1.In vitro study on mesothelin antibody-tagged nanoprobe targeting human pancreatic cell line BxPC3
Mingzhi LU ; Wenjun LE ; Shaobin CUI ; Bingmei SUN ; Bingdi CHEN ; Chengwei SHAO
Chinese Journal of Pancreatology 2014;14(6):389-391
Objective To observe the targeted function of a mesothelin antibody modified nanoprobe in human pancreatic cancer BxPC3 cell.Methods The Fe3O4@SiO2 nanoprobe was prepared by St(o)ber method,and then quantum dots (CdTe) and mesothelin antibody was crosslinked to obtain the properties of targeting and fluorescent.Fluorescent nano Fe3O4@SiO2 probes and BxPC3 cells were incubated in vitro for 30 min.Its targeting performance was tested by the CCD imaging system and magnetic separation technology.HepG-2 and K562 cells with low expression of mesothelin were selected as reference cells.Results This preparation method of nanoprobe could produce a uniform and narrow distribution particle with particle size mainly ranging from 120 to 140 nm.The cell adsorption experiments showed that the adsorption efficiency of BxPC3,HepG-2 and K562 by nanoprobe without crosslinking antibody were less than 20%,as a non-specific adsorption; and the adsorption efficiency of BxPC3,HepG-2 and K562 by crosslinking mesothelin antibody nanoprobe were (53.9 ± 1.8) %,(8.0 ± 2.1) % and (8.9 ± 2.3) % respectively,and the adsorption capacity with BxPC3 was significantly increased.Conclusions The nanoprobe modified by mesothelin antibody can effectively recognize BxPC3 cells which highly expressing mesothelin.
3.Establishment of UPLC fingerprint and study on anti-respiratory syncytial virus spectrum-effective relationship of ethanol extract from Callicarpa nudiflora
Bingmei SHAO ; Xiaofeng LI ; Beibei YAN ; Qifeng LIU ; Huanyun QIN ; Xiangjun ZHAO ; Lin HOU
China Pharmacy 2023;34(20):2465-2470
OBJECTIVE To establish the fingerprint of the ethanol extract from Callicarpa nudiflora, analyze its anti- respiratory syncytial virus (RSV) activity in vitro, and study the relationship between spectrum and effect. METHODS Using 10%, 30%, 50%, 70% and 90% ethanol as solvent, 20 batches of ethanol extracts from 4 batches of C. nudiflora were prepared. The fingerprints for 20 batches of ethanol extracts from C. nudiflora were mapped by ultra-high-performance liquid chromatography (UPLC), and the similarity evaluation was conducted by using the Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints (2012 edition). The cytopathic effect method and MTT method were used to investigate the in vitro inhibitory activity of the ethanol extracts from C. nudiflora on RSV. Pearson correlation analysis, grey correlation degree and orthogonal partial least squares (OPLS) analysis were used to study the spectrum-effect relationship. RESULTS There were 25 common peaks in 20 batches of ethanol extracts from C. nudiflora, and the similarities ranged from 0.912 to 0.998, and the RSDs of common peak areas were 33.54%-162.28%. The average values of IC50 for RSV of 20 batches of ethanol extracts from C. nudiflora were 9.55-272.23 μg/mL. The results of Pearson correlation analysis, grey correlation analysis and OPLS analysis showed that the Pearson correlation coefficients (P<0.05) of the common peaks 8, 10, 12, 16, 18-19, 22-24 with pharmacodynamic indicators and regression coefficients were all negative, the correlation coefficients were all greater than 0.6, and the values of variable importance in projection were all greater than 1. CONCLUSIONS Twenty batches of ethanol extracts from C. nudiflora have similar components but significant differences in content, and exhibit different degrees of anti-RSV activity in vitro. The corresponding components of common peaks 8, 10, 12, 16, 18-19, 22-24 may be the characteristic components of anti-RSV of C. nudiflora.
4.Study on the differential markers of Morus alba leaves before and after baked with honey based on fingerprint and chemometrics
Beibei YAN ; Xiaoping ZHANG ; Huanyun QIN ; Xiangjun ZHAO ; Bingmei SHAO ; Jingzhen TIAN ; Dalong MA
China Pharmacy 2024;35(10):1198-1202
OBJECTIVE To compare the changes of chemical components of Morus alba leaves, screen differential markers, and determine their contents, so as to provide reference for quality control of M. alba leaves before and after baked with honey. METHODS The fingerprints of M. alba leaves before and after baked with honey were established by high-performance liquid chromatography (HPLC). The common peaks of the fingerprints were identified and the similarity was evaluated. The differential markers of M. alba leaves before and after baked with honey were screened by principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) using common peak are of raw material and product baked with honey of M. alba leaves as index. The quantitative analysis was carried out. RESULTS Twenty-three and twenty-four common peaks were identified from the HPLC fingerprint spectra of ten batches of raw material and ten batches of product baked with honey of M. alba leaves, respectively. The similarities of HPLC fingerprints for raw material and product baked with honey of M. alba leaves were all greater than 0.97. The results of PCA showed that raw material and product baked with honey of M. alba leaves could be divided into two categories. The results of OPLS-DA showed that the variable importance in projection of peak 2, peak H (5- hydroxymethylfurfural), peak 1, peak 17 (isochlorogenic acid C) and peak 16 were all greater than 1. The average contents of differential marker of isochlorogenic acid C in raw material and product baked with honey of M. alba leaves were 0.093 6 and 0.127 8 mg/g, respectively; there was statistical significance (P<0.05). CONCLUSIONS Five differential markers such as isochlorogenic acid C are obtained. The content of isochlorogenic acid C in M. alba leaves is significantly increased after baked with honey.