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Objective::To establish a pre-column derivatization reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of 17 amino acids in Cynomorii Herba from different producing areas and conduct a multivariate statistical analysis. Method::RP-HPLC with pre-column derivatization was employed, with phenyl isothiocyanate (PITC) as derivatization reagent. Separation was performed on a WondaSil C18-WR column (4.6 mm×150 mm, 5 μm), with 0.05 mol·L-1 sodium acetate solution (pH 6.5) as mobile phase A, and acetonitrile-methanol-water (3∶1∶1) as mobile phase B for gradient elution at a flow rate of 0.8 mL·min-1. The detective wave length was set at 254 nm, and the column temperature was maintained at 35 ℃. Principal component analysis (PCA) and systematic cluster analysis (HCA) models were established for multivariate statistical analysis and quality evaluation. Result::17 Kinds of amino acid were detected in Cynomorii Herba, 7 of which were essential amino acids. The 17 amino acids showed good linearity in respective concentration range, r = 0.999 0-0.999 9.The average recoveries were between 98.03%-103.89%with RSD<3.5%. The results of PCA and HCA were basically the same, and both methods can be used to clearly distinguish Cynomorii Herba from 12 municipal producing areas into 6 regions. PCA can be used to classify Cynomorii Herba according to different municipal or provincial production areas, and HCA can be used to classify it according to provincial production areas. It showed that the amino acid contents in Cynomorii Herba from different municipal and provincial producing areas had differences, and the content distribution showed obvious geographical clustering characteristics. PCA showed that Cynomorii Herba from Gansu province and Inner Mongolia had higher amino acid contents and better quality as compared with other producing areas. Conclusion::The established method can be used for content determination of 17 amino acids in Cynomorii Herba from different producing areas, and provide a reference for its comprehensive quality evaluation.
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OBJECTIVE@#To study the effects of Angelicaesinensis radix (Danggui) decoction on the therapeutic action and the colonic morphology and mucus secretion in XuexuBianmi model mice.@*METHODS@#Sixty Kunming mice, maleandfemaleinhalf, were randomly divided into six groups according to gender and weight (=10):normal control group, XuexuBianmi model group, positive control group, and high-dose, middle-dose and low-dose Danggui groups. Except the normal control group, the mice in the rest groups were orally administrated with diphenoxylate (DPN) and subcutaneously injected with acetylphenyhydrazine (APH) and intraperitoneally injected with cyclophosphamide (CPA) to copy XuexuBianmi model. Fromthe 14th day, the mice in Danggui groups were orally administered with different doses of Danggui decoction (16.67、8.33、4.17 g/kg), the mice in positive control group were orally administered with Changtongshu granule (5 g/kg), the mice in XuexuBianmi model group were administered with normal saline (NS) at the same volume (10 ml/kg), once a day for consecutive 28 days. The general status were observed, the first black defecation time (FBDT), the water content in stool and in colon in mice of all groups were tested. And the colon tissue was stained with hematoxylin-eosin (HE) and AB-PAS to observe the changes of colonic morphology and the mucus secretion.@*RESULTS@#Compared with the normal control group, there appeared the XuexuBianmi syndromes in model group as follows, the defecation time (FBDT) was significantly prolonged, the water contentinstool and in colon were decreased (<0.01), the colonic mucosa and gland were atrophied, mucous membrane layer was thinned (<0.01), mucus secretion was decreased. Compared with the XuexuBianmimodel group, the Xuexu and Bianmi syndromes were improved, the FBDT was significantly shortened (<0.05, <0.01), the water content of colon was increased in the three groups of Danggui decoction (<0.05, <0.01). The water content of the stool was obviouslyincreased in groups of 16.67and 8.33kg.dDanggui decoction (<0.05). The atrophy of colon mucosa and gland was improved, the mucus secretion was increased, and the colon lubrication function was improved in the three groups in different doses of Danggui decoction.@*CONCLUSIONS@#Dangguican improve the mucosal atrophy, and increase the secretion of colonic mucus, then the stool is softened and promoted to discharge.
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Animals , Mice , Angelica sinensis , Colon , Drugs, Chinese Herbal , Mucus , Plant RootsABSTRACT
Objective To optimize prescription of gentiopicroside nanoemulsion; To evaluate its quality and security. Methods According to the basic properties of gentiopicroside, single-factor test and pseudo-ternary phase diagram were adopted to optimize prescription of gentiopicroside nanoemulsion. The physicochemical property was investigated, and the content was evaluated by HPLC. Results Optimal prescription was obtained as gentiopicroside/ethyl acetate/ EL-35/ absolute ethyl alcohol/ ultrapure water (1.77:7.66:5.94:11.91:72.71); the particle size of nanoemulsion was 23.08 nm; the content of gentiopicroside was determined by HPLC; the specificity of the method was good with a linear range of 0.004 28–0.214 mg/mL; stability of prepared gentiopicroside nanoemulsion was good in the state of different temperatures, illumination intensity and humidity conditions, without significant changes. Conclusion The optimized preparation technology of gentiopicroside nanoemulsion is simple, repeatable and stable, which can provide references for improving the bioavailability of gentiopicroside.
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Objective To observe the effects of Angelica Sinensis Radix on the expression of aquaporin 8 (AQP8) and the levels of AC-cAMP-PKA in colon of blood-deficiency constipation in mice; To identify mechanism of Angelica Sinensis Radix for loosening the bowels to relieve constipation. Methods Sixty Kunming mice were randomly divided into control group, model group, positive medicine group, and Angelica Sinensis Radix high-, medium-, and low-dose groups, with ten mice in each group. Diphenoxylate, acetylphenlyhydrazine and cyclophosphamide were used to establish blood-deficiency constipation mice models. From the 14thday of the experiment, Angelica Sinensis Radix high-, medium-, and low-dose groups were given 16.7, 8.8, and 4.2 g/kg Angelica Sinensis Radix Decoction for gavage. Positive medicine group was given 5.0 g/kg Changtongshu Granules Liquid for gavage. Control group and model group were given equal volume of saline for gavage. The symptoms of blood deficiency constipation were observed and defecation time. Immunohistochemistry, Western blot and qRT-PCR were used to detect the expression of AQP8 protein and mRNA in colon, and the expression of AC-cAMP-PKA in colon was detected by ELISA. Results The mice in the model group developed blood deficiency constipation syndrome; the defecation time was significantly prolonged (P＜0.01); the expression level of colonic AQP8 protein and mRNA, AC, cAMP and PKA significantly increased (P＜0.05, P＜0.01). Compared with model group, the defecation time was significantly shortened in Angelica Sinensis Radix high-, medium-, and low-dose groups; the expression of AQP8 protein and mRNA and the levels of AC, cAMP and PKA were significantly decreased (P＜0.05, P＜0.01). Conclusion The treatment of Angelica Sinensis Radix for blood-deficiency constipation may be related to adjusting the AC-cAMP-PKA signaling pathways and reducing the expression of AQP8 protein and mRNA.
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BACKGROUND: There are a variety of methods for establishing periprosthetic osteolysis animal model, and the models established by different methods and with different animals have their own characteristics.OBJECTIVE: To review the research profiles of the periprosthetic osteolysis and the characteristics of the various models, and to provide a reference for the related research. METHODS: The relevant articles on the periprosthetic osteolysis animal models were searched in PubMed from January 1999 to May 2017, with the key words of "animal model, osteolysis" in English by the computer. Similarly, Chinese Journal Full-text Database was retrieved for related articles published from January 1999 to May 2017, with the key words of "animal model, osteolysis" in Chinese. Articles that were unrelated to the periprosthetic osteolysis animal model were excluded. Classic and recently published articles were selected. RESULTS AND CONCLUSION: (1) The artificial induced animal models are the principal means to establish animal models of periprosthetic osteolysis and they have been widely used in the study of periprosthetic osteolysis pathogenesis, pathological process and treatment. (2) Small-animal models present many advantages: inexpensive to purchase, high physiological turnover, controlled source, homogenous and established genetic background. However, a limitation of these small-animal models is that the temporal pattern of osteolysis is different from the long-term osteolysis that occurs in patients. (3) The large animal model is more similar to the bone dissolution process around the human prosthesis, but the cost is high; the breeding is difficult; the operation requirement is hard. Thus, its widespread use is limited. (4) Furthermore, innovative models that combine the advantages of both are also constantly explored. Overall, the selection of models should be based on research conditions and needs to be considered.
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The methods of literature metrology and data mining were used to study the research topics and social network analysis of traditional Chinese medicine for constipation. The major Chinese databases were searched to include the research studies of traditional Chinese medicine for constipation. BICOMS analysis software was used to extract and collect the main information and produce co-occurrence Matrix; gCLUTO software was used for cluster analysis. Data analysis was conducted by using SPSS 19.0 software. The results showed that the number of studies on traditional Chinese medicine for constipation was constantly increased, with two literature volume peaks respectively in 2003 and 2006. Related studies have been published in 31 provinces, autonomous regions and municipalities have published, but the studies in developed areas were more than those in developing areas. There was little cooperation between research institutions and the authors, especially the cooperation between different areas. At present, the research field of Chinese medicine for constipation is divided into five research topics. In terms of specific traditional Chinese medicine, angelica sinensis is in the core position. The results showed regional imbalance in the number of studies on Chinese medicine treatment for constipation, as well as little cooperation between researchers and research institutions. The research topics mainly focused on the evaluation of clinical efficacy, but the research on optimizing the prescriptions was still not enough, so the future researchers shall pay more attention to the studies of constipation prescriptions with Angelica sinensis as the core herb.
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Objective: To optimize the preparation methods of diabetic gastroparesis rat model by orthogonal test. Methods: The diabetes gastroparesis rat model was duplicated with alloxan (ALX) by using four factors and three levels of orthogonal design. The fasting blood glucose, gastric emptying rate, and plasma motilin level were scored as evaluation indexes, and the influence of different factors on diabetic gastroparesis model rats was observed. Results: The effect of feed type on the diabetic gastroparesis rat model was not obvious, but the effects of modeling time and modeling agent ALX dose on diabetic gastroparesis rat model were decisive. The order of the importance was as follows: modeling time > dose of ALX > feed types. Conclusion: The optimized test method can effectively improve the diabetic gastroparesis rat model and provide a technical platform for the study on drug evaluation of diabetic gastroparesis.