ABSTRACT
Objective To investigate preventive and therapeutic effects ofAngong Niuhuang Pills (ANP) on cerebrovascular diseases in Zebrafish models.Methods Zebrafish cerebral hemorrhage was induced by treatment with simvastatin,thrombosis by arachidonic acid,blood vessel loss by simvastatin,and Alzheimer's disease (AD) by A1C13.Zebrafish models were treated with ANP by direct drug soaking at various concentrations.Results ANP had a significantly preventive effect on Zebrafish cerebral hemorrhage with an efficacy of 11％,41％,and 48％ at concentrations of 27.8,83.3,and 250 μg/mL,respectively,and the incidence of cerebral hemorrhage in 83.3 and 250 g/mL groups decreased significantly compared with model group (P ＜ 0.001).Compared with model group,the red cell staining intensity of Zebrafish in 333 and 1 000 g/mL groups increased significantly (P ＜ 0.001),and the thrombus prevention rates of 111,333,and 1000 g/mL groups were 5％,33％,and 64％ respectively.Compared with model group,the intestinal vascular area of Zebrafish in ANP 11.1,33.3,and 100.0 μg/mL groups increased significantly (P ＜ 0.001),and the rate of angiogenesis was 21％,25％ and 26％ respectively.ANP promoted dyskinesia recovery by 48％,88％,and 78％,and improved response efficiency by 15％,82％ and 75％,respectively in AD zebrafish at concentrations of 111,333,and 1000 μg/mL.Conclusions Our results in this study support ANP as a preventive and therapeutic medicine for cerebral hemorrhage,cerebral ischemia,thrombosis and AD.
ABSTRACT
Objective To construct the eukaryotic expression vector of human BRE1B labeled with pCMV-Tag-2B and detect its biological activity in melanoma cells preliminarily.Methods Ocular B16 melanoma cells were randomly divided into the experimental group,in which the cells were transfected with pCMV-Tag-2B-BRE1B and the control group,which was transfected with pCMV plasmid.The CDS coding region of human BRE1B gene was amplified by PCR using human mammary gland cDNA as a template for construction of the recombinant plasmid pCMV-Tag-2B-BRE1B.After transfected with pCMV-Tag-2B-BRE1B and pCMV plasmid in the experimental and control group,respectively,Western blot was applied to detect the expression of BRE1B protein,while cell counting kit-8 (CCK8) and colony assays were used to analyze the effects of recombinant plasmid pCMV-Tag-2B-BRE1B on the growth of B16 melanoma cells.Results The CDS coding sequence of human BRE1B gene was amplified by PCR successfully,which was equal to the expected size.Compared with the control group,the sequence from bacteria PCR was identified as positive,with the length of 4000 bp and 3050 bp by double enzyme digestion respectively.Moreover,the coding sequence of the human BRE1B gene was exactly the same as the inserted DNA sequence.Western blot results showed that the expression of recombinant plasmid pCMV-Tag-2B-BRE1B was successfully expressed in the experimental group,but there was no specific fragments in the control group.And cell counting kit-8 (CCK8) and colony assays showed that pCMV-Tag-2B-BRE1B recombinant plasmid could inhibit the growth of B16 melanoma cells.Conclusion The eukaryotic expression vector of pCMV-Tag-2B-BRE1B labeled with pCMV-Tag-2B is constructed successfully,and it has inhibitory effects on the growth of ocular B16 melanoma cells.
ABSTRACT
Objective To evaluate the sensitivity to 5 clinically commonly used anticancer drugs in vivo using the zebrafish xenotransplantation models of human lung cancer,stomach cancer,and liver cancer cells,respectively. Methods Zebrafish xenotransplantation models of A549 lung cancer cells,SGC-7901 stomach cancer cells and HepG2 liver cancer cells were established. The xenograft models of A549 cells were treated with three different doses of cis-platinum, paclitaxel, vinorelbine, endostar and bevacizumab, respectively. The SGC-7901 model was treated with three concentrations or doses of paclitaxel, irinotecan, hydroxyurea, cis-platinum and 5-fluorouracil, respectively. And the HepG2 model was treated with three concentrations or doses of adriamycin,gemcitabine,hydroxyurea,cis-platinum and 5-fluorouracil. The tumors were analyzed and quantified in vivo by fluorescence microscopy,and the inhibition rates of tumor growth with each drug were calculated and compared with the model control group for statistical significance. Results All of the tested anticancer drugs showed inhibitory effect on tumor cells in the zebrafish xenograft models with statistical significance in a dose-dependent manner. During the drug sensitivity test,the inhibition rate of bevacizumab on A549 lung cancer cells decreased in the order(65%)> cis-platinum(55%)> vinorelbine(40%)> endostar(39%)>paclitaxel(27%). As for the SGC-7901 stomach cancer cells, the tumor growth inhibition rate decreased in the order hydroxyurea(46%)> 5-FU(31%)= irinotecan(31%)> paclitaxel(26%)> cis-platinum(24%). And the therapeutic effect of cis-platinum on the HepG2 liver cancer cells decreased in the order(64%)> hydroxyurea(56%)>gemcitabine(46%)> adriamycin(45%)> 5-FU(38%). Conclusions Zebrafish xenotransplantation models of cancer cells are suitable for in vivo sensitivity test of anticancer drugs.
ABSTRACT
Objective To investigate preventive and therapeutic effects ofAngong Niuhuang Pills (ANP) on cerebrovascular diseases in Zebrafish models.Methods Zebrafish cerebral hemorrhage was induced by treatment with simvastatin,thrombosis by arachidonic acid,blood vessel loss by simvastatin,and Alzheimer's disease (AD) by A1C13.Zebrafish models were treated with ANP by direct drug soaking at various concentrations.Results ANP had a significantly preventive effect on Zebrafish cerebral hemorrhage with an efficacy of 11％,41％,and 48％ at concentrations of 27.8,83.3,and 250 μg/mL,respectively,and the incidence of cerebral hemorrhage in 83.3 and 250 g/mL groups decreased significantly compared with model group (P ＜ 0.001).Compared with model group,the red cell staining intensity of Zebrafish in 333 and 1 000 g/mL groups increased significantly (P ＜ 0.001),and the thrombus prevention rates of 111,333,and 1000 g/mL groups were 5％,33％,and 64％ respectively.Compared with model group,the intestinal vascular area of Zebrafish in ANP 11.1,33.3,and 100.0 μg/mL groups increased significantly (P ＜ 0.001),and the rate of angiogenesis was 21％,25％ and 26％ respectively.ANP promoted dyskinesia recovery by 48％,88％,and 78％,and improved response efficiency by 15％,82％ and 75％,respectively in AD zebrafish at concentrations of 111,333,and 1000 μg/mL.Conclusions Our results in this study support ANP as a preventive and therapeutic medicine for cerebral hemorrhage,cerebral ischemia,thrombosis and AD.