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ObjectiveTo screen and validate key enzyme genes affecting the polysaccharide content in different Polygonatum species and perform in-depth amino acid sequence analysis by transcriptomic analysis of P. zanlanscianense， P. kingianum， and P. cyrtonema rhizomes to enrich the transcriptome data of Polygonatum plants and provide references for polysaccharide biosynthesis mechanism and genetic improvement. MethodThe Polygonatum transcriptome was sequenced and analyzed using the Illumina NovaSeq high-throughput sequencing platform， and the differences in the transcriptomes of the three Polygonatum species were compared and according to the annotations of Nr， Gene Ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） databases. The key enzymes in the polysaccharide metabolism pathway were screened， and the expression of key enzyme genes was clustered and correlated with the polysaccharide content. Finally， Real-time polymerase chain reaction （Real-time PCR） was performed to validate the eight key enzyme genes， and the key genes of polysaccharide biosynthesis were further screened for homologous gene sequence analysis in combination with sequencing results， followed by constructing phylogenetic trees， predicting motifs， conserved structural domains， protein sequence isoelectric points， and molecular weights， and constructing 3D protein structures by using homology modeling method. ResultThe annotation of the Nr database revealed that three Polygonatum species had the highest gene homology with Asparagus officinalis. GO database annotation results showed that three Polygonatum species differed significantly in binding， catalytic activity， metabolic processes， and cellular components， while the KEGG pathway annotation results indicated that three Polygonatum species differed significantly in the starch and sucrose metabolic pathway and galactose metabolic pathway. According to clustering analysis， correlation analysis， Real-time PCR， expression profiles， and structural and functional predictions of amino acid sequences， the key enzyme significantly affecting the polysaccharide content in different Polygonatum species was inferred to be β-fructofuranosidase （sacA）. ConclusionSacA may be the main influencing factor for the difference in polysaccharide content of Polygonatum， and is also an important reason why Polygonatum polysaccharides are mainly fructans.
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OBJECTIVE：To compare th e difference of the components of volatile oil in Citrus medica from different producing areas. METHODS ：The volatile oil of C. medica from 10 different producing areas was extracted with steam distillation ，and the yield was calculated. The components of the volatile oil of C. medica from different producing areas were analyzed by GC-MS. The compounds were retrieved from NIST 14.L mass spectrum database and identified. Relative mass fraction of chemical component was determined by peak area normalization method. Cluster analysis of samples were performed by using SPSS 20.0 software. RESULTS：The yields of volatile oil of C. medica from 10 different producing areas were 0.10％-1.75％，among which sample from Qianwei county in Leshan city of Sichuan province was the highest （1.75％）. A total of 66 components were identified in the volatile oil of C. medica from different producing areas ，with a relative molecular weight of 126.20-392.66. The majority was C 10 and C 15 compounds；isomers with relative molecular weight of 136，154 took up the great proportion ，which were mainly cycloalkane monoterpenes. There were 12 common components in the volatile oil of C. medica from different areas ，which were limonene（24.90％），terpinene（14.71％），（－）-4 terpineol（2.88％），citral（2.33％），α-myrrhene（2.33％），geraniol（1.52％）， α-pinene（1.37％），trans bergamot olene （1.16％），isoterpinene（1.13％），methyl palmitate （1.12％），linalool（1.09％）and geranyl acetate（1.04％）according to relative mass fraction ；8 of them were monoterpenes ，2 were sesquiterpenes and 2 were esters. There were 4 categories of C. medica from different producing areas ，i.e. S 2，S4，S6 clustered into one ；S1，S3，S7，S8 clustered into one ； S5 and S 10 clustered into one ；S9 as one . CONCLUSIONS ： There are some difference of the components in volatile oil of medica from different producing areas ，and the content of the same component also has great difference in the volatile oil of C. medica from different producing areas.
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Objective::To analyze and identify the chemical constituents of Citri Sarcodactylis Fructus by an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbirap HRMS) method. Method::Compounds were separated on Thermo Scientific Accucore™ C18 column (3 mm×100 mm, 2.6 μm). The mobile phase was 0.1% formic acid solution and 0.1% formic acetonitrile solution. The flow rate was 0.3 mL·min-1, and the column temperature was set at 30 ℃. HRMS was performed using an electrospray ion source (ESI), and scanned in a positive ion mode by means of full scan/data dependent secondary scan (Full MS/dd-MS2). Compound Discoverer 3.0 software that could be linked to mzCloud network database and local high-resolution mass spectrometry database of traditional Chinese medicine components was used to analyze the data, based on accurate molecular mass, retention behaviors and characteristic ion fragmentation of the compounds, as well as literature information and relevant reference materials. Result::Totally 54 chemical constituents in Citri Sarcodactylis Fructus were identified, including 16 flavonoids, 17 coumarins, 3 limonoids, 6 nucleosides and nucleobases, 2 organic acids, 3 aromatic aldehydes, 1 amino acid and 6 other components. Conclusion::The established UPLC-Q-Orbitrap HRMS method can be used to effectively and rapidly identify main chemical constituents of Citri Sarcodactylis Fructus. The findings provide a methodological reference and theoretical foundation for defining the pharmacodynamic material base and optimizing quality control index of Citri Sarcodactylis Fructus, which is of guiding significance for further development and utilization of the resources.
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OBJECTIVE： To optimize the purification technology of total flavonoids from Sparganium stoloniferum. METHODS： Separation and purification by macroporus adsorption resin， using sample solution pH， flow rate and concentration of eluent， the purification rate of total flavonoids as evalution indexes， the purification technology of total flavonoids from S. stoloniferum were optimized by Box-Behnken design-response surface methodology based on single factor test. Validation test was conducted. RESULTS： The optimal purification technology was sample solution pH 4.8， flow rate of eluent 2.0 BV/h， concentration of eluent 72％. The purification rate of total flavonoids in 3 batches of samples was 72.34％ （RSD＝1.77％， n＝3） in validation test， relative errors of which to predicted value （73.99％） was 2.13％. CONCLUSIONS： The optimal purification technology is stable and feasible， and can be used for the purification of total flavonoids from S. stoloniferum.
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Abstract Background: The cardiac hypertrophy (CH) model for mice has been widely used, thereby providing an effective research foundation for CH exploration. Objective: To research the effects of CH modeling under abdominal aortic constriction (AAC) using different needles and weights in mice. Methods: Four needles with different external diameters (0.35, 0.40, 0.45, and 0.50 mm) were used for AAC. 150 male C57BL/6 mice were selected according to body weight (BW) and divided into 3 weight levels: 18 g, 22 g, and 26 g (n = 50 in each group). All weight levels were divided into 5 groups: a sham group (n = 10) and 4 AAC groups using 4 ligation intensities (n = 10 per group). After surgery, survival rates were recorded, echocardiography was performed, hearts were dissected and used for histological detection, and data were statistically analyzed, P < 0.05 was considered statistically significant. Results: All mice died in the following AAC groups: 18g/0.35 mm, 22 g/0.35 mm, 26 g/0.35 mm, 22 g/0.40 mm, and 26 g/0.40 mm. All mice with AAC, those ligated with a 0.50-mm needle, and those that underwent sham operation survived. Different death rates occurred in the following AAC groups: 18 g/0.40 mm, 18 g/0.45 mm, 18 g/0.50 mm, 22 g/45 mm, 22 g/0.50 mm, 26 g/0.45 mm, and 26 g/0.50 mm. The heart weight/body weight ratios (5.39 ± 0.85, 6.41 ± 0.68, 4.67 ± 0.37, 5.22 ± 0.42, 4.23 ± 0.28, 5.41 ± 0.14, and 4.02 ± 0.13) were significantly increased compared with those of the sham groups for mice with the same weight levels. Conclusion: A 0.45-mm needle led to more obvious CH than did 0.40-mm and 0.50-mm needles and caused extraordinary CH in 18-g mice.
Resumo Fundamentos: O modelo de hipertrofia cardíaca (HC) para ratos foi amplamente utilizado, proporcionando assim uma base de pesquisa efetiva para a exploração de HC. Objetivo: pesquisar os efeitos do modelamento de HC sob constrição da aorta abdominal (CAA) usando diferentes agulhas e pesos em ratos. Métodos: foram utilizadas quatro agulhas com diâmetros externos diferentes (0,35, 0,40, 0,45 e 0,50 mm) para CAA. Foram selecionados 150 ratos C57BL / 6 machos de acordo com o peso corporal (PC) e divididos em 3 níveis de peso: 18 g, 22 g e 26g (n = 50 em cada grupo). Todos os níveis de peso foram divididos em 5 grupos: um grupo sham (n = 10) e 4 grupos CAA usando 4 intensidades de ligadura (n = 10 por grupo). Após a cirurgia, foram registradas as taxas de sobrevivência, foi realizada ecocardiografia, os corações foram dissecados e utilizados para detecção histológica, e os dados foram analisados estatisticamente, p < 0,05 foi considerado estatisticamente significante. Resultados: Todos os ratos morreram nos seguintes grupos de CAA: 18 g/0,35 mm, 22 g/0,35 mm, 26 g/0,35 mm, 22 g/0,40 mm e 26 g/0,40 mm. Todos os ratos com CAA, aqueles ligados com uma agulha de 0,50 mm, e aqueles que sofreram operação sham sobreviveram. Ocorreram diferentes taxas de mortalidade nos seguintes grupos de CAA: 18 g/0,40 mm, 18 g/0,45 mm, 18 g/0,50 mm, 22 g/45 mm, 22 g/0,50 mm, 26 g/0,45 mm e 26 g/0,50 mm. As proporções de peso do coração/peso corporal (5,39 ± 0,85, 6,41 ± 0,68, 4,67 ± 0,37, 5,22 ± 0,42, 4,23 ± 0,28, 5,41 ± 0,14 e 4,02 ± 0,13) aumentaram significativamente em comparação com os grupos sham para ratos com os mesmos níveis de peso. Conclusão: uma agulha de 0,45mm levou a HC mais óbvia do que as agulhas de 0,40 mm e 0,50mm e causou HC extraordinária em ratos de 18 g.
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Animals , Male , Aorta, Abdominal , Body Weight , Cardiomegaly/etiology , Disease Models, Animal , Needles/standards , Reference Values , Time Factors , Echocardiography , Random Allocation , Reproducibility of Results , Cardiomegaly/pathology , Constriction , Ligation/instrumentation , Mice, Inbred C57BLABSTRACT
This study was aimed to observe the effect of Bai-Zhu Fu-Ling (BZFL) Decoction in different proportion-ing on VIP and VIPR1 in Crohn's disease (CD) rats with spleen deficiency syndrome, in order to further explore the immunologic mechanism of BZFL Decoction on CD. The CD rat model with spleen deficiency syndrome was estab-lished using exhaustion and hunger. The model rats were treated by BZFL Decoction with different proportioning, and immunohistochemistry (IHC) was used to detect the expression of VIP and its receptor in colon tissues. The results showed that comparing to the blank control group, the level of VIP and its receptor of the model group significantly increased (P< 0.05). Comparing to the model group, the level of VIP and its receptor in BZFL Decoction B5 group (Rhizoma A tractylodis Macrocephalae:Poria = 12:15), B6 group (Rhizoma A tractylodis Macrocephalae:Poria = 15:12) and B7 group (Rhizoma A tractylodis Macrocephalae:Poria = 18:9) was significantly decreased (P< 0.05). It was con-cluded that the effect of BZFL Decoction of B5 group, B6 group and B7 group was better than other groups in VIP and its receptor which can regulate the VIP and its receptor, inhibit the releasing of inflammatory factors and reduce intestinal inflammation injury.
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s: The onset of Crohn’s disease (CD) is the interaction of environment, heredity, infection, immune and other factors. It is also closely related to abnormal immune functions. Without special treatment, CD is identified as a modern refractory disease. By syndrome differentiation and treatment, traditional Chinese medicine (TCM) can effectively relieve disease conditions, improve the quality of life and reduce side effects of modern medication. The core compatibility ofBai-Zhu andFu-Ling can reinforce spleen-qi and dispel dampness, which met the common pathogenesis of CD. Therefore, the combination is comprehensively used in the compound prescription. Our previous study found thatBai-Zhu Fu-Ling decoctioncan reduce the vasoactive intestinal peptide (VIP) of animal model of spleen-qi deficiency, downregulate VIP receptors, decrease the affinity of VIP receptors and improve animal model’s sIgA. To further clarify the effects about neurotransmitters and their correlation with the immune system in the pathogenesis of CD and the intervention mechanism treated by different proportional compatibility ofBai-Zhu Fu-Ling decoction, we studied influences of the decoction on related transmitters of nerve- immune network and functions of receptors, as well as cytokine secretion and signal transduction of TLR4-NF-κB. Our studies can provide references and foundations to further explore TCM treatment of CD.
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Aim: To provide the foundation for reasonable utilization by analyzing the essential oils of Sabina pingii var. wilsonii.. Methods: The essential oils were extracted using steam distillation and separated with GC capillary column. The components were quantitatively determined with normalization method, and were preliminarily identi-fied by GC-MS. Results: 79 Components were identified, which took up 93. 78% of the essential oils. Conclusion: The main components of essential oils were a-terpineol( 16. 70%); α-fenchene( 14. 98%); 2,6,10-dodecatrien-1-ol, 3,7, 11-trimethyl-( 6.49%); 1, 4-cyclohexadiene, 1-methyl-4-[1-methylethyl]-( 4. 74%); kaura-5, 16-dien-18 [orl9]-ol(3. 62%); naphthalenemethanol, 1, 2, 3, 4, 4a, 5, 6, 7-octahydro-α, α, 4a, 8-tetramethyl-, [2R-cis]-(3. 04%); [ + ]-4-carene ( 2. 81%); D-limonene ( 2. 71%); β-myrcene ( 2. 59%); cedrol (2.40%); a-pinene (2. 36%); 7-isopropyl-1, 1, 4a-trimethyl-1 ,2,3,4,4a, 9,10,10a-octahydrophenanthrene(2. 32%).
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OBJECTIVE: To establish the quality standard of Shujinzhuanggu pill. METHODS: Thin-layer chromatography(TLC) was employed for the qualitative determination of Psoralea corylifolia,Panax notoginseng,Angelica sinensis, Cyperus rotundus and Dipsacus asperoides. High performance liquid chromatography(HPLC) was applied for the determination of Psoralen in psoralea corylifolia in Shujin zhuanggu pill. RESULTS: The characteristic identification by TLC was distinctive and highly specific. The linear range of Psoralen was 0.04～0.28 ?g (r=0.999 2). The average recovery was 98.84 and RSD was 2.17%. CONCLUSION:The established standard can be used for the quality control of Shujin zhuanggu pill.
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Objective To study their chemical components of xianglingcao(Crotalaria ferruginea) and identify their chemical structures.Methods The compounds were isolated by chromatography and their structures were identified by spectral analysis and compared with the published data.Results Ten compounds were isolated and identified as vomifolilol(Ⅰ),genistein(Ⅱ),p-hydroxybenzoic acid(Ⅲ),5,7-dihydroxy-4-methoxy-flavone-7-O-?-D-glycopyranoside(Ⅳ),octadecoic acid(Ⅴ),octacosanol(Ⅵ),?-stitosterol(Ⅶ),?-daucosterol(Ⅷ),stigmasterol(Ⅸ),and ?5,22 stigmasterol-3-O-?-D-glycopyranoside(Ⅹ).Conclusion The ten compounds are obtained from xianglingcao for the first time.