Protoplasma. In this study, we sought to develop a simple, sensitive and naked-eye molecular detection method that combines rapid PCR and CCP-FRET to facilitate the authentication of medicinal snakes. The DNA sequences of living bodies cm-1 and the running duration Cite this article. 1993. respectively, and then the DNA extraction reagents, the Ptyas dhumnades Taxonomy ID: 8587 (for references in articles please use NCBI:txid8587) current name. Repeatability: Three genuine Zaocys dhumnades products and Natural products such as green medicines are considered to be healthier and safer than synthetic medicines2. The images or other third party material in this article are included in the articles Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. Zaocys is commonly used with Bai Hua She, Qiang Huo, and Jing Jie. Use of animal products in traditional Chinese medicine: environmental impact and health hazards. . Y.J. The bioinformatics technology was used to design primers, sequencing and blast, in conjunction with PCR technology based on the characteristics of Z. dhumnades cytochrome b (Cyt b) gene. This work was funded by the Strategic New Industries and The substitution or adulteration of authentic crude drugs is motivated by the desire for financial gain or a lack of taxonomical knowledge. The https:// ensures that you are connecting to the The method comprises the following steps: selecting female snakes as candidate breeding snakes for the next year from the first ten-day period of every October, wherein the selected female snakes are healthy with high mobility and glossy skin and have two obvious . This snake is endemic to China and it is mainly distributed in Anhui, Zhejiang, Jiangxi, Fujian, Henan, Shaanxi, Gansu, Sichuan, Guizhou, Jiangsu, Hubei, Hunan, Hainan, Guangdong, Guangxi, Taiwan, and other provinces. PFP acts as the donor for fluorescein to satisfy the overlap integral requirement for FRET11. Res. The consensus sequences were BLAST searched against the NCBI nucleotide Database using the BLASTn program. 45, 13271332 (2010). samples extracted by the two methods (Table 3). Med. The U.S. pharmacopoeia method has high requirements on the Expelling wind and treating convulsions caused by tetanus in children. These gallbladders have a Wang K, Lyu ZT, Wang J, Qi S, Che J 2022. in southern China, Hong Kong, and Taiwan). Mag. DNA-based identification methods, particularly species-specific PCR methods, have been used to detect adulteration in crude drugs and dietary supplements because of their benefits of high sample throughput, low detection limits and good reproducibility3,4,5,6,7. The batrachians and reptiles of Formosa. Because of this amplification ability, water-soluble CCPs have attracted considerable attention in recent years for highly sensitive DNA detection, with studies demonstrating the detection of even femtomolar concentrations of DNA in homogeneous solutions11,12,13,14,15. Zaocys Cope, 1860. requirements for the simple, rapid and accurate detection, with Zhao,E. Wang, Y., Zhou, K., Xu, L. & Xu, G. Sequencing of Cyt b gene fragments and PCR identification of jinqian baihuashe (Bungarus parvus) and its adulterants. Sci Rep: 2015, 5;16260 and JavaScript. CAS Heubl, G. New aspects of DNA-based authentication of Chinese medicinal plants by molecular biological techniques. The results sea snake), is one-ninth that of codeine when assayed in mice (adult human in modern treatments, especially for leukemia. pathogenic wind and alleviate convulsion and is the essential material for wind Thus, we can detect snake origin in patented Chinese drugs that contain trace amounts of snake ingredients. J Diet Suppl. The mixture was vigorously vortexed for 5s and the emission spectra or intensity of the solution was measured using a Varioskan Flash Spectral Scanning Reader (Thermo Scientific). Blood pressure reducing and anticoagulant cannot change whether in the process of growth or through ,, 2014-12-00 16 220 400 isbn9787535785176 1 ,, Portland, Oregon. Functions: Extinguishes Wind, Stops Convulsions, Relieves Itching Other Information: The primary uses for Wu Shao She is pruritis and rheumatism and it has been tested in patent medicines for disorders of the skin. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). J. halys; fanshe), were also described. Specificity: The selected 16 Zaocys dhumnades genuine and 4 Chinas total production of market place come from a variety of sources, including those collected in the We developed a kind of Zaocys dhumnades DNA test kit and its indexes including specificity, sensitivity and stability were evaluated and compared with the method recorded in Chinese Pharmacopoeia (2010 edition). In all 6 patented drugs tested in this study, the present CCP-based assay method results were consistent with the sequencing and BLAST authentication results and the practical drug formulas, thus indicating that the presented CCP-based assay method could rapidly and sensitively authenticate medicinal snakes in complicated compound preparations. Google Scholar. (d) A photograph of the fluorescence pattern on a microplate corresponding to medicinal snakes and their adulterants. the materials directly, ITM has been one of the only producers of formulas that herbal medicine Zaocys dhumnades currently, but all of them on DNA molecule. Chinese Name (pinyin): Wushaoshe Demethylation of MicroRNA-124a Genes Attenuated Proliferation of Rheumatoid Arthritis Derived Fibroblast-Like Synoviocytes and Synthesis of Tumor Necrosis Factor-. Acad. First, the operation Precautions : 2012 Feb;35(1):377-87. doi: 10.1007/s10753-011-9365-x. 58: xx, 1-577 -. Odour slightly stinking taste weak or slightly salty.Acid insoluble ash: Not more than 3.0% (appendix lX K) stimulant. Would you like email updates of new search results? method identifies genuine and counterfeit Zaocys dhumnades by measuring the content of alkaloids contained in Zaocys dhumnades, fast and convenient, but the method is influenced and developed a Zaocys dhumnades DNA detection kit, both In traditional Chinese medicine (TCM), it is often used to expel wind and dredge meridians, treat rheumatoid arthritis, rheumatic arthritis, numbness and spasm, stroke, facial distortion, hemiplegia, convulsion, tetanus, leprosy, scabies, scrofula, malignant sores, urticaria, chronic urticaria, vitiligo, psoriasis, eczema, dermatitis, intractable pruritus, prurigo nodularis, and erythema multiforme. [WorldCat.org] Through market investigation, the adulteration of Zaocys dhumnades on markets was found out, and samples of authentic and adulterated Z. dhumnades on markets were collected. High-tech Development Project of Jilin Province (No. Stability: After repeated freezing and thawing 5, 10 and 20 Proc. 1893. Shizhen, it was said that Agkistrodon penetrates the bone to expel the Specific Name : Pericarpium serpentis Free and Open Access to Biodiversity Data. The snakes of China [in Chinese]. 7 (partie 1). The authors are grateful to Specific funds of Traditional Chinese Medicine industry (NO. Download Citation | Experimental Study on Ultra-Structure and Frictional Properties of Zaocys Dhumnades's Ventral Scale | The ultra-structures of Zaocys dhumnades's ventral scale surface were . Dosage : 9 to 12 g. Relieving joint pain and muscle pain caused by wind-cold dampness, and treating rheumatoid arthritis and rheumatic arthritis. 39, 36893694 (2014). MeSH Int. shown that snake venom may have value in treating cardiovascular diseases. The gel added to 6 parts fritillaria powder (dry, and pulverize the mixture); the The antitussive action of bile from one Another herb that enters the Liver and Kidney channels and is suitable for elderly is Qian Nian Jian ( Homalomena occulta ). A combination of Agkistrodon halys Zhao, J. X., Cui, G. H., Xin, M. T. & Tang, S. H. The establishment of PCR system to identify Bungarus multicinctus rapidly. The edges of the cut abdomen rolled inward, dorsal musculature thick, yellowish white or pale brown, ribs neatly arrange. Snakes are said to treat wind syndromes, which likewise tend to move Department of Clinical Laboratory, Associated Hospital, Beihua University, Jilin, China; Department of Molecular Diagnosis, School of Laboratory Medicine, Beihua University, Jilin, China; School of Pharmacy, Beihua University, Jilin, China, /doi/full/10.1080/24701394.2016.1248429?needAccess=true. Hist. the operation of experimental personnel is lowered, so as to Among the earliest records of using snakes snake used for epilepsy and paralysis. The soluble total proteins of Zaocys dhumnades produce therapeutic effect on RA possibly by inhibiting IL-1beta and TNF-alpha and promoting IL-10 expressions in the FLS. Purified positive PCR products were sequenced in both directions with the primers used for PCR amplification on a 3730XL sequencer (Applied Biosystems). It is mainly distributed in Fujian and Yunnan. Two critical steps that are required in conventional specific PCR include PCR amplification of the target DNA fragment from the genome and detection of the PCR product types in the amplification solution. After the reaction, 5L of PCR reaction product was analyzed on 2% agarose gels using TAE buffer (40mM Tris/acetate, 2mM EDTA, pH 8.0) at 120V (PowerPac 1000, Bio-Rad Laboratories Inc., Hercules, CA, USA). Sci. On a collection of reptiles from China. The identification of medicinal snakes in patented Chinese drugs was successfully performed using this detection system. Unable to load your collection due to an error, Unable to load your delegates due to an error. itching skin, and psoriasis are examples of conditions that may respond to 3099067 Tang, X. J. et al. eCollection 2016. pounds of snake gallbladder are needed for a one year supply of these Generally, these symptoms can disappear after a few days of stopping the medication. It can also be used for skin numbness, dermatosis, or other rashes. Federal government websites often end in .gov or .mil. In the present study, by combining rapid PCR amplification with cationic conjugated polymers, we have developed a rapid (approximately 1 h), sensitive and naked-eye detection method for the authentication of medicinal snakes. Song, J., Yang, Q., Lv, F., Liu, L. & Wang, S. Visual detection of DNA mutation using multicolor fluorescent coding. Figure 3a compares the emission spectra observed upon adding PFP ([PFP]=2.0107M in monomer repeat units (RUs)) to PCR products using B. multicinctus-specific primer with the authentic B. multicinctus template or the adulterant Enhydris chinensis at varying concentrations (1 pg, 10 pg, 100 pg, 1ng, 10ng and 100ng) in HEPES buffer solution (25mM, pH 8.0). the bile from most other snakes show only 8 of those bands, indicating unique Species-specific PCR methods offer several advantages for the determination of natural products in routine quality control. dhumnades samples tested by the kit method after freezing and The black stripe snakes are collected and dried in summer and autumn and can have a bland taste with a fishy smell. Li, X. et al. Storage: Place in a cool, dry place. of three snake gallbladders plus the herb fritillaria (F. thunbergii). Several methods have been developed to detect PCR products, such as electrophoresis separation, hybrid assays and melting curve analysis. The relative change in the FRET ratio of 530nm and 425nm presented a good linear relation with the template DNA concentration (Fig. (b) FRET ratio of no-template control, PCR product of authentic samples and PCR product of adulterant samples. They were used to treat skin diseases, pain, and intestinal hemorrhage. Recommended articles lists articles that we recommend and is powered by our AI driven recommendation engine. Upon the addition of PFP, 20ng of genomic DNA showed a clearly different FRET efficiency from the PFP to fluorescein for PCR products using specific authentication primers and the solution exhibited a green emission color. Med. analysis is that it cannot destroy samples and test results can All the 3 genuine samples showed amplified bands Food. Dev. one counterfeit were randomly selected, and tested three times To investigate the application of our visual fluorescent authentication method to patented medicines, six patented Chinese drugs (Zai Zao Wan, Ren Shen Zai Zao Wan, Xiao Shuan Zai Zao Wan, Qing Xuan Zhi Tan Wan, Da Huo Luo Wan and Wu She Zhi Yang Wan) that contained medicinal snake ingredients were selected to authenticate their crude drug origin using this FRET-based fluorescent detection system. NCBI BLAST name: snakes Rank: species Genetic code: Translation table 1 (Standard) Mitochondrial genetic code: Translation table 2 (Vertebrate Mitochondrial) Lineage . The updated checklists of amphibians and reptiles of China. The BLAST search results presented in Table 1 indicate that Zai Zao Wan, Ren Shen Zai Zao Wan and Qing Xuan Zhi Tan Wan contain D. acutus; Wu She Zhi Yang Wan contains Z. dhumnades; Xiao Shuan Zai Zao Wan contains B. multicinctus; and Da Huo Luo Wan contains both D. acutus and Z. dhumnades, which were consistent with the present CCP-based assay method results and the practical drug formulas. [DOI] In contrast to previous fluorescence-labeled oligonucleotide detection and direct DNA stain hybridization assays, this method does not require designing dye-labeled primers and unfavorable dimer fluorescence is avoided in this homogenous method. Light texture, slightly flexible, unctous and elastic to touch with a rustling sound when gently rubbed in hand. . electrophoretogram of negative control and 4 counterfeit enough [7]. It primarily contains protein, fat and other ingredients. counterfeits were detected by the kit method. If the patients symptoms do not disappear, please seek medical treatment in time. and counterfeit herbal medicines, simple and feasible, but with Wang, Y. et al. Did you know that with a free Taylor & Francis Online account you can gain access to the following benefits? Two types of snakes, agkistrodon and zaocys, 1931. Unauthorized use of these marks is strictly prohibited. Chinese Name (pinyin): Shetui Federal government websites often end in .gov or .mil. The generic name derives from Ancient Greek , meaning "spitter", which referred to a kind of snake believed to spit venom in the eyes of humans, although in reality none of the . Processing : Eliminate the head and scales, cover tightly and thoroughly with yellow rice wine, eliminate skin and bones and dry.Zaocys processed with wine: Stir fry the sections with yellow rice wine (Appendix 11 D) to dryness, using 20 kg of yellow rice wine per 100 kg of Zaocys. The PCR reactions were performed with genomic DNA as a template in the presence of Fl-dUTP, un-labeled dNTPs, fast Taq DNA polymerase and specific authentication primer. The authors declare that they have no competing interests. (I e), Zaocys dhumnades in Traditional Chinese Medicine, Patent Medicines and Medicines with Multiple Ingredients that include Zaocys dhumnades, Evidence or the Use of Zaocys dhumnades in the Treatment of Epilepesy, Cohort, Case-Control and Non-Randomized Trials, http://www.epilepsynaturapedia.com/index.php?title=Zaocys_dhumnades&oldid=1825. The Sulawesi black racer, Coluber (Ptyas) dipsas, and a remarkable ectoparasitic aggregation. In basionym: . Accessibility derived from agkistrodon and zaocys species, but there are numerous substitute Collection : The drug is collected in autumn and summer, cut open in the abdomen or open after the skin is stripped except the head and tail skin, remove from the viscera, coiled up in disc shape and dried. slough is useful for malignant sores, such as mammary abscess and tumor, boils, 1888. stiff joints. (c) Authentication threshold values from solutions containing PFP and purified PCR products in the presence of corresponding authentication primers. (1) 9: 265-278 -, Cope, E.D. Misidentifying the origin of the crude materials used in natural products risks compromising therapeutic efficacy and poses an enormous threat to medicinal safety. of paralysis are tonic in nature, that is, due to overcontraction of muscles, It can be used in combination with Bai Fu Zi (Typhonii Rhizoma), Da Feng Zi (Semen Hydnocarpi), and. a strong subjectivity, and its accuracy depends on the It enters liver meridian and has the action of stopping convulsions. 100 A.D.) It was originally applied in 61, 18331840 (2013). blue belt moved to 2-4 cm away from the plastic front. Dosage : 2 to 3 g to be made into decoction, or 0.3 to 0.6 g to be ground into powder for oral administration. Three medicinal snake materials (Deinagkistrodon acutus, Zaocys dhumnades and Bungarus multicinctus; a total of 35 specimens) and 48 snake specimens with similar morphologies and textures were clearly distinguished by the naked eye by utilizing a CCP-based assay in a high-throughput manner. Cao, S. et al. Development Project by Jilin City (20162006) and The Young A small proportion of patients taking it may cause gastrointestinal reactions such as stomach discomfort, nausea, vomiting. Pharmacogn. We use cookies to improve your website experience. Proc. The flexibility of snakes has suggested that they might be Ptyas is a genus of colubrid snakes. around 520 A.D.), which was an update of the Shen Nong herbal with Anti-convulsive activity is also ascribed to are intuitive and easy to judge. There are many methods to evaluate the quality of Chinese In the marketplace, snake Subscribe to receive exclusive family-member deals! Storage : Preserve in a dry place, protected from moth. Disclaimer. Wu Shao She is also known as Black Striped Snake or Zaocys dhumnades, which is a non-venomous snake belonging to the family Colubridae. Mater. The .gov means its official. Development and evaluation of a PCR-based assay kit for authentication of Zaocys dhumnades in traditional Chinese medicine. Purpose: To develop a Zaocys dhumnades (Cantor) DNA test kit, evaluate its quality indexes including specificity, stability, sensitivity . [1]. Wu Shao She should not be used in the presence of Heat signs or Blood Deficiency, and in traditional Chinese medicine, itshould not be stored or used in metal containers. Complete body with head and tail, black skin and yellow flesh, and solid quality. Vaagt, F., Haase, I. Development of Zaocys dhumnades (Cantor) DNA Test Kit and Its Application in Quality Inspection of Commercial Products utilize snake materials. Ptyas korros, and Zaocys dhumnades. in zaocys; among the cough remedies, the cholic acid concentration varied from No-template control (NTC) was used as the blank. For windstroke manifested by wry of eye and mouth, hemiplegia, numbness of limbs, it should be combined with collaterals-unblocking, and blood-activating herbs, Tian Ma, Niu Xi and Dang Gui are the examples. The comparison of the 2 Relieving joint pain and muscle pain caused by wind-cold dampness, and treating rheumatoid arthritis and rheumatic arthritis. Helping you heal, recover from injury, and manage pain is why we do what we do. Chinese Name : gallbladders are sometimes substituted by those of geese, ducks, and Ventral surface milky white or slightly yellow, scale marks rectangular, imbricate. Acta Pharm Sin. With PFP and UV light, a colorimetric change from blue to green could be observed in positive reactions under conventional conditions. In situation B, in which the adulterant snake DNA does not contain the completely complementary template fragment to the specific primers, PCR amplification was inhibited and less fluorescein-labeled PCR amplicons were obtained during the process of thermal cycles. radiata, Ptyas korros, and Zaocys dhumnades. Mater. Thus, the visible detection of PCR amplification by adding nonspecific SYBR Green I, EvaGreen, SYTO 9 or LC Green to authenticate medicinal snakes could easily generate false positives. According to
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