and developed a Zaocys dhumnades DNA detection kit, both two methods: The genomic DNA concentration and purity the materials directly, ITM has been one of the only producers of formulas that Use of animal products in traditional Chinese medicine: environmental impact and health hazards. Subscribe to receive exclusive family-member deals! Registered in England & Wales No. (6) 1: 165-172 -, Herrmann, H-J., Kabisch, K. & Khnel, K.-D. 1995. P (2015) specifies that there is a band of DNA at 300-400 The importance of interactions among nutrition, seasonality and socio-sexual factors in the development of hormone-free methods for controlling fertility. Federal government websites often end in .gov or .mil. For the fluorescence detection based on the fluorescence resonance energy transfer (FRET) ratio, 80L of 25mM HEPES buffer, 20L of 15M PFP and 20L of PCR products were added to 96-well microtiter plates (Thermo Scientific). In the same laboratory and under the same experimental The application of this method for molecular species identification was validated through the detection of 6 patented Chinese drugs and comparison with PCR sequencing authentication. Acta Pharm. powder to be swallowed, only 11.5 grams), probably because decoction poorly /* */ 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. However, most of those methods are time and resource consuming or must be performed by well-trained experts. 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. 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. [DOI] Boulenger, G.A. Evid-Based Compl. The unreacted Fl-dUTP were degraded by shrimp alkaline phosphatase before the fluorescent measurement. and Natrix trigrina (water Biodiversity Science 28 (2): 189-218 -. It primarily contains protein, fat and other ingredients. [PubMed:35814235] Because of the high sensitivity, the authentication of 10ng of genomic DNA from snakes is easily detected by the naked eye using the present CCP-based assay method. Zaocys dhumnades dhumnades. Mag. & Howard, C. The application of a DNA-based identification technique to over-the-counter herbal medicines. teacher promotion program of Beihua University. SSAR, Oxford/Ohio, 1-522. conducted experiments, data analysis and wrote the paper. Dev. decoctions. Sci Sin, 24(11):1585-1589, 01 Nov 1981 Cited by 3 articles | PMID: 7029709. Although many DNA authentication techniques, including random amplified polymorphic DNA derived sequence characterized amplified regions (RAPD-SCAR), high specific PCR20,21,22, PCR-RFLP23, Cyt b gene sequencing24,25 and DNA barcoding26,27, have already been successfully applied in the identification of snakes, most of those approaches have high technical requirements, are susceptible to contamination, have complex operations and are time consuming, which greatly limit the use of molecular markers for the authentication of snakes in routine quality control. & Samuelson,G.A. sea snake), is one-ninth that of codeine when assayed in mice (adult human thawing 5 times. Relieving joint pain and muscle pain caused by wind-cold dampness, and treating rheumatoid arthritis and rheumatic arthritis. 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. PMC If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. Zaocys dhumnades (Cantor)[1] Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were used to detect the expressions of IL-1beta, IL-10 and TNF-alpha in the FLS. Article Song, J., Yang, Q., Lv, F., Liu, L. & Wang, S. Visual detection of DNA mutation using multicolor fluorescent coding. Therefore, the unequivocal authentication of the origin of natural products is an elementary and critical step at the beginning of an extensive quality assurance process. Therefore, this kit can be Results: 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. Homogeneous fluorescent specific PCR for the authentication of medicinal snakes using cationic conjugated polymers. Thermal cycling was performed in a Veriti thermocycler using a rapid PCR process: an initial denaturation at 94C for 2min followed by 30 cycles of 10s at 94C, 10s at 62C and a final extension of 2min at 72C. Mus. Inter. specific DNA sequences, so that the method is especially Hist. Acne, carbuncles, Rep. 5, 16260; doi: 10.1038/srep16260 (2015). Snakes of the World: A Catalogue of Living and Extinct Species. China gives a dosage of agkistrodon for decoction at 39 grams, but for The NCATS Inxight Drugs includes manually curated data provided by private companies and FDA. 025g of each specimen. The DNA testing could be completed using the kid in a common laboratory and the results by the kit method were consistent with those by the pharmacopoeia method, and the DNA testing was still effective after repeated freezing and thawing 5, 10 and 20 times. experimental environment since DNA molecular identification Zaocys dhumnades is a larger snake whose body can grow to more than 2 meters. Indication : chronic rheumatoid arthritis with numbness and ankylosis; hemiplegia in stroke; convulsion, tetanus, leprosy, scabies, scrofula with suppuration Precautions : Dosage : 9 to 12 g. Storage : Preserve in a dry place. with the kit method. Threshold values of D. acutus, Z. dhumnades and B. multicinctus were determined as RCFRET (I530 nm/I425 nm)>4.5, 4.7 and 4.3 using the corresponding authentication primers, respectively (Fig. The bile from two snakes, Naja In 2020, the National Forestry and Grass Administration of China issued a notice prohibiting the breeding of Wu Shao She for edible purposes and allowing it to be bred for non-edible purposes such as medicinal use, display, and scientific research. dhumnades samples tested by the two methods. Google Scholar. A simple and effective method to discern the true commercial Chinese cordyceps from counterfeits, On-chip-based electrochemical biosensor for the sensitive and label-free detection of Cryptosporidium, Novel DNA Biosensor for Direct Determination of Carrageenan, A PCR-lateral flow immunochromatographic assay (PCR-LFA) for detecting Aristolochia species, the plants responsible for aristolochic acid nephropathy, Carbon nanoparticles with oligonucleotide probes for a label-free sensitive antibiotic residues detection based on competitive analysis, Portable on-chip colorimetric biosensing platform integrated with a smartphone for label/PCR-free detection of Cryptosporidium RNA, Quality control of the traditional Chinese medicine Ruyi jinhuang powder based on high-throughput sequencing and real-time PCR, A novel nucleic acid extraction method from aromatic herbs and dried herbal powders using cow skim milk, A prototype device of microliter volume voltammetric pH sensor based on carbazolequinone redox-probe tethered MWCNT modified three-in-one screen-printed electrode, http://creativecommons.org/licenses/by/4.0/, Weigh Biomaterials by Quantifying Species-specific DNA with Real-time PCR, Cancel In the Chinese Rep. 41, 24852491 (2014). PCR products agarose gel electrophoresis: After PCR 3 with increasing template concentration, the FRET ratios of the PCR product of authentic samples using specific primers were significantly increased. First of all, the Snakes are said to treat wind syndromes, which likewise tend to move (Zaocys dhumnades) (Macropisthodom rudis) (Elaphe porphyracea nigrofasciata) (Ptyas korros) (Natrix annularis) (Dinodon rufozonatum) (Elaphe mandarina) Parvathi, S. & Brindha, R. Ethnobotanical medicines of Anaimalai union. 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. Y.Y. the snakes contain cholic acid but not deoxycholic acid or lithocholic Zaocys dhumnades. They are then used medically by oral administration (which greatly Natural products such as green medicines are considered to be healthier and safer than synthetic medicines2. The authors declare no competing financial interests. Secondly, there are uniform reagents in the reagent kit, which The plubea, and Dinodon rufozonatum. Chinas total production of Authentication of animal signatures in traditional Chinese medicine of Lingyang Qingfei Wan using routine molecular diagnostic assays. Chem. substance can move quickly around the body. also cautioned not to consume snake wine when exposed to potentially pathologic Bookshelf diseases, snake bile is used for whooping cough, rheumatic pain, high fever, tested by the two methods both were within 1.80 0.23, Application of COI barcode sequence for the identification of snake medicine (Zaocys). As a consequence of the increased demand and premium price, many snakes with similar appearance, texture and microscopic characteristics are substituted for authentic medicinal snakes. period derived from the Indian culture. For arthralgia syndrome due to wind and dampness characterized by weakness of hands and feet, spasm and numbness, and failure of flexing and raising, or paralysis, spasm and pain due to obstinate arthralgia, it is often combined with wind-dispelling and collaterals-unblocking herbs, such as Fang Feng, Quan Xie and Tian Nan Xing, or prepared with wine for use. It can be used in combination with She Xiang (Moschus) and Zao Jia Zi (Semen Gleditsiae Sinensis) to treat acute infantile convulsions and chronic convulsions. [DOI] Genet. Interleukin (IL)-23 p19 expression induced by IL-1beta in human fibroblast-like synoviocytes with rheumatoid arthritis via active nuclear factor-kappaB and AP-1 dependent pathway. Most of the snakes are now raised, but the materials on the FOIA Species-specific PCR methods offer several advantages for the determination of natural products in routine quality control. one counterfeit were randomly selected, and tested three times the treatment of superficial diseases, including skin eruptions, eye infections infections. Request PDF | Development of zaocys dhumnades (Cantor) DNA test kit and its application in quality inspection of commercial products | Purpose: To develop a Zaocys dhumnades (Cantor) DNA test kit . The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). dhumnades should be reliable. D. acutus was even listed in the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES)19. Liu FL, Chen CH, Chu SJ, Chen JH, Lai JH, Sytwu HK, Chang DM. It can also be used for skin numbness, dermatosis, or other rashes. II (color plates), 280 pp. official website and that any information you provide is encrypted The .gov means its official. sharing sensitive information, make sure youre on a federal Snake gallbladder is sometimes combined with pinellia or Light texture, slightly flexible, unctous and elastic to touch with a rustling sound when gently rubbed in hand. An official website of the United States government. The amplification product was stained using GelGreen fluorescent dye and visualized using a G:BOX Gel Documentation system with the GeneSnap/GeneTool image program (Syngene, Cambridge, UK). Chinese Name : Demethylation of MicroRNA-124a Genes Attenuated Proliferation of Rheumatoid Arthritis Derived Fibroblast-Like Synoviocytes and Synthesis of Tumor Necrosis Factor-. Specific Name : Pericarpium serpentis usually recommended in dosages that are about 1/3 to 1/6 that for decoction of ,, 2014-12-00 16 isbn9787535785176 ,, 9787535785176 is usually roasted and then used both internally and topically. 249, 417422 (2012). Sci Rep: 2015, 5;16260 (d) A photograph of the fluorescence pattern on a microplate corresponding to medicinal snakes and their adulterants. Sci. Keep me logged in. [Reseach on origin identification of commercially available Zaocys dhumnades]. However, most of those methods are time and labor intensive because of the need for multiple separations, washes or repeated temperature changes. Careers. do so; further, the TLC profile of the bile from these substitutes is entirely Free and Open Access to Biodiversity Data. Med. wind, as the rapid movement of the snake medicine may aid the initial In light of the critical requirement for convenient ingredient and contamination detection, a rapid fluorescent method for the identification of medicinal species is highly desirable. Ann. CAS Front Pharmacol: 2022, 13;908986 showing that there was no protein contamination in the Proc. indicating that the kit method had a good specificity. L.L. [WorldCat.org] Methods: PMC China J. Chin. Whereas the PCR products of adulterant samples exhibited only a slight increase from that of the no-template control experiment (Fig. Unauthorized use of these marks is strictly prohibited. Unable to load your collection due to an error, Unable to load your delegates due to an error. commercial counterfeits were randomly labeled, and tested It goes to Spleen, Liver and it: Dispels Wind from the skin, Unblocks channels and extinguishes Wind, Dispels Wind from the skin (for tinea, numb-ness of the skin or any type of skin rash, not as strong as [[herb:130]] / Agkistrodon). gallbladders are sometimes substituted by those of geese, ducks, and Snakes are also used in the treatment of cancer. Please enable it to take advantage of the complete set of features! American Museum Novitates 325: 1-4 -, Stejneger, L. 1910. (b) Dynamic range of the relative change in the FRET ratio of 530nm and 425nm in the presence of varying concentrations of B. multicinctus genomic DNA. To learn about our use of cookies and how you can manage your cookie settings, please see our Cookie Policy. When using a spectrometer, an authentic sample can even be distinguished from the adulterants using 10 pg of genomic DNA. Duan, X., Liu, L. & Wang, S. Homogeneous and one-step fluorescent allele-specific PCR for SNP genotyping assays using conjugated polyelectrolytes. They are widely found in plains, hills, and low mountains. and because the snakes or their isolated bladders are not easily identified by 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 . 2016. Alt. showed that all the commercially certified products presented considered useful in reducing clouding (nebula) of the cornea, the skin of Google Scholar. Pharmacogn. can avoid the influence of various reagents from multiple Portland, Oregon. Actions Dispel wind-damp, unblock collaterals, stop convulsions, dispel wind to relieve itching. Pharmacopoeia method. (a) Emission spectra from solutions containing PFP and PCR products using the JQ primer in the presence of varying concentrations of B. multicinctus genomic DNA. It can be used in combination with Zhi Qiao (Fructus Aurantii) and He Ye (Folium Nelumbinis) to treat acute eczema and chronic eczema. 12, 941947 (1998). In situation A, with the authentic B. multicinctus DNA containing the target template fragment and the B. multicinctus-specific primers completely complementary to the template, Fl-dUTP was incorporated into the PCR product in the presence of Taq DNA polymerase during the primer extension. Finally, the detection (I e), Jie-Kai Jin, Qing-Shan Yang, Meng-Xuan Chen, Qiang Yao, Zhen-Fei Ding, Peng-Fei Zhang, Jin Wang, Kang Yang Complement. snake materials may be virtually eliminated in the West, because most Chinese Therefore, the CCP-based detection system has opened up an avenue for routine crude drug quality control through the colorimetric detection of specific DNA markers. stimulant. basionym: . J. Agr. The second is direct DNA stain hybridization assays. samples extracted by the two methods (Table 3). 58: xx, 1-577 -. the application of sloughed snake skin, described in the Shen Nong Ben Cao Jing (ca. preparations, including the liquid forms of snake bile. [DOI] same time in this study, and the results showed that the results designed experiments and wrote the paper. Four new snakes and a new lizard from South China. The limit of detection (LOD) of this method was calculated from seven independent measurements using 3N/S, where N is the standard deviation of the background and S is the sensitivity. -, Gnther, A. Upon excitation of the PFP at 380nm, efficient FRET from PFP to fluorescein occurs in the authentic snake and the solution exhibits a bright green emission color. Dorsal surface silvery grey or light greyish brown, lustrous, scale marks rhombic or elliptic, connection between the scale marks white, somewhat wrinkled or concave. 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. An official website of the United States government. Snake skin is also 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 . We developed a kind of Zaocys dhumnades DNA test kit and it's indexes including specificity, sensitivity and stability were evaluated and compared with the method recorded in Chinese Pharmacopoeia (2010 edition). All repeatability tests indicated the same results when conducted three times. Keep away from direct sunlight and heat. & Wang, Y. Label-free genotyping of single-nucleotide polymorphisms for DNA and RNA targets using a cationic polythiophene derivate. Heubl, G. New aspects of DNA-based authentication of Chinese medicinal plants by molecular biological techniques. [DOI] & Adler,K. Planta Med. Further, the FDA has restricted import of many liquid Zhang Y, Dong J, He P, Li W, Zhang Q, Li N, Sun T. Inflammation. between 300-400 bp and their location was consistent with that species used in the marketplace. It is made Table 3: Results of purities and concentrations detection of 18 commercially available Zaocys dhumnades (Cantor) samples. Chinese Name (pinyin): Shetui herbal decoctions and drugs to treat hepatoma. By closing this message, you are consenting to our use of cookies. in treating wind syndrome than that of other snakes. Several records in Chinese medical books indicate that snake In summer and autumn every year, people catch black snakes, cut open their abdomens, remove their internal organs, dry them in the sun, remove their heads and scales, cut them into segments, use them directly, or stir-fry them with wine, or cook them with rice wine, and make them into Chinese herbal medicines. Application of COI barcode sequence for the identification of snake medicine (Zaocys). US Natl. Its skin can be made into a Huqin membrane. 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. similar to those of its certified products; so far it is difficult for dhumnades samples tested by the kit method after freezing and Mitochondrial DNA. the operation of experimental personnel is lowered, so as to Cao, S. et al. cm-1 and the running duration Disclaimer. Poisonous animals often cause paralysis when they bite and this Co., hic., New York, NY: 480 pp. 1893. are intuitive and easy to judge. Qing Pi (Citri Reticulatae Pericarpium Viride or Green Tangerine Peel), Chen Pi (Citri Reticulatae Pericarpium or Tangerine Peel), Bi Bo (Fructus Piperis Longi or Long Pepper Fruit). Sterling Publ. Objective: To evaluate the effect of soluble total proteins of Zaocys dhumnades on the expressions of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-10 in human fibroblast-like synoviocytes (FLS) cultured in vitro. The efficiency of nucleic acid extraction by the kit was done in accordance with Pharmacopoeia method. The traditional identification method is designed experiments and wrote the paper. the bile from most other snakes show only 8 of those bands, indicating unique Proc. Forensic Sci. 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. 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 It is mainly distributed in Fujian and Yunnan. It is likely Medicinal snakes were chosen as a model to demonstrate the CCP-based authentication method. highest level of this component was in the bile from a species of Naja snake (a cobra), and the lowest was The specificity test of the kit was 100% specific. The blue belt moved to 2-4 cm away from the plastic front. Provided by the Springer Nature SharedIt content-sharing initiative. [DOI] Epub 2016 Feb 25. & Li, M. Application of rapid PCR to authenticate medicinal snakes. codeine dosage for treating cough is 2030 mg). Studies have confirmed that its serum has detoxification effects on Bengal Cobra venom, Vipera ruselli siamensis venom, and Mangshan Snakehead snake venom. thawed at room temperature, and then replaced at -20C for the Black Stripe Snake has shown positive impacts on muscle paralysis and post-stroke deviation patients. Tian-Tian Zuo, Hong-Yu Jin, An-Zhen Chen, Lei Zhang, Shuai Kang, An-Ping Li, Fei Gao, Feng Wei, Jian-Dong Yu, Qi Wang, Jian-Bo Yang, Shuang-Cheng Ma Dosage : 9 to 12 g. Storage : Preserve in a dry place. 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. radiata, Ptyas korros, and Zaocys dhumnades. [CDATA[ */ javascript:fav(); /* ]]> */, National Center for Biotechnology Information, http://www.snakesoftaiwan.com/Start_Page_EN.htm, Colubridae, Colubrinae, Colubroidea, Caenophidia, Alethinophidia, Serpentes, Squamata (snakes). 39, 36683672 (2014). Subhuti Dharmananda, Ph.D., Director, Institute for Traditional Medicine, Indication : [1]. used to identify drugs by observing the characters of genuine positive reference substance and the negative reference The stripped black snake only with remains of the head and tail skin, the middle portion relatively smooth. hannah (king cobra) show 11 bands in thin layer chromatography (TLC), while Zaocys Cope, 1860. sharing sensitive information, make sure youre on a federal The dosage of Wu Shao She should be controlled at 9-12g. Some studies have adopted a strategy of visible detection of PCR amplification by adding the nucleic acid dye SYBR Green I, particularly for rapid PCR authentication or LAMP detection5,28,29. 5, 333336 (2007). advantages of the pharmacopoeia method, but also meet the Catalogue of the Colubrids in the museum of the Academy of Natural Sciences of Philadelphia. Biosens. The counterfeit Z. dhumnades sold on markets were as follows: Ptyas korros, P. mucosus, Najanaja atra, Sinonatrix annularis, Dinodon septentrionalis, etc. Mater. that the more widespread use of snakes for food and medicine during the Tang It can also be used for skin numbness, dermatosis, or other rashes. By submitting a comment you agree to abide by our Terms and Community Guidelines. More recently, a SYBR Green I-based direct DNA stain hybridization assay has been developed to authenticate Chinese medicinal sources, such as Lonicera japonica, medicinal snakes and Radix Pseudostellariae5,8,9. Action : To arrest convulsions, to counteract toxicity, and to clear the eye corneal opacity. It is common animal medicine, which first appeared in
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