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学者姓名：贺浪冲
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Abstract ：
With the rapid development of therapeutic monoclonal antibody drugs, it is increasingly difficult to meet clinical needs using traditional antibody purification techniques. In this study, epidermal growth factor receptor (EGFR)SNAP-tag was expressed in HEK293 cells. Then we captured the EGFR-SNAP-tag from the cell membrane suspension onto a O-6-benzylguanine-modified silica gel to prepare a new EGFR stationary phase as a bioactive material, which could specifically recognize its antibody in bio-samples. The EGFR stationary phase was systematically characterized via scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, and fourier transform infrared spectroscopy. Then we used EGFR stationary phase to establish a new EGFR cell membrane chromatography (CMC) model. The EGFR/CMC-online-ion exchange chromatography (IEC)/high performance liquid chromatography (HPLC) was established for the efficient purification and specific identification of cetuximab, nituzumab, and panizumab from cell culture medium and human serum. The results show that the EGFR stationary phase prepared by one-step immobilized technique can maintain biological activity and stability like EGFR in cell membrane. The EGFR/CMC-online-IEC/HPLC method has a high specificity, accuracy and sensitivity. Therefore, it will present a valuable method for the purification, identification, and analysis of monoclonal antibody drugs.
Keyword ：
Antibody purification cell Epidermal growth factor receptor membrane chromatography SNAP-tag
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| GB/T 7714 | Fu, Jia , Lv, Yanni , Jia, Qianqian et al. Purification and determination of antibody drugs in bio-samples by EGFR/ cell membrane chromatography method [J]. | JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS , 2022 , 217 . |
| MLA | Fu, Jia et al. "Purification and determination of antibody drugs in bio-samples by EGFR/ cell membrane chromatography method" . | JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 217 (2022) . |
| APA | Fu, Jia , Lv, Yanni , Jia, Qianqian , Wang, Cheng , Wang, Saisai , Liang, Peida et al. Purification and determination of antibody drugs in bio-samples by EGFR/ cell membrane chromatography method . | JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS , 2022 , 217 . |
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Abstract ：
The determination of fat-soluble vitamins and carotenoids in human serum provides reliable information for diagnosing malnutrition and for establishing appropriate intervention programs. Due to the complex composition of the biological samples, the efficient sample preparation is the key to the analysis. We report here a surface active ionic liquid (SAIL)-based dispersive liquid-liquid microextraction (DLLME) method coupled with a high performance liquid chromatography (HPLC) to determine four fat-soluble vitamins and six carotenoids in human serum simultaneously. Liquid crystal structures were formed in the extract phase. And the enrichment factor of the analytes treated by DLLME was 4 to 26 times of the traditional LLE method except lycopene. The limit of determination for these compounds was determined to be between 0.002 and 0.076 mu g/mL. The accuracy was validated by the standard addition method with recoveries ranging from 82.4 to 114.1%. The intra-day and inter-day relative standard deviations were 2.76-12.63% and 4.01-13.54%, respectively. The proposed DLLME coupled with the HPLC method was successfully applied in the determination of fat-soluble micronutrients in human serum. (c) 2022 Elsevier B.V. All rights reserved.
Keyword ：
Carotenoids DLLME Fat-soluble vitamins Human serum Surface-active ionic liquid
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| GB/T 7714 | Kong, Liyun , Wang, Jiaqi , Gao, Qingpeng et al. Simultaneous determination of fat-soluble vitamins and carotenoids in human serum using a nanostructured ionic liquid based microextraction method [J]. | JOURNAL OF CHROMATOGRAPHY A , 2022 , 1666 . |
| MLA | Kong, Liyun et al. "Simultaneous determination of fat-soluble vitamins and carotenoids in human serum using a nanostructured ionic liquid based microextraction method" . | JOURNAL OF CHROMATOGRAPHY A 1666 (2022) . |
| APA | Kong, Liyun , Wang, Jiaqi , Gao, Qingpeng , Li, Xiaoqian , Zhang, Wenbin , Wang, Ping et al. Simultaneous determination of fat-soluble vitamins and carotenoids in human serum using a nanostructured ionic liquid based microextraction method . | JOURNAL OF CHROMATOGRAPHY A , 2022 , 1666 . |
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Abstract ：
Only a few systematic tack force studies have been conducted on poultices although they are widely used in therapeutic fields. In this study, we evaluated the factors influencing the tack force of poultices using the design of experiments (DoE) approach. The effects of detachment speed, pressuring time, pressuring force, coating thickness, carbomer content, and active pharmaceutical integrant (API) content on adhesion performance were evaluated using 46 response surface designs, after which the maximum value of the stress curve (sigma(max)) was recorded. The results revealed that all factors except pressuring time significantly affected the response (at the 5% level), and two-factor interactions were identified using the DoE approach to evaluate carbomer content, pressuring force, coating thickness, and API content.
Keyword ：
Indomethacin poultices multi-factor probe tack force
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| GB/T 7714 | Chen, Hua , Ma, Xun , Xu, Daiyue et al. Investigation of the tack force of poultices using the probe tack test [J]. | JOURNAL OF ADHESION SCIENCE AND TECHNOLOGY , 2022 . |
| MLA | Chen, Hua et al. "Investigation of the tack force of poultices using the probe tack test" . | JOURNAL OF ADHESION SCIENCE AND TECHNOLOGY (2022) . |
| APA | Chen, Hua , Ma, Xun , Xu, Daiyue , Chen, Guang , He, Langchong . Investigation of the tack force of poultices using the probe tack test . | JOURNAL OF ADHESION SCIENCE AND TECHNOLOGY , 2022 . |
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Pseudo-allergic reactions (PARs) are IgE-independent hypersensitivity reactions. Mas-related G protein-coupled receptor-X2 (MrgX2) was proved the key receptor of PAR. The anti-pseudo-allergic compound discovery based on MrgX2 was of great value. Cell membrane chromatography (CMC) based on MrgX2 provides a convenient and effective tool in anti-pseudoallergic compound screening and discovery, and further improvements of this method are still needed. In this work, SNAP-tag was introduced at C-terminal of Mas-related G protein-coupled receptor (MrgX2-SNAP-tag), and an MrgX2-SNAP-tag/ CMC model was then conducted using CMC technique. Comparative experiments showed that the new model not only satisfied the good selectivity and specificity of screening but also exhibited more stable and longer life span than traditional MrgX2/CMC model. By coupling with HPLC-MS, two compounds were screened out from Arnebiae Radix and identified as shikonin and acetylshikonin. Nonlinear chromatography was performed to study the interactions between two screened compounds and MrgX2, and binding constant (K-A) of shikonin and acetylshikonin with MrgX2 were 2075.67 +/- 0.34 M-1 and 32201.36 +/- 0.35 M-1, respectively. Furthermore, beta-hexosaminidase and histamine release assay in vitro demonstrated that shikonin (1-5 mu M) and acetylshikonin (2.5-10 mu M) could both antagonize C48/80-induced allergic reaction. In conclusion, the MrgX2-SNAP-tag/CMC could be a reliable model for screening pseudo-allergy-related components from complex systems.
Keyword ：
Anti-pseudo-allergic Arnebiae Radix Cell membrane chromatography MrgX2 SNAP-tag
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| GB/T 7714 | Jia, Qianqian , Fu, Jia , Gao, Chunlei et al. MrgX2-SNAP-tag/cell membrane chromatography model coupled with liquid chromatography-mass spectrometry for anti-pseudo-allergic compound screening in Arnebiae Radix [J]. | ANALYTICAL AND BIOANALYTICAL CHEMISTRY , 2022 , 414 (19) : 5741-5753 . |
| MLA | Jia, Qianqian et al. "MrgX2-SNAP-tag/cell membrane chromatography model coupled with liquid chromatography-mass spectrometry for anti-pseudo-allergic compound screening in Arnebiae Radix" . | ANALYTICAL AND BIOANALYTICAL CHEMISTRY 414 . 19 (2022) : 5741-5753 . |
| APA | Jia, Qianqian , Fu, Jia , Gao, Chunlei , Wang, Hong , Wang, Saisai , Liang, Peida et al. MrgX2-SNAP-tag/cell membrane chromatography model coupled with liquid chromatography-mass spectrometry for anti-pseudo-allergic compound screening in Arnebiae Radix . | ANALYTICAL AND BIOANALYTICAL CHEMISTRY , 2022 , 414 (19) , 5741-5753 . |
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Abstract ：
Cell membrane chromatography is an effective method for screening bioactive components acting on specific receptors in complex systems, which maintains the biological activity of the membrane receptors and improves screening efficiency. However, traditional cell membrane chromatography suffers from poor stability, resulting in a limited life span and low reproducibility, greatly limiting the application of this method. To address this problem, cyanuric chloride-decorated silica gel was used for the covalent immobilization of the cell membranes. Cyanuric chloride reacts with amino groups on the cell membranes and membrane receptors to form covalent bonds. In this way, the cell membranes are not easy to fall off. The column life of the cyanuric chloride-decorated epidermal growth factor receptor/cell membrane chromatography column was extended to more than 8 days, whereas the column life of the normal cell membrane chromatography column dropped sharply in the first 3 days. A cyanuric chloride-decorated epidermal growth factor receptor/cell membrane chromatography online HPLC-IT-TOF-MSn system was applied for screening drug leads from Trifolium pratense L. One potential drug lead, formononetin, which acts on the epidermal growth factor receptor, was screened. Our strategy of covalently immobilizing cell membrane receptors also improved the stability of cell membrane chromatography.
Keyword ：
cell membrane chromatography epidermal growth factor receptor traditional Chinese medicine
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| GB/T 7714 | Fu, Jia , Jia, Qianqian , Liang, Peida et al. Enhanced stability designs of cell membrane chromatography for screening drug leads [J]. | JOURNAL OF SEPARATION SCIENCE , 2022 , 45 (14) : 2498-2507 . |
| MLA | Fu, Jia et al. "Enhanced stability designs of cell membrane chromatography for screening drug leads" . | JOURNAL OF SEPARATION SCIENCE 45 . 14 (2022) : 2498-2507 . |
| APA | Fu, Jia , Jia, Qianqian , Liang, Peida , Wang, Saisai , Zhou, Huaxin , Zhang, Liyang et al. Enhanced stability designs of cell membrane chromatography for screening drug leads . | JOURNAL OF SEPARATION SCIENCE , 2022 , 45 (14) , 2498-2507 . |
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Mast cells (MCs) are main effector cells in chronic spontaneous urticaria (CSU). Both Fc epsilon RI (FceRI)-and MAS-related G coupled receptor-X2 (MRGPRX2)-mediated MC activations affect CSU course. Leukocyte mono-immunoglobulin-like receptor 3 (CD300f) has been shown to regulate FceRI activation. However, no study has verified CD300f is a target to cure CSU. Therefore this study aimed to verify whether clarithromycin (CLA) regulates FceRI-and MRGPRX2-mediated MC activations via CD300f and shows therapeutic effect on CSU. The target of CLA was verification. CLA inhibited FceRI-and MRGPRX2-mediated MC activations were shown in vivo and in vitro. A single-center, self-comparison study was performed, and CLA-treated CSU was investigated in 28 patients who were not sensitive to the third-generation antihistamines. Serum inflammatory mediators in pa-tients before and after CLA administration were analyzed. CLA effectively inhibited type I anaphylactic reactions and pseudo-allergic reactions in mice. Moreover, CLA inhibited FceRI-and MRGPRX2-mediated MC signaling pathway activation. Regulatory effects of CLA were decreased significantly after CD300f knockdown. CLA effectively alleviated the symptoms of wheal and itch and reduced serum cytokine levels in patients. CLA negatively regulated FceRI-and MRGPRX2-mediated MC activation via CD300f and showed significant thera-peutic effect on CSU.
Keyword ：
Chronic spontaneous urticarial Clarithromycin Leukocyte mono-immunoglobulin-like receptor 3 Mast cell Negatively regulation
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| GB/T 7714 | Che, Delu , Zhang, Tao , Zhang, Tianxiao et al. Clarithromycin-treated chronic spontaneous urticaria with the negative regulation of FceRI and MRGPRX2 activation via CD300f [J]. | INTERNATIONAL IMMUNOPHARMACOLOGY , 2022 , 110 . |
| MLA | Che, Delu et al. "Clarithromycin-treated chronic spontaneous urticaria with the negative regulation of FceRI and MRGPRX2 activation via CD300f" . | INTERNATIONAL IMMUNOPHARMACOLOGY 110 (2022) . |
| APA | Che, Delu , Zhang, Tao , Zhang, Tianxiao , Zheng, Yi , Hou, Yajing , Geng, Songmei et al. Clarithromycin-treated chronic spontaneous urticaria with the negative regulation of FceRI and MRGPRX2 activation via CD300f . | INTERNATIONAL IMMUNOPHARMACOLOGY , 2022 , 110 . |
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Abstract ：
Since December 2019, the new coronavirus (also known as severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2, 2019-nCoV])-induced disease, COVID-19, has spread rapidly worldwide. Studies have reported that the traditional Chinese medicine Salvia miltiorrhiza possesses remarkable antiviral properties; however, the anti-coronaviral activity of its main components, salvianolic acid A (SAA), salvianolic acid B (SAB), and salvianolic acid C (SAC) is still debated. In this study, we used Cell Counting Kit-8 staining and flow cytometry to evaluate the toxicity of SAA, SAB, and SAC on ACE2 (angiotensin-converting enzyme 2) high-expressing HEK293T cells (ACE2(h) cells). We found that SAA, SAB, and SAC had a minor effect on the viability of ACE2(h) cells at concentrations below 100 mu M. We further evaluated the binding capacity of SAA, SAB, and SAC to ACE2 and the spike protein of 2019-nCoV using molecular docking and surface plasmon resonance. They could bind to the receptor-binding domain (RBD) of the 2019-nCoV with a binding constant (K-D) of (3.82 +/- 0.43) e-6 M, (5.15 +/- 0.64)e-7 M, and (2.19 +/- 0.14)e-6 M; and bind to ACE2 with K-D (4.08 +/- 0.61)e-7 M, (2.95 +/- 0.78)e-7 M, and (7.32 +/- 0.42)e-7 M, respectively. As a result, SAA, SAB, and SAC were determined to inhibit the entry of 2019-nCoV Spike pseudovirus with an EC50 of 11.31, 6.22, and 10.14 mu M on ACE2(h) cells, respectively. In conclusion, our study revealed that three Salvianolic acids can inhibit the entry of 2019-nCoV spike pseudovirus into ACE2(h) cells by binding to the RBD of the 2019-nCoV spike protein and ACE2 protein.
Keyword ：
2019‐ ACE2 nCoV salvianolic acid A salvianolic acid B salvianolic acid C
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| GB/T 7714 | Hu, Shiling , Wang, Jue , Zhang, Yongjing et al. Three salvianolic acids inhibit 2019-nCoV spike pseudovirus viropexis by binding to both its RBD and receptor ACE2 [J]. | JOURNAL OF MEDICAL VIROLOGY , 2021 , 93 (5) : 3143-3151 . |
| MLA | Hu, Shiling et al. "Three salvianolic acids inhibit 2019-nCoV spike pseudovirus viropexis by binding to both its RBD and receptor ACE2" . | JOURNAL OF MEDICAL VIROLOGY 93 . 5 (2021) : 3143-3151 . |
| APA | Hu, Shiling , Wang, Jue , Zhang, Yongjing , Bai, Haoyun , Wang, Cheng , Wang, Nan et al. Three salvianolic acids inhibit 2019-nCoV spike pseudovirus viropexis by binding to both its RBD and receptor ACE2 . | JOURNAL OF MEDICAL VIROLOGY , 2021 , 93 (5) , 3143-3151 . |
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Abstract ：
Anaphylactoid reactions are potentially fatal allergic diseases caused by mast cells (MCs), which release histamine and lipid mediators under certain stimuli. Therefore, there is an urgent need to develop new drug candidates to treat anaphylactoid reactions. The MrgX2 receptor mediates anaphylactoid reactions that cause inflammatory diseases. Cortex dictamni is a Chinese herb used for treating allergy-related diseases; however, its active compound is still unknown and its mechanism of action has not yet been reported. The aim of this study was to screen the anti-anaphylactoid compound from C. dictamni extracts. An MrgX2/CMC-HPLC method was established for screening MrgX2-specific compounds retained from the alcohol extract of C. dictamni. A mouse model of hindpaw extravasation was used to evaluate the anti-anaphylactoid effect of this ingredient. Intracellular Ca2+ mobilization was assessed using a calcium imaging assay. Enzyme immunoassays were performed to measure cytokine and chemokine release levels. The molecular signaling pathways were explored by western blotting. As a result, dictamnine was identified as an effective compound using the MrgX2/CMC method, which remarkably suppressed MC intracellular Ca2+ mobilization and the release of de novo degranulated substances, and inhibited PKC-PLC gamma-IP3R-associated protein signaling molecules. Hence, dictamnine is a novel therapeutic candidate for anaphylactoid reactions via MrgX2.
Keyword ：
anaphylactoid effects anti‐ CMC dictamnine mast cells MrgX2
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| GB/T 7714 | Liu, Rui , Hu, Shiling , Ding, Yuanyuan et al. Dictamnine is an effective anti-anaphylactoid compound acting via the MrgX2 receptor located on mast cells [J]. | PHYTOTHERAPY RESEARCH , 2021 , 35 (6) : 3181-3193 . |
| MLA | Liu, Rui et al. "Dictamnine is an effective anti-anaphylactoid compound acting via the MrgX2 receptor located on mast cells" . | PHYTOTHERAPY RESEARCH 35 . 6 (2021) : 3181-3193 . |
| APA | Liu, Rui , Hu, Shiling , Ding, Yuanyuan , Wang, Jue , Wang, Yuejin , Gao, Jiapan et al. Dictamnine is an effective anti-anaphylactoid compound acting via the MrgX2 receptor located on mast cells . | PHYTOTHERAPY RESEARCH , 2021 , 35 (6) , 3181-3193 . |
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In the present study, the oral bioavailability of cadmium (Cd) in earthworms and leeches was investigated through in vitro physiologically based extraction test (PBET) digestion/Caco2 and MDKC cell models. We are the first to create an innovative assessment strategy which has capacity to offer a more precise evaluation of Cd-associated health risks in traditional animal medicines (TAMs), by combinational usage of bioavailable Cd levels, the duration and frequency of the exposure to TAMs obtained by questionnaire data, as well as safety factor of TAMs. Our data showed that the percentage of bioavailability for Caco-2 cells in earthworms and leeches ranged from 3.29 to 14.17% and 4.32 to 12.61%, respectively. The percentage of bioavailability of MDCK cells in earthworms and leeches ranged from 4.83 to 15.74% and 6.53 to 15.04%, respectively. After adjusting by the bioavailability of Cd to target hazard quotient (THQ), excitingly, our findings manifested that the health risks induced by the ingestion of earthworms and leeches were acceptable in the clinic. Our key findings suggest that bioavailability characterization cannot be ruled out and health risks should be assessed on the basis of the bioavailable Cd levels rather than total levels. Our novel strategy provides insight into the bio-accumulation of Cd in organisms as well as a more realistic and accurate assessment of Cd-associated health risks in TAMs, with the main purpose of improving public health by scientifically using TAMs.
Keyword ：
Bioavailability Caco-2 cell model Cd In vitro digestion MDCK cell model Risk assessment Traditional animal medicines
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| GB/T 7714 | Zuo, Tian-tian , Luo, Fei-ya , He, Huai-zhen et al. Novel bioavailability-based risk assessment of Cd in earthworms and leeches utilizing in vitro digestion/Caco-2 and MDCK cells [J]. | ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH , 2021 , 29 (18) : 26513-26523 . |
| MLA | Zuo, Tian-tian et al. "Novel bioavailability-based risk assessment of Cd in earthworms and leeches utilizing in vitro digestion/Caco-2 and MDCK cells" . | ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH 29 . 18 (2021) : 26513-26523 . |
| APA | Zuo, Tian-tian , Luo, Fei-ya , He, Huai-zhen , Jin, Hong-yu , Sun, Lei , Xing, Shu-xia et al. Novel bioavailability-based risk assessment of Cd in earthworms and leeches utilizing in vitro digestion/Caco-2 and MDCK cells . | ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH , 2021 , 29 (18) , 26513-26523 . |
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Abstract ：
Background Allergic asthma is a common inflammatory lung disease and a major health problem worldwide. Mast cells (MCs) play a key role in the early-stage pathophysiology of allergic asthma. Substance P (SP) functions in neurogenic inflammation by activating MCs, and therefore, it may to participate in the occurrence and development of asthma. Objective We examined the relationship between SP and lung inflammation, and also whether SP can directly trigger asthma. Methods We measured the number of peripheral blood eosinophils, neutrophils and basophils and evaluated the levels of IgE and SP in blood samples of 86 individuals with allergic asthma. Serum IgE and SP levels were also determined in 29 healthy individuals. C57BL/6 mice were subjected to different doses of SP, and bronchoalveolar lavage fluid (BALF) was collected to count the inflammatory cells. Lung tissues were analysed using histopathological methods to evaluate lung peribronchial inflammation, fibrosis and glycogen deposition. Levels of IgE, interleukin (IL)-1, IL-2, IL-4, IL-5, IL-13, IL-17 and IFN-gamma were determined in mouse serum. Results Substance P levels were increased in the serum samples of patients with asthma. SP induced mouse lung peribronchial inflammation, fibrosis and glycogen deposition, with high levels of Th2-related cytokines such as IL-4, IL-5 and IL-13 observed in the BALF. Furthermore, low level of total IgE was noted in the serum, and SP had little effect on MC-deficient kit(W-sh/W-sh) mice. Conclusions & clinical relevance Substance P levels increased significantly in serum of asthmatic patients and independently associated with the risk of asthma. Furthermore, SP induced Th2 lung inflammation in mice, which was dependent on MCs.
Keyword ：
asthma mast cell SP Th2
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| GB/T 7714 | Wang, Nan , Wang, Jue , Zhang, Yongjing et al. Substance P-induced lung inflammation in mice is mast cell dependent [J]. | CLINICAL AND EXPERIMENTAL ALLERGY , 2021 , 52 (1) : 46-58 . |
| MLA | Wang, Nan et al. "Substance P-induced lung inflammation in mice is mast cell dependent" . | CLINICAL AND EXPERIMENTAL ALLERGY 52 . 1 (2021) : 46-58 . |
| APA | Wang, Nan , Wang, Jue , Zhang, Yongjing , Hu, Shiling , Zhang, Tianxiao , Wu, Yuanyuan et al. Substance P-induced lung inflammation in mice is mast cell dependent . | CLINICAL AND EXPERIMENTAL ALLERGY , 2021 , 52 (1) , 46-58 . |
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