The Rhei Radix et Rhizoma was one of the most widely used traditional Chinese medicine for its special biological activities. The content of rhein, one of its major compounds, was an important standard for the quantity control of Rhei Radix et Rhizoma. The major method used for the detection of rhein was instrumental analysis like HPLC, but it was complex, time-consuming and cannot detect large samples at the same time. The enzyme-linked imunmosorbent assay (ELISA) was accurate, reliable, simple, low costs, and of a high-throughout. Recently, it was widely used for the determination of those small molecule compounds in some traditional Chinese medicinal plants. In this study, an artificial antigen were synthesized by the carbodiimide (CDI) method. Rhein-bovine (rhein-BSA) conju gate and rhein-ovalbumin (rhein-OVA) conjugate, were produced as the immunogen and coating antigen, respectively. The conjugate and the hapten number in the conjugate were determined by UV-Vis spectrophotometry (UV). The conjugation ratio of Rhein and BSA was about 4.0:1, rhein acid and OVA was 2.6 : 1, respectively. Rhein-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The antiserum titer of the Rhein were higher than 8000 detected by ELISA. The successfully synthesized conjugate antigen rhein-BSA implies its feasibility in the establishment of fast immunoassay for the rhein content determination.
Animals ; Anthraquinones ; analysis ; immunology ; Antibodies ; analysis ; immunology ; Antigens, Plant ; analysis ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Mice ; Mice, Inbred BALB C ; Rheum ; chemistry ; immunology ; Rhizome ; chemistry ; immunology

This article mainly summarise the results of the chemical compositions and their pharmacological activities of Arnebiae Radix since 1966. The chemistry components isolated from Arnebiae Radix are mainly naphthoquinone, monoterpene phenol and quinone, phenolic acids and their salts, alkaloids, aliphatic and esters. Pharmacological results showed that the chemical compositions and the extracts of Arnebiae Radix have antibacterial, anti-inflammatory, anti-viral, hepatoprotection, antioxidant, anti-tumor and immune function and other activities. This article hopefully to provide a reference for further research, development and utilization of Arnebiae Radix.
Animals ; Boraginaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Plant Roots ; chemistry

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

A sensitive antibody-based lateral flow dipstick was developed for ginsenoside Re (GRe) detection. The stick consisted of a sample pad, a conjugate pad, membrane and an absorbent pad. The membrane was coated with two capture reagents, GRe-BSA conjugate and goat anti-mouse antibodies, forming a test line and a control line, respectively. The conjugate pad was saturated with colloidal gold particles coated with affinity purified monoclonal anti-GRe antibody. The visual detection limit was 200 microg x L(-1) of GRe and the reaction time was 10 min. The Panax ginseng roots were identified after these samples (10 mg) were extracted with 5 mL tap water for 30 min at room temperature, and the extracts were tested by the dipsticks and ELISA kit. The true and false P. ginseng could be distinguished with dipsticks. The dipstick could be used to detect the quality of the P. ginseng samples when the extract was diluted 100-folds. The results were compared with those obtained using an indirect competitive enzyme-linked immunosorbent assay (icELISA). The dipstick assay proved to be a sensitive and rapid tool for quality control of P. ginseng.
Animals ; Antibodies, Monoclonal ; immunology ; Counterfeit Drugs ; analysis ; Ginsenosides ; analysis ; Immunoassay ; instrumentation ; methods ; Mice ; Panax ; chemistry ; Reagent Strips ; Time Factors

Molecular Pharmacognosy is a new interdisciplinary subject formed by the organic integration of molecular biology and pharmacognosy. It is highly practical and innovative. In the course of teaching,both experimental teaching and theoretical teaching are of great significance. " Molecular Identification of Traditional Chinese Medicine" and the traditional teaching mode of confirmatory experiment are the preferred choices for the establishment of Molecular Pharmacognosy experimental courses in universities and colleges. Molecular Pharmacy is a forward-looking discipline with many emerging methods and technologies. Basic experimental teaching is not enough for students to learn this subject better,so it is especially important to introduce the latest scientific research results in experimental teaching. Experimental teaching based on the transformation of the latest scientific research results not only enables students to master basic experimental skills,but also broadens the breadth of students' knowledge,cultivates students' scientific research ideas,stimulates students' innovation spirit. Some suggestions and prospects have been put forward for the compilation of experimental teaching materials,the construction of experimental platform,the cultivation of teachers and academic exchanges. It is hoped that the contents of experimental textbooks will be developed from confirmatory experiments to comprehensive experiments,and the experimental platform for rational,standardized and efficient use will be built. Meanwhile,experimental courses involving multiple fields can be completed by multi-disciplinary teachers,and it is encouraged to actively carry out and participate in flexible,diverse,lively and interesting teaching practices. All the suggestions are intended to promote the development of Molecular Pharmacognosy.

The quality evaluation of traditional Chinese medicine can be divided into two aspects, effectiveness and safety. The existing methods for evaluating the quality of traditional Chinese medicine(TCM) are mostly based on the testing instruments, which can not meet the practical needs of simple, rapid and on-site in production and life. Immunoassay, characterized by simple, rapid, sensitive, specific, low-cost and high-throughput, is widely used in the fields of clinical diagnosis, environmental pollution monitoring, food safety testing, and other fields. In recent years, immunoassay technology has been gradually applied in the field of quality control of TCM, involving quantitative detection of effective components of TCM, detection of harmful substances in TCM, and detection of exogenous pollutants in TCM. This paper summarizes the principle of the wide application of ELISA and colloidal gold immunostrip technology and its application in quality evaluation of TCM, this technique has a good application prospect in the field of rapid detection of the quality of TCM. In this paper, the principles of the widely used ELISA and colloidal gold immune test strip technology as well as their application in quality evaluation of TCM were reviewed, and the results showed that this technique had a good application prospect in the field of rapid detection of the quality of TCM.

Endophytic bacteria exist in the host and do not cause obvious damage to the host，which is an important part of the host ecosystem. Endophytic bacteria are a kind of important microbial resources with diverse species and wide distribution. In the process of long-term coevolution between endophytic bacteria and host，many symbiotic relationships，such as antagonism and reciprocity，have been gradually formed，which can directly or indirectly regulate the growth and development of host，assist host to resist diseases and insect pests，environmental stress and other biological processes. Endophytic bacteria can produce a variety of secondary metabolites in the process of symbiosis with the host，which plays an important role in the development of new natural products. In recent years，endophytic bacteria and their secondary metabolites have been widely used in the research and development of new drugs，biological control and other fields. In this paper，the diversity，species identification and secondary metabolites of endophytic bacterial resources are reviewed，and the future development direction is discussed，hoping to provide reference for the further development and utilization of endophytic bacterial resources.

This study is to establish a pre-column derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) UPLC-MS/MS method for the determination of the monosaccharide composition of 12 polysaccharides. At the same time, the monosaccharide components of polysaccharides in Armillaria gallica were analyzed. The separation was performed on a ACQUITY ZORBAX RRHD Eclipse Plus C₁₈ column(2.1 mm×100 mm, 1.8 μm),using 95% acetonitrile (A) and ammonium acetate-5% acetonitrile-water (B) as mobile phase with gradient elution. The target components were detected in multiple-reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in ionization mode. The results showed that based on the monosaccharides detection method established by UPLC-MS/MS, the linearity of the 12 monosaccharides components were linear in their linear range (R²>0.990), and the recovery rate were 92.30%-105.6%. 11 monosaccharides such as fructose, mannose, and glucose were detected in A. gallica samples. The method established in this experiment is robust, highly reproducible and accurate, and is suitable for the determination of monosaccharide components such as A. gallica.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Monosaccharides ; Polysaccharides ; Tandem Mass Spectrometry

With the rice-steamed Rehmanniae Radix unearthed from the tomb of Haihunhou in the Western Han Dynasty as the re-ference, the present study evaluated the quality of Rehmanniae Radix and investigated the processing technology of rice-steamed Rehmanniae Radix to lay the foundation for the research on rice-steamed Rehmanniae Radix products. With catalpol and rehmannioside D as the investigation indexes, the quality and grade of Rehmanniae Radix from different producing areas were evaluated with the methods in 2020 edition of Chinese Pharmacopoeia. UPLC method was established for the determination of catalpol and rehmannioside D in the rice-steamed Rehmanniae Radix. The effects of steaming time, the amount of supplementary rice, and steaming times in the rice-steamed processing on the quality of products were investigated by L_9(3~4) orthogonal test and multi-index comprehensive balance scoring method combined with the content of catalpol and rehmannioside D and appearance characteristics. At last, the stability of the processing technology was tested. The results showed that the optimal processing technology for rice-steamed Rehmanniae Radix was as follows: Rehmanniae Radix and rice(200 g∶4 g) were steamed twice at atmospheric pressure, four hours each time. The mass fractions of catalpol and rehmannioside D were 0.184% and 0.335%, respectively, and the character score was 6.5. The processing conditions are reaso-nable, stable, and feasible. It can provide a basis for the restoration of the ancient rice-steamed processing technology and references for the development of rice-steamed Rehmanniae Radix products in the future.
Drugs, Chinese Herbal/pharmacology* ; Oryza ; Plant Extracts ; Rehmannia ; Technology

Objective:As a source of energy for Armillaria mellea and Gastrodia elata，the woods species as fungus material of G.elata are diverse and play an important role in the development of G.elata industry. In order to explore the impact of different woods species on the quality of G. elata，the plant origins and lignocellulose content of the woods，the yield and quality of G. elata per unit area were systemically analyzed through literature research and investigation on production bases. Method:G. elata and its cultivated woods were collected from four main producing areas （Guizhou, Yunnan, Hubei, and Shaanxi），and the the plant origins of the woods were identified by DNA fragments. The content of lignocellulose in the woods was determined by high-performance liquid chromatography and loss-on-ignition method. The content of polysaccharides of G. elata from these 4 areas was determined by Phenol-sulfuric acid method. The yields and polysaccharide content of G. elata cultivated with different woods species were compared and their correlation with the woods was analyzed. Result:The woods as fungus material of G. elata were diverse in species, and betulaceae was the most widely used species in cultivation of G. elata. There were differences in the composition ratio of lignocellulose in the woods. Nyssaceae had the highest cellulose content，Moraceae had the highest hemicellulose content and Rosaceae had the highest lignin content. Different woods species had certain effects on the yield and polysaccharide content of G. elata. The maximum yield of G. elata was 1 285.51 g and the lowest yield was 379.30 g. The average mass fraction of polysaccharide content was 241.1 mg·g^-1，with a range of 87.95-411.2 mg·g^-1. The yield and polysaccharide content of G. elata were highly positively correlated with the cellulose content of the woods, and highly negatively correlated with the lignin content. Conclusion:Different woods have a significant impact on the yield and quality of G. elata. Choosing the appropriate woods species will be beneficial to the absorption of nutrients for A. mellea and the yield increase of G. elata. This study can provide a scientific basis for the selection of woods species during the cultivation of G. elata.