To explore the effect of NF κB on bcl x gene transcription in extended drug resistance leukemia cell line HL 60/E6, drug resistant subline HL 60/E6 was derived by intermittently exposing HL 60 cells to 6 ng/ml ...To explore the effect of NF κB on bcl x gene transcription in extended drug resistance leukemia cell line HL 60/E6, drug resistant subline HL 60/E6 was derived by intermittently exposing HL 60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF κB RelA in HL 60/E6 cells. FCM analysis and RT PCR were used to detect the efficiency of liposome mediated ODN transfection and the change of bcl x L mRNA levels after 5 μmol/L phosphorothioate (PS) derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL 60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL 60/E6 cells,but in the cytoplasm of HL 60 cells, the efficiency of liposome mediated ODN transfection was significantly higher than that of null ODN ( P <0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL 60/E6 cells to 5 μmol/L AS PS ODN directed to RelA led to a maximal 40 % decline of bcl x L mRNA levels within 8 h. The inhibition rate of bcl x L mRNA was (15±1.79) %, (28±2.34) %, (40±3.47) %, (20±1.54) % in 4 h, 6 h, 8 h, 15 h, respectively, but it was less than 15 % in control group. It was concluded that NF κB was involved in regulating bcl x transcription. It was suggested that NF κB was an important factor for drug resistance in leukemia cells.展开更多
Objective: To explore the effects and mechanism of glycogen synthase kinase 3β (GSK-313) inhibitor (2'Z,3'E)-6-bromo-indirubin-3'-oxime (BIO) on drug resistance in colon cancer cells. Methods: The colon c...Objective: To explore the effects and mechanism of glycogen synthase kinase 3β (GSK-313) inhibitor (2'Z,3'E)-6-bromo-indirubin-3'-oxime (BIO) on drug resistance in colon cancer cells. Methods: The colon cancer SW480 and SW620 cells were treated with BIO, 5-fluorouracil (5-FU) and BIO/5-FU, separately. Cell cycle distribution, apoptosis level and efflux ability of rhodamine 123 (Rh123) were detected by flow cytometry. The protein expressions of P-glycoprotein (P-gp), multidrug resistance protein 2 (MRP2), thymidylate synthase (TS), β-catenin, E2F-1 and βcl-2 were detected by Western blot. β-catenin and P-gp were stained with double immunofluorescence and observed under a confocal microscope. Results: BIO up-regulated β-catenin, P-gp, MRP2 and TS, enhanced the efflux ability of Rh123, decreased Bcl-2 protein and gave the opposite effect to E2F-1 protein in SW480 and SW620 ceils. Furthermore, BIO significantly inhibited cell apoptosis, increased S and G2/M phase cells, and reduced the cell apoptosis induced by 5-FU in SW480 cells, whereas the effects were slight or not obvious in SW620 cells. Conclusion: GSK-3β was involved in drug resistance regulation, and activation of β-catenin and inhibition of E2F-1 may be the most responsible for the enhancement of 5-FU chemotherapy resistance induced by GSK-β inhibitor β10 in colon cancer.展开更多
[Objective]The paper was to understand the resistance of commonly used clinical antibiotics to avian Escherichia coli and the inhibition of lactobacillus against pathogenic E.coli.[Method]The strain was isolated and i...[Objective]The paper was to understand the resistance of commonly used clinical antibiotics to avian Escherichia coli and the inhibition of lactobacillus against pathogenic E.coli.[Method]The strain was isolated and identified by SS medium culture,microscope examination,PCR amplification and biochemical test,and the sensitivity of avian E.coli to eight antibiotics was determined by disk diffusion method.Twelve strains of avian E.coli and one strain of lactobacillus were divided into five group using in vitro liquid culture method,namely simultaneous mixed culture group of lactobacillus and E.coli,first colonization group of E.coli,first colonization group of lactobacillus,E.coli control group and lactobacillus control group,for competitive inhibition test between E.coli and lactobacillus.[Result]Totally 34 strains with similar characteristics to E.coli strains were preliminarily screened;the target band amplified by PCR was 293 bp,which was consistent with the expected size.The isolated strain could decompose maltose,sucrose,lactose,glucose,arabinose,mannitol and sorbitol;the nitrate reduction test was positive,while citrate and V-P test was negative,consistent with the biochemical characteristics of E.coli.The resistance rate of the isolates to cefalexin and tylosin was over 65%,and that to enrofloxacin and doxycycline was up to 50%;the isolates were more sensitive to apramycin,tilmicosin,neomycin and colistin,and the total resistance rate was below 15%.The inhibitory effects of mixed culture of lactobacillus and E.coli against E.coli was better than first colonization of lactobacillus at 24 h;the inhibitory effects of first colonization of lactobacillus against E.coli was better than that of mixed culture of lactobacillus and E.coli at 36 h;the inhibitory effects of mixed culture of lactobacillus and E.coli against E.coli was the best at 48 h.[Conclusion]The isolated strains of E.coli were generally resistant to eight commonly used antibiotics.Lactobacillus could inhibit the growth of avian E.coli in vitro,and first colonization of lactobacillus received the best effort.展开更多
Heparosan is a natural precursor of heparin biosynthesis in mammals. It is stable in blood circulation but can be degraded in lysosomes, showing good biocompatibility and long circulation features. So heparosan can be...Heparosan is a natural precursor of heparin biosynthesis in mammals. It is stable in blood circulation but can be degraded in lysosomes, showing good biocompatibility and long circulation features. So heparosan can be designed as anticancer drug carriers to increase tumor selectivity and improve the therapeutic effect. A novel redox-sensitive heparosancystamine-vitamin E succinate(KSV) micelle system was constructed for intracellular delivery of doxorubicin(DOX). Simultaneously, the redox-insensitive heparosan-adipic acid dihydrazide-vitamin E succinate copolymer(KV) was synthesized as control. DOX-loaded micelles(DOX/KSV) with an average particle size of 90–120 nm had good serum stability and redox-triggered depolymerization. In vitro drug release test showed that DOX/KSV micelles presented obvious redox-triggered release behavior compared with DOX/KV. Cytotoxicity and cell uptake were investigated using MGC80-3 tumor cells and COS7 fibroblast-like cells. The cell survival rate of blank micelles was more than 90%, and the cytotoxicity of DOX/KSV in MGC80-3 cells was higher than in COS7 cells, indicating that the carrier has better biocompatibility and less toxicity side effect. The cytotoxicity of DOX/KSV against MGC80-3 cells was significantly greater than that of free DOX and DOX/KV. Furthermore, compared with DOX/KV in MGC80-3 cells, DOX/KSV micelles uptook more anticancer drugs and then released DOX faster into the cell nucleus. The micelles were endocytosed by multiple pathways, but clathrin-mediated endocytosis was the main pathway. Therefore, heparosan polysaccharide could be a potential option as anticancer carrier for enhancing efficacy and mitigating toxicity.展开更多
Background and Objective: miRNA-200c can not only inhibit the aggressiveness of cancer cells but also increase the sensitivity of cells to antitumor drugs. However, some mechanisms are still unclear. Recent researches...Background and Objective: miRNA-200c can not only inhibit the aggressiveness of cancer cells but also increase the sensitivity of cells to antitumor drugs. However, some mechanisms are still unclear. Recent researches revealed that E-cadherin is more than an inhibitor of metastasis, and it also plays important roles in reversing drug resistance. We had previously found that miRNA-200c could not only induce the expression of E-cadherin but also increase the sensitivity of gastric cancer SGC7901/DDP cells to cisplatin (DDP). This study aimed to explore the effects of miRNA-200c on biological characteristics of SGC7901/DDP cells and the roles of E-cadherin in the regulatory pathway of miRNA-200c. Methods: SGC7901/DDP cells and its parental cell line SGC7901 cells were transfected with miRNA-200c precursor (Pre-200c) and E-cadherin siRNA, respectively. Real-time RT-PCR was used to detect miRNA-200c expression after transfection with Pre-200c in SGC7901/DDP cell line. Drug sensitivities to DDP, 5-fluorouracil (5-FU), paclitaxel, and adriamycin (ADR) after transfection were tested using MTT assay. The proliferation of SGC7901/DDP cells was also detected after transfection. The protein changes of E-cadherin, Bax, and Bcl-2 after transfection were detected by Western blot. Results: The miRNA-200c expression in SGC7901/DDP cells after transfection of Pre-200c was 7.128 ± 0.159 times of that in negative control (P < 0.05). The IC50 of DDP, 5-FU, paclitaxel, and ADR in Pre-200c-transfected group were significantly lower than that in negative control group (P < 0.05). Compared to the control group, cell proliferation was significantly decreased (P < 0.05). The relative protein expressions of E-cadherin and Bax in Pre-200c-transfected group were significantly higher than those in negative control group (P < 0.05), whereas Bcl-2 was significantly lower than that in control (P < 0.05). Additionally, E-cadherin protein expression was significantly inhibited after transfected with E-cadherin siRNA in SGC7901 cells. The Bax protein expression was significantly down-regulated by E-cadherin siRNA (P < 0.05), whereas the Bcl-2 expression was significantly up-regulated (P < 0.05). Conclusion: miRNA-200c can indirectly regulate apoptosis through E-cadherin in SGC7901/DDP cells, which may be a possible mechanism of miRNA-200c in reversing drug resistance and inhibiting proliferation.展开更多
目的基于Web of Science数据库,利用文献计量学和可视化分析探索多黏菌素E治疗药物监测(TDM)相关研究的现状、热点和发展趋势。方法收集Web of Science核心数据库2012至2022年收录的多黏菌素E TDM相关研究的文献,应用机器学习和可视化工...目的基于Web of Science数据库,利用文献计量学和可视化分析探索多黏菌素E治疗药物监测(TDM)相关研究的现状、热点和发展趋势。方法收集Web of Science核心数据库2012至2022年收录的多黏菌素E TDM相关研究的文献,应用机器学习和可视化工具,包括VOSviewer软件、书目共现分析系统和图形聚类工具包进行文献计量学分析。结果本研究共纳入713篇文献,2012至2022年文献的发表量有明显上升。在该领域贡献最多的国家和研究者分别是美国和LI J团队。多黏菌素E是出现最多的关键词,并与肾毒性和药物代谢动力学关联度最高。研究领域包括了多黏菌素E剂量与肾毒性的关系、联合给药的抗菌活性及耐药性、治疗监测与药代动力学。群体药代动力学和相关的模型是最新的研究热点。结论本研究文献计量分析能更好地了解多黏菌素E TDM相关研究的现状,并对未来研究进行了展望,未来应加快推进多黏菌素E TDM室间质控和评价研究,完善质控方法。此外应对硫酸黏菌素群体药代动力学及剂量优化进行更多的探索研究。展开更多
Poly(e-caprolactone) (PCL) with weight-average molar mass over 10000 g/mol was synthesized by microwave-assisted ring-opening polymerization of e-caprolactone (e-CL) with maleic acid (MA) as initiator (2.45 GHz, 360 W...Poly(e-caprolactone) (PCL) with weight-average molar mass over 10000 g/mol was synthesized by microwave-assisted ring-opening polymerization of e-caprolactone (e-CL) with maleic acid (MA) as initiator (2.45 GHz, 360 W, 85 min). Ibuprofen-PCL controlled release system was prepared directly by the ROP of e-CL in its mixture with ibuprofen. The release of ibuprofen from the system was sustained and steady.展开更多
文摘To explore the effect of NF κB on bcl x gene transcription in extended drug resistance leukemia cell line HL 60/E6, drug resistant subline HL 60/E6 was derived by intermittently exposing HL 60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF κB RelA in HL 60/E6 cells. FCM analysis and RT PCR were used to detect the efficiency of liposome mediated ODN transfection and the change of bcl x L mRNA levels after 5 μmol/L phosphorothioate (PS) derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL 60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL 60/E6 cells,but in the cytoplasm of HL 60 cells, the efficiency of liposome mediated ODN transfection was significantly higher than that of null ODN ( P <0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL 60/E6 cells to 5 μmol/L AS PS ODN directed to RelA led to a maximal 40 % decline of bcl x L mRNA levels within 8 h. The inhibition rate of bcl x L mRNA was (15±1.79) %, (28±2.34) %, (40±3.47) %, (20±1.54) % in 4 h, 6 h, 8 h, 15 h, respectively, but it was less than 15 % in control group. It was concluded that NF κB was involved in regulating bcl x transcription. It was suggested that NF κB was an important factor for drug resistance in leukemia cells.
基金supported by the Sci-Tech Project Foundation of Guangdong Province(No.2009B080800023)
文摘Objective: To explore the effects and mechanism of glycogen synthase kinase 3β (GSK-313) inhibitor (2'Z,3'E)-6-bromo-indirubin-3'-oxime (BIO) on drug resistance in colon cancer cells. Methods: The colon cancer SW480 and SW620 cells were treated with BIO, 5-fluorouracil (5-FU) and BIO/5-FU, separately. Cell cycle distribution, apoptosis level and efflux ability of rhodamine 123 (Rh123) were detected by flow cytometry. The protein expressions of P-glycoprotein (P-gp), multidrug resistance protein 2 (MRP2), thymidylate synthase (TS), β-catenin, E2F-1 and βcl-2 were detected by Western blot. β-catenin and P-gp were stained with double immunofluorescence and observed under a confocal microscope. Results: BIO up-regulated β-catenin, P-gp, MRP2 and TS, enhanced the efflux ability of Rh123, decreased Bcl-2 protein and gave the opposite effect to E2F-1 protein in SW480 and SW620 ceils. Furthermore, BIO significantly inhibited cell apoptosis, increased S and G2/M phase cells, and reduced the cell apoptosis induced by 5-FU in SW480 cells, whereas the effects were slight or not obvious in SW620 cells. Conclusion: GSK-3β was involved in drug resistance regulation, and activation of β-catenin and inhibition of E2F-1 may be the most responsible for the enhancement of 5-FU chemotherapy resistance induced by GSK-β inhibitor β10 in colon cancer.
基金Major Science and Technology Innovation Project of Liaoning Province"Feed Quality and Safety Control and Low Protein Feed Project"(2019JH1/10200002).
文摘[Objective]The paper was to understand the resistance of commonly used clinical antibiotics to avian Escherichia coli and the inhibition of lactobacillus against pathogenic E.coli.[Method]The strain was isolated and identified by SS medium culture,microscope examination,PCR amplification and biochemical test,and the sensitivity of avian E.coli to eight antibiotics was determined by disk diffusion method.Twelve strains of avian E.coli and one strain of lactobacillus were divided into five group using in vitro liquid culture method,namely simultaneous mixed culture group of lactobacillus and E.coli,first colonization group of E.coli,first colonization group of lactobacillus,E.coli control group and lactobacillus control group,for competitive inhibition test between E.coli and lactobacillus.[Result]Totally 34 strains with similar characteristics to E.coli strains were preliminarily screened;the target band amplified by PCR was 293 bp,which was consistent with the expected size.The isolated strain could decompose maltose,sucrose,lactose,glucose,arabinose,mannitol and sorbitol;the nitrate reduction test was positive,while citrate and V-P test was negative,consistent with the biochemical characteristics of E.coli.The resistance rate of the isolates to cefalexin and tylosin was over 65%,and that to enrofloxacin and doxycycline was up to 50%;the isolates were more sensitive to apramycin,tilmicosin,neomycin and colistin,and the total resistance rate was below 15%.The inhibitory effects of mixed culture of lactobacillus and E.coli against E.coli was better than first colonization of lactobacillus at 24 h;the inhibitory effects of first colonization of lactobacillus against E.coli was better than that of mixed culture of lactobacillus and E.coli at 36 h;the inhibitory effects of mixed culture of lactobacillus and E.coli against E.coli was the best at 48 h.[Conclusion]The isolated strains of E.coli were generally resistant to eight commonly used antibiotics.Lactobacillus could inhibit the growth of avian E.coli in vitro,and first colonization of lactobacillus received the best effort.
基金We are grateful for the financial support from National Natural Science Foundation of China(81503007 and 21574059)Research Project of Wuxi Health and Family Planning Commission(Q201843)+1 种基金Natural Science Foundation of Jiangsu Province(BK20170202)the Top-notch Academic Programs Project of Jiangsu Higher Education Institutions(PPZY2015B146).
文摘Heparosan is a natural precursor of heparin biosynthesis in mammals. It is stable in blood circulation but can be degraded in lysosomes, showing good biocompatibility and long circulation features. So heparosan can be designed as anticancer drug carriers to increase tumor selectivity and improve the therapeutic effect. A novel redox-sensitive heparosancystamine-vitamin E succinate(KSV) micelle system was constructed for intracellular delivery of doxorubicin(DOX). Simultaneously, the redox-insensitive heparosan-adipic acid dihydrazide-vitamin E succinate copolymer(KV) was synthesized as control. DOX-loaded micelles(DOX/KSV) with an average particle size of 90–120 nm had good serum stability and redox-triggered depolymerization. In vitro drug release test showed that DOX/KSV micelles presented obvious redox-triggered release behavior compared with DOX/KV. Cytotoxicity and cell uptake were investigated using MGC80-3 tumor cells and COS7 fibroblast-like cells. The cell survival rate of blank micelles was more than 90%, and the cytotoxicity of DOX/KSV in MGC80-3 cells was higher than in COS7 cells, indicating that the carrier has better biocompatibility and less toxicity side effect. The cytotoxicity of DOX/KSV against MGC80-3 cells was significantly greater than that of free DOX and DOX/KV. Furthermore, compared with DOX/KV in MGC80-3 cells, DOX/KSV micelles uptook more anticancer drugs and then released DOX faster into the cell nucleus. The micelles were endocytosed by multiple pathways, but clathrin-mediated endocytosis was the main pathway. Therefore, heparosan polysaccharide could be a potential option as anticancer carrier for enhancing efficacy and mitigating toxicity.
文摘Background and Objective: miRNA-200c can not only inhibit the aggressiveness of cancer cells but also increase the sensitivity of cells to antitumor drugs. However, some mechanisms are still unclear. Recent researches revealed that E-cadherin is more than an inhibitor of metastasis, and it also plays important roles in reversing drug resistance. We had previously found that miRNA-200c could not only induce the expression of E-cadherin but also increase the sensitivity of gastric cancer SGC7901/DDP cells to cisplatin (DDP). This study aimed to explore the effects of miRNA-200c on biological characteristics of SGC7901/DDP cells and the roles of E-cadherin in the regulatory pathway of miRNA-200c. Methods: SGC7901/DDP cells and its parental cell line SGC7901 cells were transfected with miRNA-200c precursor (Pre-200c) and E-cadherin siRNA, respectively. Real-time RT-PCR was used to detect miRNA-200c expression after transfection with Pre-200c in SGC7901/DDP cell line. Drug sensitivities to DDP, 5-fluorouracil (5-FU), paclitaxel, and adriamycin (ADR) after transfection were tested using MTT assay. The proliferation of SGC7901/DDP cells was also detected after transfection. The protein changes of E-cadherin, Bax, and Bcl-2 after transfection were detected by Western blot. Results: The miRNA-200c expression in SGC7901/DDP cells after transfection of Pre-200c was 7.128 ± 0.159 times of that in negative control (P < 0.05). The IC50 of DDP, 5-FU, paclitaxel, and ADR in Pre-200c-transfected group were significantly lower than that in negative control group (P < 0.05). Compared to the control group, cell proliferation was significantly decreased (P < 0.05). The relative protein expressions of E-cadherin and Bax in Pre-200c-transfected group were significantly higher than those in negative control group (P < 0.05), whereas Bcl-2 was significantly lower than that in control (P < 0.05). Additionally, E-cadherin protein expression was significantly inhibited after transfected with E-cadherin siRNA in SGC7901 cells. The Bax protein expression was significantly down-regulated by E-cadherin siRNA (P < 0.05), whereas the Bcl-2 expression was significantly up-regulated (P < 0.05). Conclusion: miRNA-200c can indirectly regulate apoptosis through E-cadherin in SGC7901/DDP cells, which may be a possible mechanism of miRNA-200c in reversing drug resistance and inhibiting proliferation.
文摘目的基于Web of Science数据库,利用文献计量学和可视化分析探索多黏菌素E治疗药物监测(TDM)相关研究的现状、热点和发展趋势。方法收集Web of Science核心数据库2012至2022年收录的多黏菌素E TDM相关研究的文献,应用机器学习和可视化工具,包括VOSviewer软件、书目共现分析系统和图形聚类工具包进行文献计量学分析。结果本研究共纳入713篇文献,2012至2022年文献的发表量有明显上升。在该领域贡献最多的国家和研究者分别是美国和LI J团队。多黏菌素E是出现最多的关键词,并与肾毒性和药物代谢动力学关联度最高。研究领域包括了多黏菌素E剂量与肾毒性的关系、联合给药的抗菌活性及耐药性、治疗监测与药代动力学。群体药代动力学和相关的模型是最新的研究热点。结论本研究文献计量分析能更好地了解多黏菌素E TDM相关研究的现状,并对未来研究进行了展望,未来应加快推进多黏菌素E TDM室间质控和评价研究,完善质控方法。此外应对硫酸黏菌素群体药代动力学及剂量优化进行更多的探索研究。
基金This work was financially supported by the Research Foundation of MOE and National 973 Project of China (G1999064703).
文摘Poly(e-caprolactone) (PCL) with weight-average molar mass over 10000 g/mol was synthesized by microwave-assisted ring-opening polymerization of e-caprolactone (e-CL) with maleic acid (MA) as initiator (2.45 GHz, 360 W, 85 min). Ibuprofen-PCL controlled release system was prepared directly by the ROP of e-CL in its mixture with ibuprofen. The release of ibuprofen from the system was sustained and steady.