2.Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S.choleraesuis
Zhen HUANG ; Shiqi XIA ; Daofeng LIU ; Chengwei LIU ; Weihua LAI
Chinese Journal of Analytical Chemistry 2017;45(2):217-223
Immunomagnetic separation (IMS) was coupled with fluorescent microspheres lateral flow assay (FM-LFA) for rapid detection of S.choleraesuis in this study.The target bacteria were firstly enriched from sample by immunomagnetic beads (IMBs),then eluted by heat treatment and detected by fluorescent microspheres lateral flow test strip.The IMBs was labeled with 30 μg/mg antibody,and the capture efficiency was greater than 90% against 102-106 CFU/mL of S.choleraesuis with great specificity.The immunofluorescent microspheres were prepared by coupling 300 μg of 11 D8-D4 monoclonal antibody with 1 mg of fluorescent microspheres at pH 6.Monoclonal antibody 5F11-B11 (2.0 mg/mL) and donkey anti-mouse IgG (1.0 mg/mL) were sprayed on nitrocellulose membrane as test line and control line,respectively.The FM-LFA based on IMS was used to detect S.choleraesuis in PBS and milk.The limits of detection in PBS buffer and milk were 1.5×105 CFU/mL and 7.6×105 CFU/mL respectively,which were 10 and 200 times lower than that of traditional fluorescent microspheres lateral flow assay,respectively.The results showed that the method,which could enrich S.choleraesuis in milk effectively,could avoid matrix interference and improve the detection sensitivity,thus had a good application prospect.
3.Study on Species Characteristics of Chinese Patent Medicines for Antitussive Effect
Zhang WANG ; Daofeng JIANG ; Yi ZHANG ; Xianli MENG ; Xianrong LAI ; Shasha LIU ; Huai YANG ; Yongfeng ZHENG ; Shaorong QIN ; Yuling QING ; Jing HUANG ; Tao PENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(1):6-10
This study was aimed to reveal the species characteristics of Chinese patent medicines for antitussive ef-fect and provide references for developing new drugs. This research targeted Chinese patent medicines for antitussive effect which were included in the Pharmacopoeia of the People's Republic of China and the New National Chinese Patent Medicines as well as those characterized by keywords such as cough cure, cough alleviating, antitussive effect, cough, persistent cough. The analysis was made on the species characteristics, such as the number of Chinese patent medicines for antitussive effect, license number, ethnomedicine patent medicines, drugs for children use, protection of varieties of traditional Chinese medicine, the number of drugs, the generic names of drug, and drug forms. The results showed that 684 Chinese patent medicines for antitussive effect collected in this research had ac-counted for 8.60% of the total 7 260 of Chinese patent medicines. A total of 7 450 license numbers were approved, and 33% of the Chinese patent medicines shares one license number. One Chinese patent medicine owns 16.6 li-cense numbers on average. Ethnomedicine patent medicines had 3 Tibetan prescriptions such as the Shiwuwei Chenxiang pill and 4 Mongolian prescriptions, such as the Siwei Tumuxiang powder. Drugs for children accounted for 14%, including 9 forms. The type of the generic names of drug reached 16 and most of them originate from abbrevia-tions of the main drug in prescription. The number of drugs in prescription ranges from 8 to 16. Chinese patent medicines for antitussive effect involved 16 forms, of which the proportion of the use of solid preparation was higher than the liquid preparation. It was concluded that Chinese patent medicines for antitussive effect were characterized by such advantages such as a variety of species, various forms, the reasonable number of drugs, considerable medicine retail market share and drug for children use which can meet the clinical needs, and meanwhile some prob-lems, such as a lack of criteria for the generic names of drug, the homogenization of fierce competition, and inade-quacy of ethnomedicine patent medicines.
4.Study on Prescription Characteristics of Chinese Patent Medicines for Antitussive Effect
Zhang WANG ; Daofeng JIANG ; Yi ZHANG ; Xianli MENG ; Xianrong LAI ; Yamei LI ; Jingzhu WANG ; Long WU ; Yan CHEN ; Yi TAN ; Xue LI ; Yuling QING ; Shaorong QIN ; Jing HUANG ; Tao PENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(8):1759-1765
This study was aimed to reveal characteristics of functions, main indications and applications of Chinese patent medicines in order to provide references for development of new Chinese patent medicines for antitussive ef-fect. Chinese patent medicines recorded in the Pharmacopoeia of the People's Republic of China and the New Na-tional Chinese Patent Medicines were taken as research subjects. Chinese patent medicines classified in the function and indication with key words of cough-relieving, cough-suppressing, cough and chronic cough were col-lected. Analysis was made on the prescription characteristics, such as functions, main indications and applications. The results showed that there were 684 Chinese patent medicines and 462 terms, which were used for a total of 1533 times. The frequently-used terms are antitussive effect with expectorant, antitussive effect, expectorant, heat-clearing and exterior-releasing, antiasthmatic, wind-expelling and antipyretic, antitussive effect with antiasthmatic, purging lung heat, relieving exterior syndrome and moistening the lungs. All 684 Chinese patent medicines were used 48 ma-jor treatments for a total of 1107 times. The main indications include acute and chronic trachitis / bronchitis, exoge-nous wind-heat type of common cold, exogenous wind-cold type of common cold, cough with abundance of phlegm due to phlegm-heat obstructing the lung, acute upper respiratory tract infection, asthma, acute and chronic pharyngi-tis, lung heat cough, and etc. All 684 Chinese patent medicines used a total of 537 kinds of herbs and chemical medicines, including 441 kinds of herbal medicine, 36 kinds of animal medicine, 25 kinds of mineral medicine and 35 kinds of chemical medicine. The top ten herbs in terms of using frequency are Glycyrrhizae radix et rhizoma, Platycodonis radix, Armeniacae semen amarum, Menthae haplocalycis herba, Citri reticulatae pericarpium, Ephedrae herba, Scutellariae radix, Perilla frutescens (L.) Britt, Pinelliae rhizoma, Fritillariae cirrhosae bulbus. Among them, 1/4 of Chinese patent medicines used extract. Five kinds of food or fruits were also used, which were orange, fresh gin-ger, pear, watermelon and dried mushroom. It was concluded that Chinese patent medicines for antitussive effect can be adopted in the treatment of various kinds of diseases, especially for the treatment of cough(heat cough in partic-ular), abundance of phlegm, asthma, pharyngitis, laryngitis, and etc. The treatment methods of clearing heat, dis-pelling wind and relieving exterior syndrome played an important role in the realization of antitussive effect. However, the expression of functions and main indications is far from standardized and clear. Drugs commonly used are herbal medicine plus animal medicine, mineral medicine and chemical medicine as well as food and fruit. The discovery of preparations with certain clinical curative effect from ethnomedicine and the using of their extracts as a major ingre-dient are one important way to develop new Chinese patent medicines with antitussive effect.
5.Pien Tze Huang alleviates LPS-stimulated liver Kupffer cell injury by regulating miR-155 expression
Luzhen QIU ; Zhaobin YANG ; Shaozhen HE ; Daofeng HUANG ; Xiaomei CHENG ; Huiping CHEN ; Hao XIA
Chinese Journal of Emergency Medicine 2024;33(4):536-541
Objective:To explore the mechanism by which Pien Tze Huang improves liver Kupffer cell damage induced by lipopolysaccharide (LPS) by regulating the expression of miR-155.Methods:LPS induced liver Kupffer cells to establish a cell injury model to simulate septic liver injury. RT-qPCR was used to detect the expression of miR-155 in damaged cells, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the inflammatory response and apoptosis of damaged cells. Then we treated LPS-induced Kupffer cells with Pien Tze Huang at different concentrations (0 mg/L, 5 mg/L, 10 mg/L and 15 mg/L), and detected the expression of miR-155 in the cells, the inflammatory response of the cells and Apoptosis rate. MiR-155 was silenced in the cell injury model, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the effect of miR-155 on inflammatory response and apoptosis of model cells. Overexpression of miR-155 in damaged cells treated with Pien Tze Huang was used to detect changes in cellular inflammatory response and apoptosis. Data are expressed in the form of mean ± standard deviation, and each group of data is analyzed using t test or one-way analysis of variance.Results:In the LPS-induced liver Kupffer cell injury model, the expression of miR-155 was significantly increased ( P<0.05), the expression levels of pro-inflammatory factors IL-6 and TNF-α were significantly increased, and the anti-inflammatory factor IL-10 was significantly increased. was inhibited ( P<0.05), and the cell apoptosis rate was significantly increased ( P<0.05). After Pien Tze Huang treatment, the expression of miR-155 in damaged liver cells was inhibited ( P<0.05), the levels of cellular inflammatory factors IL-6 and TNF-α were inhibited, and the expression of anti-inflammatory factor IL-10 was promoted ( P<0.05). Inhibit cell apoptosis ( P<0.05). Silencing miR-155 reduced the inflammatory response and apoptosis rate of cells ( P<0.05). Overexpression of miR-155 can reverse the effect of Pien Tze Huang on liver cell injury ( P<0.05). Conclusions:In the model of LPS-induced liver Kupffer cell injury, Pien Tze Huang can inhibite the inflammatory response and apoptosis of cells by inhibiting the expression of miR-155.