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      • KCI등재

        Soluble expression, purification and the role of C-terminal glycine residues in scorpion toxin BmK AGP-SYPU2

        ( Rong Zhang ),( Yong Cui ),( Xi Zhang ),( Zhuo Yang ),( Yong Shan Zhao ),( Yong Bo Song ),( Chun Fu Wu ),( Jing Hai Zhang ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.12

        The existence of glycine residues in long-chain scorpion toxins has been well documented. However, their role as analgesics has not been evaluated. To address this issue, we investigated the functional role of glycines in the C-terminal end of Chinese-scorpion toxin from Buthus martensii Karsch (BmK AGP-SYPU2) using site-directed mutagenesis and analgesic activity assays. Recombinant BmK AGP-SYPU2 and its mutants were efficiently expressed in E. coli and purified to homogeneity using immobilized metal ion affinity chromatography (IMAC) and cation exchange chromatography. The mouse-twisting test was used to detect the analgesic activity of BmK AGP-SYPU2 and its mutants. As a result, we identified glycines at the C-terminal end that, when altered, significantly affected analgesic activity. Also, Mut6566 was significantly decreased compared to BmK AGP-SYPU2. These data indicate that the glycines at the C-terminal end are important for the analgesic activity of BmK AGP-SYPU2. [BMB reports 2010; 43(12): 801-806]

      • KCI등재

        Studies on PEGylated Gold Nanoparticles Loaded with 5-Hydroxydecanoate for Chemo-Photothermal Therapy of Human Lung Adenocarcinomas In Vitro

        Fu-Yan Zhuge,CONG-WANG ZHANG,CHANG-CHUN ZENG,Han-Ping Liu 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2015 NANO Vol.10 No.7

        Gold nanoparticles (Au NPs) have promising applications in the fields of drug delivery and photothermal therapy. 5-hydroxydecanoate (5-HD) is a kind of highly selective mitochondrial ATP sensitive potassium (mitoKATP) channel blocker. In this research, firstly, 5-HD was studied whether it could induce human lung adenocarcinomas (A549) cells apoptosis; secondly, PEGylated gold nanoparticles loaded with 5-HD (Au NPs-PEG-5-HD) were prepared to develop a new chemotherapy and photothermal therapy in one system under the irradiation of green light-emitting diode (LED). Subsequently, in vitro cytotoxicity test was analyzed by cell counting kit-8 (CCK-8), the change of mitochondrial membrane potential (Δψm) was determined by immunofluorescence microscopy with R-123 fluorescence, and cell apoptosis and necrosis rate were detected by flow cytometry. The results of CCK8 revealed that the inhibition rates of A549 cells were all greatly increased when cells were treated with free 5-HD, free 5-HD +LED irradiation, Au NPs-PEG-5-HD and Au NPs-PEG-5-HD+LED irradiation, and Au NPs-PEG-5-HD had enhanced cell-killing effect compared with 5-HD, furthermore, the Au NPs-PEG-5-HD and LED irradiation played a very great synergy effect. The immunofluorescence microscopy data also exhibited a reduction of Δψm correspondingly. Flow cytometric analysis showed that the apoptosis rate of cells that were incubated with free 5-HD, 5-HD+LED irradiation, Au NPsPEG-5-HD or Au NPs–PEG–5-HD+LED irradiation significantly increased to about 9.2%, 10.7%, 18.3% or 12.4% and the percentage of necrosis cells increased to around 8.8%, 9.7%, 48.0% or 69.8%, respectively. In a word, all the results indicated that the 5-HD had shown the ability to induce A549 cells apoptosis as a chemotherapy agent, and its ability would be improved when 5-HD is loaded on PEGylated Au NPs, as well as Au NPs-PEG-5-HD exhibited significantly enhanced photothermal effects for treatment of lung adenocarcinomas.

      • KCI등재

        Analysis of dynamic behavior for truss cable structures

        Wen-Fu Zhang,Ying-Chun Liu,Jing Ji,Zhen-Chao Teng 국제구조공학회 2014 Steel and Composite Structures, An International J Vol.16 No.2

        Natural vibration of truss cable structures is analyzed based upon the general structural analysis software ANSYS, energy variational method and Rayleigh method, the calculated results of three methods are compared, from which the characteristics of free-vibration are obtained. Moreover, vertical seismic response analysis of truss cable structures is carried out via time-history method. Introducing three natural earthquake waves calculated the results including time-history curve of vertical maximal displacement, time-history curve of maximal internal force. Variation curve of maximal displacement of node along span, and variation curve of maximal internal force of member along span are presented. The results show the formulas of frequencies for truss cable structures obtained by energy variational method are of high accuracy. Furthermore, the maximal displacement and the maximal internal force occur near the 1/5 span point. These provide convenient and simple design method for practical engineering.

      • KCI등재

        Definitive extended field intensity-modulated radiotherapy and concurrent cisplatin chemosensitization in the treatment of IB2-IIIB cervical cancer

        Guangyu Zhang,Fangfang He,Chunli Fu,Youzhong Zhang,Qiuan Yang,Jianbo Wang,Yufeng Cheng 대한부인종양학회 2014 Journal of Gynecologic Oncology Vol.25 No.1

        Objective: To assess the toxicity of delivering extended field intensity-modulated radiotherapy (EF-IMRT) and concurrent cisplatin chemotherapy for locally advanced cervical carcinoma. Methods: Forty-five patients who underwent EF-IMRT and concurrent cisplatin chemotherapy for the treatment of stage IB2 to IIIB cervical cancer were retrospectively reviewed. The clinical target volume included all areas of gross and potentially microscopic disease and regional lymph node regions. All patients underwent high-dose-rate brachytherapy. The acute and late toxicity were scored using the Common Terminology Criteria for Adverse Events and the Radiation Therapy Oncology Group late radiation morbidity scoring criteria, respectively. Results: The median follow-up was 28 months (range, 5 to 62 months). Forty-two patients had a complete response, and three had a persistent disease. Of those 42 patients, 15 patients (35.7%) had recurrence. The regions of recurrence were in-field in 2 patients and out-field in 13 patients. Acute grade ≥3 gastrointestinal, genitourinary and hematologic toxicity occurred in 3, 1, and 9 patients, respectively. Three patients (6.7%) suffered from late grade 3 toxicities. Seven patients experienced ovarian transposition, 5 of those patients (71%) maintained ovarian function. Thirty-eight patients (84.4%) were alive at the last follow-up. Conclusion: Concurrent cisplatin chemotherapy with EF-IMRT was safe. The acute and late toxicities are acceptable. EF-IMRT provides an opportunity to preserve endocrine function for patients with ovarian transposition.

      • Genomic Screening for Targets Regulated by Berberine in Breast Cancer Cells

        Wen, Chun-Jie,Wu, Lan-Xiang,Fu, Li-Juan,Yu, Jing,Zhang, Yi-Wen,Zhang, Xue,Zhou, Hong-Hao Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

        Berberine, a common isoquinoline alkaloid, has been shown to possess anti-cancer activities. However, the underlying molecular mechanisms are still not completely understood. In the current study, we investigated the effects of berberine on cell growth, colony formation, cell cycle distribution, and whether it improved the anticancer efficiency of cisplatin and doxorubicin in human breast cancer estrogen receptor positive (ER+) MCF-7 cells and estrogen receptor negative (ER-) MDA-MB-231 cells. Notably, berberine treatment significantly inhibited cell growth and colony formation in the two cell lines, berberine in combination with cisplatin exerting synergistic growth inhibitory effects. Accompanied by decreased growth, berberine induced G1 phase arrest in MCF-7 but not MDA-MB-231 cells. To provide a more detailed understanding of the mechanisms of action of berberine, we performed genome-wide expression profiling of berberine-treated cells using cDNA microarrays. This revealed that there were 3,397 and 2,706 genes regulated by berberine in MCF-7 and MDA-MB-231 cells, respectively. Fene oncology (GO) analysis identified that many of the target genes were involved in regulation of the cell cycle, cell migration, apoptosis, and drug responses. To confirm the microarray data, qPCR analysis was conducted for 10 selected genes based on previously reported associations with breast cancer and GO analysis. In conclusion, berberine exhibits inhibitory effects on breast cancer cells proliferation, which is likely mediated by alteration of gene expression profiles.

      • Preferential Induction of CYP1A1 over CYP1B1 in Human Breast Cancer MCF-7 Cells after Exposure to Berberine

        Wen, Chun-Jie,Wu, Lan-Xiang,Fu, Li-Juan,Shen, Dong-Ya,Zhang, Xue,Zhang, Yi-Wen,Yu, Jing,Zhou, Hong-Hao Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

        Estrogens are considered the major breast cancer risk factor, and the carcinogenic potential of estrogens might be attributed to DNA modification caused by derivatives formed during metabolism. $17{\beta}$-estradiol ($E_2$), the main steroidal estrogen present in women, is metabolized via two major pathways: formation of 2-hydroxyestradiol (2-OH $E_2$) and 4-hydroxyestradiol ($4-OH\;E_2$) through the action of cytochrome P450 (CYP) 1A1 and 1B1, respectively. Previous reports suggested that $2-OH\;E_2$ has putative protective effects, while $4-OH\;E_2$ is genotoxic and has potent carcinogenic activity. Thus, the ratio of $2-OH\;E_2/4-OH\;E_2$ is a critical determinant of the toxicity of $E_2$ in mammary cells. In the present study, we investigated the effects of berberine on the expression profile of the estrogen metabolizing enzymes CYP1A1 and CYP1B1 in breast cancer MCF-7 cells. Berberine treatment produced significant induction of both forms at the level of mRNA expression, but with increased doses produced 16~ to 52~fold greater induction of CYP1A1 mRNA over CYP1B1 mRNA. Furthermore, berberine dramatically increased CYP1A1 protein levels but did not influence CYP1B1 protein levels in MCF-7 cells. In conclusion, we present the first report to show that berberine may provide protection against breast cancer by altering the ratio of CYP1A1/CYP1B1, could redirect $E_2$ metabolism in a more protective pathway in breast cancer MCF-7 cells.

      • Exogenous p53 Upregulated Modulator of Apoptosis (PUMA) Decreases Growth of Lung Cancer A549 Cells

        Liu, Chun-Ju,Zhang, Xia-Li,Luo, Da-Ya,Zhu, Wei-Feng,Wan, Hui-Fang,Yang, Jun-Ping,Yang, Xiao-Jun,Wan, Fu-Sheng Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

        Purpose: To investigate the influence of exogenous p53 upregulated modulator of apoptosis (PUMA) expression on cell proliferation and apoptosis in human non-small cell lung cancer A549 cells and transplanted tumor cell growth in nude mice. Materials and Methods: A549 cells were divided into the following groups: control, non-carrier (NC), PUMA (transfected with pCEP4-(HA) 2-PUMA plasmid), DDP ($10{\mu}g/mL$ cisplatin treatment) and PUMA+DDP (transfected with pCEP4-(HA)2-PUMA plasmid and $10{\mu}g/mL$ cisplatin treatment). The MTT method was used to detect the cell survival rate. Cell apoptosis rates were measured by flow cytometry, and PUMA, Bax and Bcl-2 protein expression levels were measured by Western blotting. Results: Compared to the control group, the PUMA, DDP and PUMA+DDP groups all had significantly decreased A549 cell proliferation (p<0.01), with the largest reduction in the PUMA+DDP group. Conversely, the apoptosis rates of the three groups were significantly increased (P<0.01), and the PUMA and DDP treatments were synergistic. Moreover, Bax protein levels significantly increased (p<0.01), while Bcl-2 protein levels significantly decreased (p<0.01). Finally, both the volume and the weights of transplanted tumors were significantly reduced (p<0.01), and the inhibition ratio of the PUMA+DDP group was significantly higher than in the single DDP or PUMA groups. Conclusions: Exogenous PUMA effectively inhibited lung cancer A549 cell proliferation and transplanted tumor growth by increasing Bax protein levels and reducing Bcl-2 protein levels.

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