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Background: Assessing the effectiveness and safety of acupuncture for chronic constipation in patients with diabetes mellitus is the main purpose of this systematic review protocol. Methods: The following electronic databases will be searched from their respective inception dates to December 1st 2020: PubMed, the Cochrane Library, Embase, World Science Net, the Allied and Complementary Medicine Database, the Web of Science, China National Knowledge Infrastructure, the Chongqing VIP Chinese Science and Technology Periodical Database and the Wanfang Database. All published randomized controlled trials in English or Chinese related to acupuncture for constipation in patient with diabetes mellitus will be included. The Bristol stool scale, spontaneous complete bowel movements, and observing symptoms (yes/no) including defecation feeling, defecation weakness, feeling of incomplete evacuation, bloating, and flatulence were considered as primary measures. The treatment efficiency consideration according to Bristol stool scale was considered as secondary measure. Two reviewers will conduct the study selection, data extraction and assessment independently. The assessment of risk of bias and data synthesis will be conducted with Review Manager Software (RevMan) V.5.2. Results: The results will provide a high-quality synthesis of current evidence for researchers in this subject area. Conclusion: The conclusion of our study will provide an evidence to judge whether. Acupuncture is an effective intervention for chronic constipation in patients with diabetes mellitus. Ethics and dissemination: Formal ethical approval is not necessary as the data cannot be individualized. The results of this protocol will be disseminated in a peer-reviewed journal or presented at relevant conferences. PROSPERO registration number: INPLASY202110079.

Background Assessing the effectiveness and safety of acupuncture for chronic constipation in patients with diabetes mellitus is the main purpose of this systematic review protocol. Methods The following electronic databases will be searched from their respective inception dates to December 1st 2020: PubMed, the Cochrane Library, Embase, World Science Net, the Allied and Complementary Medicine Database, the Web of Science, China National Knowledge Infrastructure, the Chongqing VIP Chinese Science and Technology Periodical Database and the Wanfang Database. All published randomized controlled trials in English or Chinese related to acupuncture for constipation in patient with diabetes mellitus will be included. The Bristol stool scale, spontaneous complete bowel movements, and observing symptoms (yes/no) including defecation feeling, defecation weakness, feeling of incomplete evacuation, bloating, and flatulence were considered as primary measures. The treatment efficiency consideration according to Bristol stool scale was considered as secondary measure. Two reviewers will conduct the study selection, data extraction and assessment independently. The assessment of risk of bias and data synthesis will be conducted with Review Manager Software (RevMan) V.5.2. Results The results will provide a high-quality synthesis of current evidence for researchers in this subject area. Conclusion The conclusion of our study will provide an evidence to judge whether.Acupuncture is an effective intervention for chronic constipation in patients with diabetes mellitus. Ethics and dissemination Formal ethical approval is not necessary as the data cannot be individualized. The results of this protocol will be disseminated in a peer-reviewed journal or presented at relevant conferences. Prospero registration number INPLASY202110079.

Tongren Dahuoluo Wan has been a popular traditional Chinese medicine in international pharmaceutical markets for hundreds of years. Leopard bone powder is the key element in its formulation. However, the leopard has been listed for wildlife conservation, which limits the use of the leopard bone supplies. Therefore, an alternative formulation which substitutes leopard bone with zokor bone in the formula of Tongren Dahuoluo Wan is now manufactured. To develop a simple and reliable molecular method for authenticating the two patent medicines, mitochondrial nucleotide polymorphic sites of 12S rRNA, COI and Cytb genes were screened in leopard and zokor bones, and nine pairs of species-specific primers were verified for discriminating the two species. For the patent medicine authentication, we set up a molecular diagnostic assay to resolve the difficulties of low concentration of target DNAs and presence of PCR-inhibitory substances in this complex medicine, and successfully confirmed leopard or zokor content using the nine pairs of species-specific primers. We recommend a common technical strategy for authentication of species origins in traditional Chinese medicine, and discuss the experimental solutions for technical problems of molecular diagnostic assays.

Lingyang Qingfei Wan produced by Beijing TongRenTang is a long-standing and popular medicine in China and international pharmaceutical markets. Concerns continue to be raised about the legality of usage of saiga antelope, which was defined as endangered species by Convention on International Trade in Endangered Species of Wild Fauna and Flora legislation and internal legislation in China. Therefore, the alternative pill in which substitutes saiga antelope with goat in the formula of Lingyang Qingfei Wan was developed. In order to authenticate the origin of animal contents in Lingyang Qingfei Wan and its alternative pill, molecular diagnostic assay was utilized by mtDNA polymorphism analysis. Four universal primer pairs containing mtDNA 12SrRNA, 16SrRNA, cytochrome b gene and cytochrome oxidase I were employed to obtain species-specific sequences of saiga antelope and goat, and multiple species-specific primer pairs for saiga antelope and goat were used to identify the animal origin in patent pills according to nucleotide polymorphisms between the two species. In additions, alternative techniques were attempted surrounding dilemmas of low concentration of target DNAs and presence of PCR-inhibitory substances in organic ingredients within complex pill. Results revealed that all species-specific primers could be successfully used for authentication of animal origin within complex pill, and sample preprocessing was critical during experimental manipulation. Internal positive control was an efficient and cost-effective way to assist in monitoring the potential interference from inhibitory substances which existed in the highly processed pills.