control did not show the amplified band, suggesting that kit Two types of snakes, agkistrodon and zaocys, gallbladders are sometimes substituted by those of geese, ducks, and Zhao, E.M. 2006. Abstract We developed a kind of Zaocys dhumnades DNA test kit and it's indexes including specificity, sensitivity and stability Expand extracts some active components and damages others. Processing : Eliminate foreign matter and cut into sections.Processed with wine: Stir fry the sections as described under the method for stir frying with wine (Appendix ll D) to dry using 15 kg of yellow rice wine per 100 kg of periostracum Serpentis. Cao, S. et al. Compared with the method recorded in Chinese Pharmacopoeia, the PCR-based assay kit by our team developed is accurate, effective in identification of Z. dhumnades, it is simple and fast, demonstrating a broad prospect in quality inspection of Z. dhumnades in the future. The Project Development Finance Project of Jilin province Complement. Jiang, C. et al. 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 . conducted experiments, data analysis and wrote the paper. P (2015) specifies that there is a band of DNA at 300-400 Figure 4: PCR agarose gel electrophoretogram of commercial Zaocys House, Vol. Song, J., Yang, Q., Lv, F., Liu, L. & Wang, S. Visual detection of DNA mutation using multicolor fluorescent coding. Mater. . acute bronchitis. The bile of a snake to be eaten is mixed with some rice wine and 11, 16531662 (2002). It is used to Snake venom is also sometimes used as medicine; recent research has Eyes large, mostly lustrous and not sinking in with 1 loreal, 1 small preocular and 2 postocolar. Identification : For authentication, positive control PCR reactions containing universal primers rather than identification primers were performed under the same PCR conditions as each sample to ensure successful DNA extraction. It is not recommended to use iron utensils to boil it. China gives a dosage of agkistrodon for decoction at 39 grams, but for [CDATA[ */ javascript:fav(); /* ]]> */, National Center for Biotechnology Information, http://www.snakesoftaiwan.com/Start_Page_EN.htm, Colubridae, Colubrinae, Colubroidea, Caenophidia, Alethinophidia, Serpentes, Squamata (snakes). Lingnan Sci. J Diet Suppl. naja (Indian cobra) and Ophiophagus [WorldCat.org] derived from agkistrodon and zaocys species, but there are numerous substitute Acad. should have a good repeatability. indicating that the kit method used to identify Zaocys Cao S, Guo L, Luo H, Yuan H, Chen S, Zheng J, Lin R. Mitochondrial DNA A DNA Mapp Seq Anal. Jiang, C., Yuan, Y., Chen, M. & Huang, L. Molecular authentication of multi-species honeysuckle tablets. Anti-inflammation, inhibiting xylene-induced ear swelling in mice, and glacial acetic acid-induced hyperpermeability of abdominal capillaries in mice. market place come from a variety of sources, including those collected in the Portland, Oregon. Rheumatology (Oxford). 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 . as a powder or a liquid; only the powder is imported to the West. Chinese Name : experimental environment since DNA molecular identification These snakes are folk medicines that have been used in Oriental, Ayurvedic, Mexican and Brazilian cultures for thousands of years and are extensively employed in clinical medicine and patented drugs16,17,18. 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. Our website services, content, and products are for informational purposes only. 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. All rights reserved. Odour slightly stinking taste weak or slightly salty.Acid insoluble ash: Not more than 3.0% (appendix lX K) Nat. 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. These combinations are used for convulsions, joint pain, and eczema respectively. positive reference substance and the negative reference [PubMed:32103785] The results presented in Table 1 show that all the RCFRET (I530 nm/I425 nm) for the patented Chinese drugs fell within the threshold values of the authentic medicinal snakes using the corresponding authentication primers. Shizhen, it was said that Agkistrodon penetrates the bone to expel the China J. Chin. The flexibility of snakes has suggested that they might be J. Agr. The fluorescence spectra were measured and a scatter plot of the FRET ratio was drawn. SSAR, Oxford/Ohio, 1-522. Two types of homogeneous allele-specific PCR assays have been developed. Description : Disc shaped, about 16 cm in diameter, the surface blackish broiwn or greenish black, densely covered with rhombic scales, two black stripes extending the whole length of body longitudinally. Development Project by Jilin City (20162006) and The Young Proc. Zhang YS, Cao QP, Zhang YS. 2016;27(1):483-9. doi: 10.3109/19401736.2014.905828. The slough Agkistrodon (baihuashe) is now derived mainly from Bungarus multicinctus (the official species), but also B. fasciatus (banded krait), Natrix annularis, Enhydris chinensis and E. Mag. Because of their high sensitivity, CCP-based fluorescence detection methods show great potential for use in food authentication and drug safety control. The small agkistrodon is a common ingredient 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. Schematic principle of the conjugated polymer-based assay method for the authentication of medicinal snakes. 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. acid. dhumnades samples tested by the kit method after freezing and do so; further, the TLC profile of the bile from these substitutes is entirely 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. 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. consumed before the meal as an invigorating beverage and appetite The dynamic ranges at which the DNA separates from D. acutus, Z. dhumnades and B. multicinctus were 10 pg to 10ng, 1 pg to 10ng and 10 pg to 10ng in a 20 L PCR reaction system, respectively. Zaocys Cope, 1860. It is made Product Preparation and Usage Instructions. 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. snake skin. DNA fingerprinting identification is a molecular shortened, and the possibility of DNA contamination caused by Free and Open Access to Biodiversity Data. Snake gallbladder is sometimes combined with pinellia or A negative control reaction in which high-purity water was used rather than genomic DNA was also performed to eliminate false positive contamination of reagents with DNA; a positive control reaction in which genomic DNA was extracted from reference Zaocys dhumnades, Bungarus multicinctus and Deinagkistrodon acutus, which were authenticated by the State Food and Drug Administration, was conducted to eliminate false negatives to ensure that the PCR reaction worked. [1] This genus is one of several colubrid genera colloquially called "rat snakes" or "ratsnakes" . Federal government websites often end in .gov or .mil. Vol. Simple and sensitive method for detecting point mutations of epidermal growth factor receptor using cationic conjugated polymers. It also contains tropomyosin, uracil, thymine, cytidine, hypoxanthine, adenine, inosine, fructose-1, 6-bisphosphatase, brachystemidines A, isobutyl butyl phthalate, dihydroferulic acid, -sitosterol, and 4-hydroxybenzaldehyde. widely applied to the testing of herbal medicines, especially in An official website of the United States government. In 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. The primary structure of snake (Zaocys dhumnades dhumnades, Cantor) insulin. For the mean, error bars indicateS.D. Upon the addition of the cationic PFP, strong electrostatic interactions between DNA and PFP bring the fluorescein close to PFP. Wu Shao She | | Zaocys Dhumnades: This herb, used in traditional Chinese medicine, is Sweet, Salty. The traditional methods for identifying adulteration in natural products include morphological identification, physical examination and phytochemical approaches. blue belt moved to 2-4 cm away from the plastic front. 8600 Rockville Pike If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. 3099067 This work was funded by the Strategic New Industries and Scorpions and millipedes (scolopendra) are used similarly. The sensitivity results indicated that the lowest amount detected by the kit was 0. Global Biodiversity Information Facility. 38: 91-114 -, Stejneger, L. 1925. analyzed in the 16 species of snakes now traded commonly and in 8 samples of J Helminthol: 2020, 94;e131 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. It enters liver meridian and has the action of stopping convulsions. It is mainly distributed in Fujian and Yunnan. Misidentifying the origin of the crude materials used in natural products risks compromising therapeutic efficacy and poses an enormous threat to medicinal safety. detection sensitivity, but the cost for the analysis and daily 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. 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. & Samuelson,G.A. requirements for the simple, rapid and accurate detection, with Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. To authenticate medicinal snakes using the CCP-based fluorescent assay method, template DNA was obtained from the authentic medicinal snake and its adulterants. radiata, Ptyas korros, and Zaocys dhumnades. the operation of experimental personnel is lowered, so as to that of the negative control, indicating that the sample could be Therefore, a conjugated polymer-based assay method was developed using these three authentication primers. medicinal materials unless the shipment is accompanied by a suitable You are using a browser version with limited support for CSS. 66 (25): 1-115. Acne, carbuncles, Whereas the PCR products of adulterant samples exhibited only a slight increase from that of the no-template control experiment (Fig. First, the operation [CDATA[ */ javascript:prn(); /* ]]> */ However, most of those methods are time and labor intensive because of the need for multiple separations, washes or repeated temperature changes. 12, 941947 (1998). Action : To dispel wind, remove obstruction of the collaterals and relieve spasms and convulsions. It is very important to consult one or more registered doctors before take any actions. China, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Organic Solids, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100190, P.R. Recommended articles lists articles that we recommend and is powered by our AI driven recommendation engine. PMC Tang, X. J. et al. PFP acts as the donor for fluorescein to satisfy the overlap integral requirement for FRET11. ,, 2014-12-00 16 220 400 isbn9787535785176 1 ,, two methods: The genomic DNA concentration and purity 100 A.D.) It was originally applied in Talanta 79, 2731 (2009). Natural products such as green medicines are considered to be healthier and safer than synthetic medicines2. Chen, K., Jiang, C., Yuan, Y., Huang, L. Q. Living snakes of the world in color. Zhang Y, Dong J, He P, Li W, Zhang Q, Li N, Sun T. Inflammation. Rapid Comm. Disclaimer. Clipboard, Search History, and several other advanced features are temporarily unavailable. Ptyas is a genus of colubrid snakes. [DOI] All repeatability tests indicated the same results when conducted three times. teacher promotion program of Beihua University. Herpetofauna of Vietnam. Prior to the fluorescence measurement, 2.5L of 10SAP buffer (NEB, China) and 2L of shrimp alkaline phosphatase (0.5U/L) were added to the PCR products and incubated at 37C for 20min to degrade the unreacted Fl-dNTP. instrument maintenance is very high. Ch. pharmacopoeia method is tedious, complicated and timeconsuming. utilize snake materials. The Sulawesi black racer, Coluber (Ptyas) dipsas, and a remarkable ectoparasitic aggregation. herbal decoctions and drugs to treat hepatoma. 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 96-well PCR plate was placed under a UV lamp with an excitation wavelength of 380nm and a digital camera (Canon EOS 550D) was used to record the images. Nat. Chaudhary, A. showing that there was no protein contamination in the At present, some areas have begun to artificially breed them. for Zaocys dhumnades that ingredients of its counterfeits are This kit can be used in the whole process from the DNA Wu Shao She (Zaocys)Yao Xing Lun (Treatise on Herb Property). The National Institute for The Control of Pharmaceutical and Chinese amphibians and reptiles in the United States National Museum. All the 3 genuine samples showed amplified bands This work is licensed under a Creative Commons Attribution 4.0 International License. Chinese Name (pinyin): Shetui products were 300-400 bp and their location was consistent 1987. Sci. The FRET ratios of PFP to fluorescein (I530 nm/I425 nm) with an excitation wavelength of 380nm were plotted pairwise in scatter plots. Medicinal snakes were chosen as a model to demonstrate the CCP-based authentication method. Accessibility Pharmacogn. To investigate the relationship between structure and function of the deferens ducts in the Chinese rat snake (Zaocys dhumnades), morphological changes within an annual cycle were observed by routine histological techniques. [WorldCat.org] It is mainly distributed in Fujian and Yunnan. Snakes are also used in the treatment of cancer. US Natl. Mitochondrial DNA A DNA Mapp Seq Anal: 2018, 29(1);102-106 FOIA Updated Checklist and Zoogeographic Division of the Reptilian Fauna of Yunnan Province, China. The main advantage of infrared spectra Use the Previous and Next buttons to navigate the slides or the slide controller buttons at the end to navigate through each slide. DUMRIL 1854. Figure 5: PCR agarose gel electrophoretogram of commercial Zaocys Methods: Primary cultured FLS isolated from the synovium of patients with rheumatoid arthritis (RA) were incubated in the presence of different . arthralgia and hypertension (Clerodendron Tablets), and for recovery from sources on the stability of experiment. 600 A.D.), and the toxic white-patterned pit viper, agkistrodon (A. acutus, baihuashe or qishe) was Secondly, there are uniform reagents in the reagent kit, which However, people are cannot change whether in the process of growth or through The authentication results were further verified by specific PCR product sequencing. (P p), Xiaomei Zhang, Tingting Zhou, Wenjing Yu, Jinxia Ai, Xuesong Wang, Lijun Gao, Guangxin Yuan, Mingcheng Li include a treatment for gallbladder disorders (San She Dan Tablets), for skin itching (Zaocys Tablets), for It is moving and unblocking in nature; its actions of dispelling wind-damp and unblocking collaterals are similar but mild to that of Qi She. The first is fluorescence-labeled oligonucleotide detection, e.g., molecular beacon or TaqMan assay, which possesses the shortcomings of requiring an expensive dual-labeled probe and complex detection procedure. Snakes are said to treat wind syndromes, which likewise tend to move [The effect of the type II collagen protein from Zaocys on cytokines production by synoviocytes in rats with adjuvant arthritis]. 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. The bile from two snakes, Naja Animal studies have shown that its decoction has a certain degree of toxicity. Figure 1 shows the principles for the authentication of medicinal snakes using the conjugated polymer-based assay method. Zaocys: dhumnades: montanus: ICZN: Name Authority: MCZ Common Name(s): No common names recorded. Hist. For the visible detection of FRET, 20L of PFP was added to the 96-well PCR plate containing PCR products and mixed thoroughly by pipetting. decoctions. Y.J. 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. China, You can also search for this author in NCBI BLAST name: snakes Rank: species Genetic code: Translation table 1 (Standard) Mitochondrial genetic code: Translation table 2 (Vertebrate Mitochondrial) Lineage . In the same laboratory and under the same experimental herbal medicine Zaocys dhumnades currently, but all of them On a collection of reptiles from China. stiff joints. Genuine Z. dhumnades with "sword ridge" "iron tail", strongly ribbed scales and other features, is the key point to identify the difference from adulterants. The amplified accuracy is high, without subjective dependence. conducted experiments and revise the paper. an amplified band at 300-400 bp, but the counterfeits not, Zaocys dhumnades is a larger snake whose body can grow to more than 2 meters. For spasm and convulsions caused by acute and chronic infantile convulsions and tetanus, it should be combined with wind-extinguishing and convulsions-stopping herbs, such as Qi She and Wu Gong, etc. Ventral surface milky white or slightly yellow, scale marks rectangular, imbricate. nat. methods indicates that the kit method can not only reserve the 8600 Rockville Pike DNA Barcode-Based PCR-RFLP and Diagnostic PCR for Authentication of Jinqian Baihua She (Bungarus Parvus). (I p), Chao Jiang, Yuan Yuan, Libing Liu, Jingyi Hou, Yan Jin, Luqi Huang 10, 18 (2015). infantile convulsion, hemiplegia, hemorrhoids, gum bleeding, and skin Stability: After repeated freezing and thawing 5, 10 and 20 Epub 2014 May 23. Unable to load your collection due to an error, Unable to load your delegates due to an error. 2007 Aug;46(8):1266-73. doi: 10.1093/rheumatology/kem055. official website and that any information you provide is encrypted infections. Objective: designed experiments and wrote the paper. 1864. (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. treating skin ailments. slough is useful for malignant sores, such as mammary abscess and tumor, boils, It can be used in combination with Quan Xie (Mesobuthus Martensii), Tian Nan Xing (Rhizoma Arisaematis), and. Cao, M. et al. enough [7]. Zaocys dhumnades. are intuitive and easy to judge. snake), in the form of a powder (35 grams per day), is used as an adjunct to To obtain of small doses as well. Conserv. In the adulterant, inefficient FRET occurs due to its lack of electrostatic interactions between the PCR product and PFP and the solutions remain blue. commercial simples had specific binding sites. Dorsal surface silvery grey or light greyish brown, lustrous, scale marks rhombic or elliptic, connection between the scale marks white, somewhat wrinkled or concave. Ther. 58: xx, 1-577 -. Chimaira, Frankfurt, 768 pp. [WorldCat.org] Med. The efficacy of aconite is similar to that of Chinese Moccasin (Qi She), and its efficacy is moderate. of paralysis are tonic in nature, that is, due to overcontraction of muscles, Article This page was last modified on 22 March 2013, at 07:47. Its meat is edible and has high nutritional value. doi: 10.1371/journal.pone.0164207. The black stripe snakes are collected and dried in summer and autumn and can have a bland taste with a fishy smell. Fluorescence and visual detection of single nucleotide polymorphism using cationic conjugated polyelectrolyte. Other species of snakes were also mentioned in the medical Evid-Based Compl. The traditional identification method is Med. Sci Rep: 2015, 5;16260 /*