, transmission electron microscope (TEM) observation, lipid peroxidation (LPO) measurement and anti-oxidative enzymes (glutathione peroxidase (GSH-PX), catalase (CAT) and superoxide dismutase (SOD)) assay.

Results

A significant anti-tumor effect was obtained by PPIX-mediated sonodynamic therapy (PPIX-SDT). At the fifteenth day after PPIX-SDT, the tumor growth and tumor weight inhibition ratios were 53.84% and 45.86%, respectively. In addition, the structure of tumor tissues and the anti-oxidative enzymes were obviously destroyed after SDT treatment.

Conclusions

A biochemical mechanism was involved in PPIX-SDT in vivo, and the free radicals produced by the synergistic treatment destroying the anti-oxidative system of tumor cells in vivo may play an important role in this action. Also, the thermal effect could not be excluded in inducing damage of cellular structures, like membrane disruption and chromatin condensation under current evaluation in this paper.     

Fluctuations induced extinction and stochastic resonance effect in a model of tumor growth with periodic treatment

We investigate a stochastic model of tumor growth derived from the catalytic Michaelis-Menten reaction with positional and environmental fluctuations under subthreshold periodic treatment. Firstly, the influences of environmental fluctuations on the treatable stage are analyzed numerically. Applying the standard theory of stochastic resonance derived from the two-state approach, we derive the signal-to-noise ratio (SNR) analytically, which is used to measure the stochastic resonance phenomenon. It is found that the weak environmental fluctuations could induce the extinction of tumor cells in the subthreshold periodic treatment. The positional stability is better in favor of the treatment of the tumor cells. Besides, the appropriate and feasible treatment intensity and the treatment cycle should be highlighted considered in the treatment of tumor cells.     

Theoretical Analysis of the Biological Thermal Effect of Millimeter Waves in Layered-Dielectric-Slabs

The theory is presented for one method of determining the biological thermal effect of millimeter waves in microwave radiometry. It has been studied theoretically that millimeter waves propagation and absorption in a human body. The model is a plane straticulate homogeneous slab of tissues under the irradiance of normal incidence plane wave. It has been discussed by obtaining the electromagnetic field, absorbent power, specific absorption rate, temperature field and their distributions in the human trunk model. Also, the principle of thermal therapeutics of millimeter waves to cancer has been discussed preliminarily.     

Radiobiological response of human hepatoma and normal liver cells exposed to carbon ions generated by Heavy Ion Research Facility in Lanzhou

Human hepatoma and normal liver cells were irradiated with ¹²C⁶⁺ ion beams (LET = 96.05 keV/μm) and γ-rays at Heavy Ion Research Facility in Lanzhou (HIRFL). The chromatid breaks and break types were detected using the premature chromosome condensation technique. Our experimental results showed that chromatid breaks seem to have a good relation with ¹²C⁶⁺ absorbed dose and ¹²C⁶⁺ are more effective to induce chromatid breaks as compared to the γ-rays. For ¹²C⁶⁺ ion irradiation the major break was isochromatid break, while chromatid breaks were dominant for γ-ray irradiation. We also observed that the Relative Biology Effectiveness (RBE) of ¹²C⁶⁺ ion is about 2.5 times higher than that of γ-rays.     

抑制性消减杂交方法的建立

为建立研究基因表达差异的抑制性消减杂交（SSH）方法，以未加诱导剂处理的结肠癌LoVo细胞提取的mRNA为模板合成的cDNA作为驱赶子（driver)，联合使用10.0μmol/LATRA和0.1μmol/L1,25-(OH)2D3诱导2d的LoVo细胞提取的mRNA为模板合成的cDNA作为测试子（tester)进行SSH实验，结果为：消减产物与未消减样品（对照）在琼脂糖凝胶电泳图谱上明显不同，前者可见一些扩增条带，其大小范围在100-1000bp之间，本实验说明SSH方法对于识别差异表达的基因具有高效性。     

Far‐Red and Near‐IR AIE‐Active Fluorescent Organic Nanoprobes with Enhanced Tumor‐Targeting Efficacy: Shape‐Specific Effects

The rational design of high‐performance fluorescent materials for cancer targeting in vivo is still challenging. A unique molecular design strategy is presented that involves tailoring aggregation‐induced emission (AIE)‐active organic molecules to realize preferable far‐red and NIR fluorescence, well‐controlled morphology (from rod‐like to spherical), and also tumor‐targeted bioimaging. The shape‐tailored organic quinoline–malononitrile (QM) nanoprobes are biocompatible and highly desirable for cell‐tracking applications. Impressively, the spherical shape of QM‐5 nanoaggregates exhibits excellent tumor‐targeted bioimaging performance after intravenously injection into mice, but not the rod‐like aggregates of QM‐2.     

Fluorescent In Situ Targeting Probes for Rapid Imaging of Ovarian‐Cancer‐Specific γ‐Glutamyltranspeptidase

γ‐Glutamyltranspeptidase (GGT) is a tumor biomarker that selectively catalyzes the cleavage of glutamate overexpressed on the plasma membrane of tumor cells. Here, we developed two novel fluorescent in situ targeting (FIST) probes that specifically target GGT in tumor cells, which comprise 1) a GGT‐specific substrate unit (GSH), and 2) a boron–dipyrromethene (BODIPY) moiety for fluorescent signalling. In the presence of GGT, sulfur‐substituted BODIPY was converted to amino‐substituted BODIPY, resulting in dramatic fluorescence variations. By exploiting this enzyme‐triggered photophysical property, we employed these FIST probes to monitor the GGT activity in living cells, which showed remarkable differentiation between ovarian cancer cells and normal cells. These probes represent two first‐generation chemodosimeters featuring enzyme‐mediated rapid, irreversible aromatic hydrocarbon transfer between the sulfur and nitrogen atoms accompanied by switching of photophysical properties.     

Folic acid‐polyamine‐β‐cyclodextrin for targeted delivery of scutellarin to cancer cells

The efficient tumor targeting drug carrier was designed by bioconjugation of folic acid to β‐cyclodextrin through a polyamine cationic spacer. The characterization and inclusion complexation behavior of the inclusion complex of hydrophobic drug scutellarin with folic acid‐polyamine‐β‐cyclodextrin were investigated in both solution and solid state by means of phase‐solubility, nuclear magnetic resonance, X‐ray power diffraction, thermal gravimetric analysis, and scanning electron microscopy. Besides, the solubilization efficiency and antitumor activity of the inclusion complex were tested by saturated solution and MTT (Thiazole blue) method. Solubility and antitumor activity studies showed higher solubilizing ability and antitumor activity of the inclusion complex in comparison to free scutellarin. The folic acid‐polyamine‐β‐cyclodextrin that is presented may be promising active tumor‐targeting carrier candidates via folate mediation. Copyright © 2015 John Wiley & Sons, Ltd.