Fused Polypeptide with DEF Induces Apoptosis of Lung Adenocarcinoma Cells

Liang, Ai-Ling,Zhang, Ting-Ting,Zhou, Ning,Huang, Di-Nan,Liu, Xin-Guang,Liu, Yong-Jun,Tu, Zhi-Guang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.12

To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT-DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.

Alkaloids from Beach Spider Lily (Hymenocallis littoralis) Induce Apoptosis of HepG-2 Cells by the Fas-signaling Pathway

Ji, Yu-Bin,Chen, Ning,Zhu, Hong-Wei,Ling, Na,Li, Wen-Lan,Song, Dong-Xue,Gao, Shi-Yong,Zhang, Wang-Cheng,Ma, Nan-Nan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose ($0.8{\mu}g/ml$) significantly inhibiting proliferation. The non-tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

Environmental Microbiology and Engineering : Microbial Community Dynamics in Batch High-Solid Anaerobic Digestion of Food Waste Under Mesophilic Conditions

( Jing Yi ),( Bin Dong ),( Yong Gang Xue ),( Ning Li ),( Peng Gao ),( Yu Xin Zhao ),( Ling Ling Dai ),( Xiao Hu Dai ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2

Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g VSremoval. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

In vitro Chemosensitivity in Breast Cancer Using ATP-tumor Chemosensitivity Assay

Chun-Jian Qi,Yong-Ling Ning,Yu-Lan Zhu,Hai-Yan Min,Heng Ye,Ke-Qing Qian 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.12

Chemotherapy for breast cancer is given on the basis of empirical information from clinical trials, an approach which falls to take into account the known heterogeneity of chemosensitivity between patients. This study aimed to demonstrate the degree of heterogeneity of chemosensitivity in breast cancers. In this study, we examined the heterogeneity of chemosensitivity in breast cancer specimens (n = 50) using an ex vivo ATP-tumor chemosensitivity assay (ATP-TCA). Assay evaluability was 92% in surgical biopsies or pleural aspirates. A variety of chemosensitivity agents were tested. We found that the most active single agent tested was paclitaxel, to which 65.9% of samples were sensitive. Combinations of agents also showed more strong sensitivity cases. The Adriamycin+5-FU demonstrated a strong sensitivity in 23 of 43 (52.3%) of samples. Adriamycin+paclitaxel was more effective, with strong sensitivity in 37 of 43 cases tested (86.0%). There was a marked heterogeneity of chemosensitivity in breast cancer. Chemosensitivity testing may provide a practical method of testing new regimens before clinical trials in breast cancer patients.

Microstructure and tribological properties of ion beam‑modified GO‑reinforced copper matrix composites

Zi‑liang Liu,Yong Li,Xiao Guo,Jun‑jie Tao,Ji‑ning Huang,Ling‑lin Fang 한국탄소학회 2023 Carbon Letters Vol.33 No.6

The flaw of low dispersibility in the metal matrix brought on by graphene's full crystal structure can be improved by the application of ion beam radiation to the surface of the material. Copper atoms are uniformly dispersed on the modified graphene oxide ( GOM) surface after being irradiated to a copper ion beam, and during the sputtering modification, the valence state of copper is changed, resulting in the formation of a new CuO phase on the graphene oxide (GO) surface. Therefore, after copper ion beam irradiation of graphene, the interfacial adhesion between GOM and copper matrix is enhanced, and the wear resistance is significantly improved. When the GOM content is low, it can withstand most of the load during the friction and wear test, which reduces the wear of the copper matrix and the occurrence of fatigue cracks at the interface of the composite material.

In Vivo Protein Transduction: Delivery of PEP-1-SOD1 Fusion Protein into Myocardium Efficiently Protects against Ischemic Insult

You-en Zhang,Jia-ning Wang,Jun-ming Tang,Ling-yun Guo,Jian-ye Yang,Yong-zhang Huang,Yan Tan,Shou-zhi Fu,Xia Kong,Fei Zheng 한국분자세포생물학회 2009 Molecules and cells Vol.27 No.2

Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1-SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

Highly Selective Production of Compound K from Ginsenoside Rd by Hydrolyzing Glucose at C-3 Glycoside Using β-Glucosidase of Bifidobacterium breve ATCC 15700

( Ru Zhang ),( Xue-mei Huang ),( Hui-juan Yan ),( Xin-yi Liu ),( Qi Zhou ),( Zhi-yong Luo ),( Xiao-ning Tan ),( Bian-ling Zhang ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.3

To investigate a novel β-glucosidase from Bifidobacterium breve ATCC 15700 (BbBgl) to produce compound K (CK) via ginsenoside F2 by highly selective and efficient hydrolysis of the C-3 glycoside from ginsenoside Rd, the BbBgl gene was cloned and expressed in E. coli BL21. The recombinant BbBgl was purified by Ni-NTA magnetic beads to obtain an enzyme with specific activity of 37 U/mg protein using pNP-Glc as substrate. The enzyme activity was optimized at pH 5.0, 35°C, 2 or 6 U/ml, and its activity was enhanced by Mn²⁺ significantly. Under the optimal conditions, the half-life of the BbBgl is 180 h, much longer than the characterized β-glycosidases, and the Km and V_max values are 2.7 mM and 39.8 μmol/mg/min for ginsenoside Rd. Moreover, the enzyme exhibits strong tolerance against high substrate concentration (up to 40 g/l ginsenoside Rd) with a molar biotransformation rate of 96% within 12 h. The good enzymatic properties and gram-scale conversion capacity of BbBgl provide an attractive method for large-scale production of rare ginsenoside CK using a single enzyme or a combination of enzymes.