1.Effect of adenovirus-mediated recombinant Buthus martensii Karsch chloride toxin artifact on human glioma U251 cells
Liangchong CHEN ; Tao HU ; Langlang ZHOU ; Rui HUANG ; Baolai LIU ; Huimin GUO ; Shengli CHEN
Cancer Research and Clinic 2021;33(4):264-269
Objective:To investigate the inhibitory effect of adenovirus-mediated recombinant Buthus martensii Karsch chloride toxin artifact (Ad-rBmK CTa) on human glioma U251 cells and its related mechanisms.Methods:Groups of 3 titer gradients of 3.5×10 9, 7.0×10 9 and 3.5×10 10 pfu/ml Ad-rBmK CTa were set up and applied to U251 cells for 24, 48 and 72 h, and a blank control group (no cells and Ad-rBmK CTa were added) and a negative control group (only U251 cells were added) were set up at the same time. The virus infection status was observed by laser confocal fluorescence microscopy. The cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle and apoptosis in each group were detected by flow cytometry. The expressions of apoptosis-related proteins bax, bcl-2 and caspase-3 were detected by Western blot. Results:The infection rate of Ad-rBmK CTa was over 90% after acting on U251 cells for 24 h. As the titer of Ad-rBmK CTa increased, the proliferation inhibition rate of U251 cells treated for the same hours gradually increased (all P <0.01); with the extension of time, the proliferation inhibition rate of U251 cells treated with the same titer of Ad-rBmK CTa also gradually increased (all P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, the proportion of cells in G 0/G 1 phase was (40.7±0.8)%, and cells in S phase and G 2 phase accounted for (35.7±0.6)% and (23.6±1.4)%, and the difference was statistically significant ( F = 225.119, P < 0.01). When 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 24, 48 and 72 h, the apoptosis rates were (7.4±1.4)%, (19.2±1.7)% and (22.3±1.7)% ( F = 49.470, P < 0.01). After 7.0×10 9 pfu/ml Ad-rBmK CTa acted on U251 cells for 48 h, compared with the negative control group, the expressions of bax and caspase-3 proteins increased, and the expressions of bcl-2 decreased. Conclusions:Ad-rBmK CTa may act on the DNA damage-induced G 1/S detection site to arrest the cell cycle in G 0/G 1 phase, thus inhibiting the proliferation of U251 cells in vitro. However, its induction of apoptosis in U251 cells is not obvious. The mechanism may be related to the direct or indirect inhibition of chloride ion channels.
2.Fingerprint analysis of gamboge by HPLC.
Wenjie HOU ; Baolai CHEN ; Wei XIAO
China Journal of Chinese Materia Medica 2011;36(6):775-779
OBJECTIVETo establish the chromatographic fingerprint for the quality control of gamboge.
METHODAnalysis on a Luna C8 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phases containing acetonitrile and 0.1% glacial acetic acid in water in gradient mode. The flow rate was 1.0 mL x min(-1) and the detection wavelength was at 362 nm. The temperature of column was 25 degrees C. And data of 11 batches of gamboge samples from different sources were analysed by "similarity evaluation for chromatographic Fingerprint of Traditional Chinese Medicine" software.
RESULTThirteen common peaks were selected in chromatograms, and all the common peeks were separated effectively.
CONCLUSIONThe precision, repeatability, and stability of this method were satisfying. The method developed can be used to identify and evaluate the quality of gamboge.
Chromatography, High Pressure Liquid ; methods ; Garcinia ; chemistry ; Quality Control
3.Fingerprint and Multi-components Determination of Saponins in Sanjie-zhentong Capsule
Jianping QIN ; Jianxiong WU ; Jiachun LI ; Baolai CHEN ; Hongmei WEN ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(9):1980-1984
This study was aimed to establish a HPLC fingerprint of saponins in Sanjie-zhentong Capsule in order to make a quantitative analysis of the quality of Sanjie-zhentong Capsule. The Waters Symmetry ShieldTM RP18 (4.6 mmí 250 mm, 5 μm) column was used with a mobile phase of acetonitrile-water gradient elution. The flow rate was 1.2 mL/min. The column temperature was 30℃. The detection wavelength was 203 nm. The results showed that the fingerprint chromatography included 9 mutual peaks. The similarity among batches was more than 0.95. Compared with reference substance, five characteristic components were recognized. The five components are notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd. It was concluded that this method was rapid, simple and accurate and can be used as one of the effective methods for the quality control of Sanjie-zhen-tong Capsule.
4.Determination of Ambrisentan Reference Substance by 1 H-Nuclear Magnetic Resonance Spectroscopy
Yan ZHONG ; Baolai CHEN ; Jiachun LI ; Wenzhe HUANG ; Zhenzhong WANG ; Wei XIAO
China Pharmacist 2015;(5):750-752
Objective:To develop a method for the quantitative determination of ambrisentan. Methods: 1 H NMR spectra were obtalned with a Bruker AscendTM 400 superconducting NMR spectrometer. For each sample, DMSO-D6 was used as the solvent, the pulse width was 10. 0 μs, the delay time was 5 s and the scanning time was 16. Results: The proton peaks of ambrisentan at δ6. 16 ppm and maleic acid atδ6. 28 ppm were used as the quantitative peaks. The linear regression equation of peak area and quality ratio was Y=0. 140 7X+0. 034 8 with the correlation coefficient of 0. 999 4. RSD was 0. 2%(n=6)in the repeated experiments. The absolute content of ambrisentan reference substance was 99. 9%. Conclusion: The results showed that 1 H NMR can be used in the quantitative determination of ambrisentan without reference substance. The method is reliable, rapid, accurate and simple.
5.Identification of Main Related Substances in Baicalein
Qiao ZHANG ; Baolai CHEN ; Xue XIE ; Yiwu ZHAO ; Juan SHEN ; Zhenzhong WANG ; Yang LYU ; Guanhua DU ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1569-1573
This study was aimed to identify the main related substance in baicalein in order to provide basic study data for the safety, efficacy and quality control of the medicine. The analysis was carried out on activated carbon and silica gel column. The compound structure was identified by spectral analysis, which included UV, IR, NMR and MS. The results showed that the related substance was identified as oroxylin A. It was concluded that the determina-tion provided scientific data for studying the impurities of baicalein and improving its quality.
6.A comparative study of targeting drug magnetic nanoparticles and other chemotherapy drugs on human cholangiocarcinoma xenografts
Tao TANG ; Jianwei ZHENG ; Hong LI ; Baolai XIAO ; Bo CHEN ; Gao LI ; Shuai XU ; Shengquan ZHOU ; Xing AI ; Xi LI
Chinese Journal of General Surgery 1993;0(02):-
Objective To study the effect of magnetic nanoparticles on human cholangiocarcinoma xenograft in nude mice, and compared with otherchemotherapy drugs Methods We established human cholangiocarcinoma xenograft in nude mice with QBC939 cell line.The nude mice were devided into 4 groups randomly.Saline,5-FU, Gemcitabine and magnetic nanoparticles were given to nude mice through tail vein on 20d after implanting QBC939 cell line. Calculations were done at different time after treatment in order to compare tumor volume,inhibition ratio of tumor and tumor growth curve of each group. The nude mice were killed on 35d after treatment to harvest tissue for electron microscopic examination to observe ultra-structural changes. Results The tumor volume of control, 5-FU, magnetic nanoparticles and Gemcitabine groups was (2256.1?267.1) mm3, (2096.5?237.9)mm3,(1392.2?189)mm3, and (1534.9?115 )mm3 respectively.The last two groups have significant difference compared to the first two groups(P
7.The expression and significance of iNOS, PDGF-B and LPS in rat models with Budd-Chiari syndrome
Nan ZHU ; Tiantian ZHANG ; Weifu LÜ ; Delei CHENG ; Guangya ZHOU ; Weishi CHEN ; Baolai LIU ; Jiangtao REN ; Dong LU ; Chunze ZHOU
Journal of Interventional Radiology 2019;28(3):262-267
Objective To investigate the expression and significance of inducible nitric oxide synthase (iNOS), platelet-derived growth factor (PDGF)-B and lipopolysaccharide (LPS) in rat models with Budd-Chiari syndrome (BCS) . Methods BCS model was established by partial ligation of inferior vena cava in the posterior segment of the liver. The experimental rats were divided into control group (n=20), model group (n=20) and sham group (n=20) . Liver tissues were collected for immunohistochemistry, HE and Masson staining, and the expression levels of iNOS, PDGF-B and LPS were determined. Results The LPS value in model group was higher than that in both control group and sham group (P=0.001) . The mRNA and protein expressions of iNOS and PDGF-B in model group were higher than those in both control group and sham group (P=0.001) . Statistically significant differences in mRNA and protein expressions of iNOS and PDGF-B existed between each other among the subgroups (P=0.001) . In model group iNOS was positively correlated with PDGF-B and LPS; liver fibrosis was positively correlated with LPS and negatively correlated with PDGFB. Conclusion The damage and repair of BCS is a complicated process. The iNOS, PDGF-B and LPS may play different roles in different stages of BCS. How to regulate their balance in liver fibrosis may be a direction that deserves further study.