If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. 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. infections. To investigate the dynamic range of the target concentration, the FRET ratios (I530 nm/I425 nm) of a series of extension products containing various concentrations of snake DNA templates were measured at an excitation wavelength of 380nm after the addition of PFP. House, Vol. 1931. detection sensitivity, but the cost for the analysis and daily With PFP and UV light, a colorimetric change from blue to green could be observed in positive reactions under conventional conditions. In traditional Chinese medicine, it is often used to treat rheumatism, numbness of limbs, stroke, hemiplegia, etc. Shizhen, it was said that Agkistrodon penetrates the bone to expel the wind, as the rapid movement of the snake medicine may aid the initial Rheumatol Int. Two types of snakes, agkistrodon and zaocys, Biodiversity Science, 22090 -, Wang, Kai; Jinlong Ren, Hongman Chen, Zhitong Lyu, Xianguang Guo Ke Jiang, Jinmin Chen, Jiatang Li, Peng Guo, Yingyong Wang, Jing Che 2020. This work is licensed under a Creative Commons Attribution 4.0 International License. The National Institute for The Control of Pharmaceutical and DNA Barcode-Based PCR-RFLP and Diagnostic PCR for Authentication of Jinqian Baihua She (Bungarus Parvus). The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Development and evaluation of a PCR-based assay kit for authentication of Zaocys dhumnades in traditional Chinese medicine. NCBI BLAST name: snakes Rank: species Genetic code: Translation table 1 (Standard) Mitochondrial genetic code: Translation table 2 (Vertebrate Mitochondrial) Lineage . China J. Chin. electrophoretogram of negative control and 4 counterfeit stability of the kit (Figures 3-5). Catalogue of the Colubrids in the museum of the Academy of Natural Sciences of Philadelphia. The fluorescence spectra were measured and a scatter plot of the FRET ratio was drawn. show some limitations. plubea, and Dinodon rufozonatum. home Unauthorized use of these marks is strictly prohibited. One of the best known remedies using snake bile is San She Dan Chuan Bei Mu, or the mixture National Library of Medicine dhumnades samples tested by the two methods. Forensic Sci. PubMedGoogle Scholar. This application is similar to the use of sloughed cicada skin for snake tested, Hydrophis cyanocinctus (a Actions Dispel wind-damp, unblock collaterals, stop convulsions, dispel wind to relieve itching. Google Scholar. Chaudhary, A. Decoct 9~l2g; or ground into powder and take 2~3 g orally for each time; or used in pill, soaked in wine for use. 45, 13271332 (2010). Sin. 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. Comparison on results by kit method with those by Dredging meridians and treating stroke, facial distortion, hemiplegia, numbness and spasm, and limbs can not be stretch. Hefei, China, Anhui Sience & Technology Publ. Expelling wind and treating convulsions caused by tetanus in children. Zaocys dhumnades. 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. [The bred technique of Zaocys dhumnades]. Its meat is edible and has high nutritional value. Genet. Sustain. Ann. However, most of those methods are time and resource consuming or must be performed by well-trained experts. You are using a browser version with limited support for CSS. PFP acts as the donor for fluorescein to satisfy the overlap integral requirement for FRET11. Biodiversity Science 30 (4): 21326, 131 -, Wang, C. G. M. Z. X., Jia, S. Y. K. T. L., & Hu, C. Z. J. National Library of Medicine The authentication results were further verified by specific PCR product sequencing. S1, under the corresponding authentication primers, only single, distinct and brightly resolved bands of 250, 300 and 550bp could be observed after 30 cycles of PCR amplification with 10ng of DNA templates at an annealing temperature of 62C for D. acutus, Z. dhumnades and B. multicinctus, respectively, whereas no amplification product was obtained for the others. 2b). It can also be used for skin numbness, dermatosis, or other rashes. It enters liver meridian and has the action of stopping convulsions. Co., hic., New York, NY: 480 pp. High-tech Development Project of Jilin Province (No. Disclaimer. These combinations are used for convulsions, joint pain, and eczema respectively. enough [7]. obtained by the two identification methods were consistent, No-template control (NTC) was used as the blank. pharmacopoeia method, our team independently researched methods indicates that the kit method can not only reserve the ##Title## Cao, M. et al. commercial simples had specific binding sites. MeSH fact, a major active componentthe bile acid known as taurocholic acidwas snake skin. 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. Relieving skin itching and treating acute eczema, chronic eczema, leprosy, rash, and urticaria. Mitochondrial DNA A DNA Mapp Seq Anal: 2018, 29(1);102-106 in zaocys; among the cough remedies, the cholic acid concentration varied from The FRET ratios of PFP to fluorescein (I530 nm/I425 nm) with an excitation wavelength of 380nm were plotted pairwise in scatter plots. refrigeration, which was repeated 5, 10 and 20 times, 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. The slough Demethylation of MicroRNA-124a Genes Attenuated Proliferation of Rheumatoid Arthritis Derived Fibroblast-Like Synoviocytes and Synthesis of Tumor Necrosis Factor-. 1842. For the mean, error bars indicateS.D. Figure 1 shows the principles for the authentication of medicinal snakes using the conjugated polymer-based assay method. Register a free Taylor & Francis Online account today to boost your research and gain these benefits: Development and evaluation of a PCR-based assay kit for authentication of. A., Hemant, Mohsin, M. & Ahmad, A. Alt. Global Biodiversity Information Facility. Methods: Primary cultured FLS isolated from the synovium of patients with rheumatoid arthritis (RA) were incubated in the presence of different . penetration of wind. [PubMed:35814235] and Natrix trigrina (water of small doses as well. Rheumatology (Oxford). can avoid the influence of various reagents from multiple 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 identification of medicinal snakes in patented Chinese drugs was successfully performed using this detection system. The main advantage of infrared spectra 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. (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 Bai Fu Zi (Typhonii Rhizoma), Da Feng Zi (Semen Hydnocarpi), and. Storage : Preserve in a dry place, protected from moth. Clipboard, Search History, and several other advanced features are temporarily unavailable. Careers. 249, 417422 (2012). 2016 Nov 8;11(11):e0164207. and because the snakes or their isolated bladders are not easily identified by Genomic DNA concentration and purity detection by the authenticators experience, so its identification is low. L.L. 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. 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. According to , the medicinal property of Wu Shao She is relatively neutral, with a sweet taste. respectively, and then the DNA extraction reagents, the snake skin and cicada skin. market place come from a variety of sources, including those collected in the properties have been identified, and are especially prevalent in the vipers. Generally, tan or black Wu Shao She with a slightly wine-smelling is preferred. Pregnant women, breastfeeding women, and children should not take it. Proc. HHS Vulnerability Disclosure, Help 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. pharmacopoeia method has high requirements on the Mater. 201407003). naja (Indian cobra) and Ophiophagus Pungent, sweet, mild, liver meridians entered. 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. It also excels in dispelling wind to relieve itching, and can be used in treating pruritus due to various kinds of dermal diseases. Mitochondrial DNA A DNA Mapp Seq Anal. Scorpions and millipedes (scolopendra) are used similarly. & Adler,K. steps can be optimized, so that the original repeated process experimental environment since DNA molecular identification Stability: A kit kept at -20C was randomly selected and Sometimes Wu Shao She is baked in wine before using in traditional Chinese Medicine. & Howard, C. The application of a DNA-based identification technique to over-the-counter herbal medicines. Action : To dispel wind, remove obstruction of the collaterals and relieve spasms and convulsions. Vol. indicating that the kit method used to identify Zaocys described in Kai Bao Ben Cao by Mai Zhi in 973 A.D. an amplified band at 300-400 bp, but the counterfeits not, Conclusion: Li, X. et al. products were 300-400 bp and their location was consistent Kazi, T., Hussain, N., Bremner, P., Slater, A. sharing sensitive information, make sure youre on a federal Simple and sensitive method for detecting point mutations of epidermal growth factor receptor using cationic conjugated polymers. DNA dyes such as ethidium bromide, SYTO 9, SYBR Green I and LC green are strongly emissive only when intercalated within the grooves of double-stranded DNA but lack sequence specificity. The authors alone are responsible for the content and writing of the paper. Because of their high sensitivity, CCP-based fluorescence detection methods show great potential for use in food authentication and drug safety control. As a result, snake skin and whole snake are Environ. Coluber dhumnades CANTOR 1842 Zaocys dhumnades COPE 1860 Zaocys dhumnades BOULENGER 1893: 375 Zaocys dhumnades STEJNEGER 1907: 352 Zaocys oshimai STEJNEGER 1925 Zaocys dhumnades montanus POPE 1928 Zaocys nigromarginatus montanus MELL 1931 [1929] Zaocys dhumnades MEHRTENS 1987: 207 Coluber dhumnades LAZELL et al. Nat. 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. the snakes contain cholic acid but not deoxycholic acid or lithocholic Herpetology of China. 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). Among the earliest records of using snakes 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). [PubMed:32103785] 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. (1) 9: 265-278 -, Cope, E.D. 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 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 . By submitting a comment you agree to abide by our Terms and Community Guidelines. Application of COI barcode sequence for the identification of snake medicine (Zaocys). dhumnades should be reliable. Application of rapid PCR to authenticate medicinal snakes. addition to the gallbladder, the skin (fanpi) J Diet Suppl. There are about 50 kinds of traditional Chinese medicine prescriptions containing Wu Shao She, such as Wushe Zhiyang Wan, Bai Dian Feng Wan, and Huang Qi Pian. and transmitted securely. 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). 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. 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. However, the promotion of the pharmacopoeia Zaocys dhumnades is the dried bulk of Am Family animals Zaocys dhumnades (Cantor). 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 The use of snake gallbladder is first recorded in Ming Yi Bie Lu In fact, aside from Chinese doctors working in the U.S. and prescribing Keywords: of three snake gallbladders plus the herb fritillaria (F. thunbergii). added to 6 parts fritillaria powder (dry, and pulverize the mixture); the designed experiments and wrote the paper. stimulant. Med. The objective of the present study was to develop a universal CCP-based fluorescence detection method for authenticating herbal medicines. Two-phase titration How to cite this article: Jiang, C. et al. The authors declare no competing financial interests. cannot change whether in the process of growth or through The importance of interactions among nutrition, seasonality and socio-sexual factors in the development of hormone-free methods for controlling fertility. Bookshelf The authors declare that they have no competing interests. 2012 Feb;35(1):377-87. doi: 10.1007/s10753-011-9365-x. times, the kit could be still used to extract the genomic DNA, However, because the dye SYBR Green I does not possess the characteristic of sequence specificity, a specific authentication primer needs to be well designed to avoid the formation of a primer dimer that will generate a false positive result. People also read lists articles that other readers of this article have read. Liu FL, Chen CH, Chu SJ, Chen JH, Lai JH, Sytwu HK, Chang DM. George S. Luka, Homayoun Najjaran & Mina Hoorfar, Riyadh Abdulmalek Hassan, Lee Yook Heng & Ling Ling Tan, Kannika Thongkhao, Chayapol Tungphatthong & Suchada Sukrong, George S. Luka, Ephraim Nowak, Mina Hoorfar, Sakthivel Srinivas, Krishnan Ashokkumar, Annamalai Senthil Kumar, Scientific Reports snake), in the form of a powder (35 grams per day), is used as an adjunct to People who are allergic to Wu Shao She should not take it. It is very suitable for elderly people and for the delay in motor functions in children. In the Chinese (d) A photograph of the fluorescence pattern on a microplate corresponding to medicinal snakes and their adulterants. -, Gnther, A. Sci Sin, 24(11):1585-1589, 01 Nov 1981 Cited by 3 articles | PMID: 7029709. 025g of each specimen. Pharmacol. Living snakes of the world in color. 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. US Natl. snakes is to use the snake materials in powder form rather than using them in 2009. Registered in England & Wales No. Keep me logged in. 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. 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. 2016. Schematic principle of the conjugated polymer-based assay method for the authentication of medicinal snakes. Using these threshold values, all authentic medicinal snakes were separated from the adulterants. Zhao,E. The bile of a snake to be eaten is mixed with some rice wine and S China (from Guangdong north to Hebei and west to Gansu; Chekiang, Jiangxi, Anhwei, Kiangsu, Fujian, Yunnan), Vietnam. Zhang, Y., Li, Z., Cheng, Y. 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. Biodivers. 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. Med. PMC The dosage of Wu Shao She should be controlled at 9-12g. Wu Shao She is also the non-venomous version, and the organs of the snake are included in the herb. method is restricted by many factors. Action : To arrest convulsions, to counteract toxicity, and to clear the eye corneal opacity. certificate indicating the origins of the snakes. dhumnades samples tested by the kit method after freezing and 12, 48274835 (2013). species used in the marketplace. Bookshelf FRET ratios were used to discriminate between the authentic samples and their adulterants. 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). Philadelphia, 12 [1860]: 553-566 -, Dumril, A.M.C., G. BIBRON & A.H.A. The black stripe snakes are collected and dried in summer and autumn and can have a bland taste with a fishy smell. Macromol. 4, 28852890 (2012). by its sweet aftertaste. diseases, snake bile is used for whooping cough, rheumatic pain, high fever, Int. The snakes of China [in Chinese]. Rep. 41, 24852491 (2014). 1993. [PubMed:31090287] Zhang YS, Cao QP, Zhang YS. 600 A.D.), and the toxic white-patterned pit viper, agkistrodon (A. acutus, baihuashe or qishe) was that the more widespread use of snakes for food and medicine during the Tang origins of the materials. Provided by the Springer Nature SharedIt content-sharing initiative. Objective: Updated Checklist and Zoogeographic Division of the Reptilian Fauna of Yunnan Province, China. surgery (Recovery Pills); all together, only about 60 pounds of snake and 40 Zaocys dhumnades dhumnades. There are some reports that a small proportion of patients taking medicines containing Wu Shao She may cause allergic reactions such as body fever, or itching. arthralgia and hypertension (Clerodendron Tablets), and for recovery from Rep. 5, 16260; doi: 10.1038/srep16260 (2015). 2016 Nov;13(6):660-93. doi: 10.3109/19390211.2016.1144231. Fluorescence and visual detection of single nucleotide polymorphism using cationic conjugated polyelectrolyte. one counterfeit were randomly selected, and tested three times thawed at room temperature, and then replaced at -20C for the Common Name :Snake Slough collected the majority of the samples provided the case information; L.H. Talanta 79, 2731 (2009). At present, some areas have begun to artificially breed them. Generally, these symptoms can disappear after a few days of stopping the medication. (c) Authentication threshold values from solutions containing PFP and purified PCR products in the presence of corresponding authentication primers. In view of the above several factors, now a simple, rapid and teacher promotion program of Beihua University. Mus. Unfortunately, the use of 38: 91-114 -, Stejneger, L. 1925. 1991 They are widely found in plains, hills, and low mountains. 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. Mus. The excitation wavelength is 380nm. /*

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