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ObjectiveTo clone squalene epoxidase (SE), a potential key rate-limiting enzyme involved in the synthesis pathway of Poria cocos triterpenes, from P. cocos and analyze for bioinformatics and expression. MethodThe total RNA was extracted by the kit and reverse-transcribed to cDNA. Specific primers were designed, and the cDNA was used as a template for cloning the SE gene, which was analyzed for bioinformatics. The expression of P. cocos qualene epoxidase(PcSE) was examined by Real-time polymerase chain reaction(Real-time PCR) in P. coco Shenzhou No. 10, Xiangjing 28, and 5.78 strains. ResultThe full length of PcSE is 1 571 bp, containing four exons and three introns. The obtained CDS sequence is 1 413 bp, encoding 470 amino acids. This protein is a hydrophobic protein with no signal peptide structure and has two transmembrane structural domains with a FAD/NAD (P) binding domain and SE structural domain localized to the mitochondrial membrane and the plasma membrane. The homologous sequence alignment with fungi of the Poriferae family is 80.92%, and the phylogenetic tree shows that PcSE protein is most closely related to P. cocos from the US. The results of Real-time PCR showed that the PcSE was expressed in all three strains, with the highest expression in 5.78 strain, and there was no significant difference in PcSE expression among the three strains. ConclusionFor the first time, the PcSE gene was cloned and analyzed from P. cocos, providing a basis for further research on the function of PcSE and the analysis of P. cocos triterpene biosynthesis pathway.
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OBJECTIVE To optimize the extraction process and to primarily evaluate the anti-anxiety and anti-depression efficacy of polysaccharide from Baihe dihuang decoction. METHODS Based on Plackett-Burman experimental design, using the comprehensive score of yield and content of polysaccharide as indicators, with extraction time, water amount, alcohol precipitation concentration as factors, Box-Behnken response surface methodology was used to optimize the extraction process of polysaccharide from Baihe dihuang decoction; and the validation test was conducted. Forty ICR mice were divided into control group, venlafaxine group [positive control, 13.5 mg/(kg·d)], Baihe dihuang polysaccharide high-dose and low-dose groups [5.28, 2.64 g/(kg·d),by raw material], with 10 mice in each group (half male and half female). Administration groups were given corresponding drug solution intragastrically, and control group was given water 10 mL/kg intragastrically, once a day, for 7 test, forced swimming test and tail suspension test were used to evaluate the effects of the extract prepared by the optimal process on the anxiety-like and depression-like behavior of mice; enzyme-linked immunosorbent assay was used to detect the effects of the extract on the levels of neurotransmitter in cerebral tissue of mice. RESULTS The optimal extraction process of Baihe dihuang decoction was: the water amount of 25 times, extract time of 1.5 hours, and alcohol precipitation concentration of 70%. In 3 times of validation test, the average yield and content of polysaccharide were 33.10% and 0.62 mg/mg, the relative deviations of which from the predicted values (36.14% and 0.65 mg/mg) were 8.40% and 4.62% respectively (RSD<2%, n=3). The polysaccharide extract of Baihe dihuang decoction could effectively increase the percentages of open-arms entry, the percentages of open-arms time, the total distance of voluntary activities and the activity distance in central area, and significantly shortened the immobility time of forced swimming test and tail suspension test (P<0.05 or P<0.01). The polysaccharide extract could significantly increase the levels of 5-hydroxytryptamine, norepinephrine (except for the Baihe dihuang polysaccharide low-dose group) and gamma-aminobutyric acid in cerebral tissue of mice, while significantly decrease the levels of glutamic acid (except for the Baihe dihuang polysaccharide low-dose group) (P<0.05 or P< 0.01). CONCLUSIONS The optimized extraction process of polysaccharide from Baihe dihuang decoction is stable and feasible, and the obtained polysaccharide extract has obvious anti-anxiety and anti-depression effect in vivo.
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Through the textual research and analysis of the variety, origin, processing, quality evaluation and clinical application of Moslae Herba in ancient and modern literature, its origin of materia medica was clarified. Moslae Herba has experienced variety changes in history. Elsholtzia ciliata was the mainstream variety during and before the Song dynasty, however, during the Ming and Qing dynasties, emerging variety of Mosla chinensis rose to the mainstream status due to its remarkable efficacy and the formation of cultivation, and differentiated into two commodities(wild variety of Qingxiangru and cultivated variety of Jiangxiangru), cultivated products formed an authentic producing area in Jiangxi. The three varieties coexisted during the Ming and Qing dynasties, and the Elsholtzia varieties were gradually eliminated. Variety changes have caused changes in the functions and indications of drugs. E. ciliata had the effect of clearing heat and was mainly used to treat heatstroke and cholera, while M. chinensis was used for exogenous wind cold and dampness in the summer because of its warm and strong sweating properties, but not for cholera. Traditional Moslae Herba is mainly harvested in the summer and autumn (flowering to fruiting stage) and the above-ground parts are dry in the shade and used as medicine. Modern Qingxiangru is mostly harvested before the flowering period, and Jiangxiangru is harvested after flowering and fruiting in late summer and early autumn. In summary, according to the 2020 edition of Chinese Pharmacopoeia, the dried above-ground parts of Moslae Herba should be selected for Xinjia Xiangruyin in the Catalogue of Ancient Famous Classical Formulas(The First Batch), mainly the cultivated variety of Jiangxiangru, and the raw products is cut into segments and used as medicine. It is suggested that when applying and developing famous classical formulas containing Moslae Herba at different periods of time today, the origin should be established in conjunction with clinical efficacy.
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Through the textual research and analysis of ancient and modern documents of Acanthopanacis Cortex(AC), this paper combed the variety, origin, harvesting, processing and ancient quality evaluation methods of AC, and clarified the historical context of the mixing of its common counterfeit product(Periplocae Cortex), in order to provide a basis for the development of famous classical formulas containing AC. AC was first published in Shennong Bencaojing with the name of Wujia, Wujiapi is the name rectification in all dynasties since Leigong Paozhilun. According to the description of inflorescence location and fruit morphology of Wujia in the materia medica, it is judged that the mainstream origin of AC used in previous dynasties was Acanthopanax gracilistylus. Periplocae Cortex was mixed with AC in the period of the Republic of China because it was in line with the "like Lycii Cortex, light, brittle and fragrant". The origin of Wujiapi recorded in past dynasties was concentrated in the middle and lower reaches of the Yangtze River, mainly in Hubei, Henan, Anhui and other places. Since modern times, the traditional quality evaluation of AC has been gradually summarized, with thick skin, white color and fragrant smell as the best. The traditional harvesting and processing of AC involved picking the stems in May and July of the lunar calendar, picking the roots in October, and drying in the shade. In modern times, the roots of AC are harvested, washed, peeled and dried in summer and autumn. In the past dynasties, there were rice wine processing, Euodiae Fructus boiling, ginger juice processing and other methods. In modern times, it is usually cut into thick slices after the cleansing. According to the research results, it is suggested that the root bark of A. gracilistylus should be selected as the origin of AC in famous classical formulas, which should be processed into the medicine according to the specific prescription requirements. In addition, it is suggested to restore the medicinal name of Periplocae Cortex as Yangtao, in order to reduce its chaotic influence on the medicinal use of AC.
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Objective To determine the dissolution rate of Erzhi pill which was prepared by using different crushing technologies with specnuezhenide as index , and study the influence of superfine pulverization technology on the dissolution of Erzhi pill. Methods Crude drugs were crushed into the fine powder by employing general crusher and ultra-micro pulverizer,respectively. The water-honeyed pills were prepared by using these fine powders. The method of rotating basket and HPLC were used for the determination of dissolution of specnuezhenide in vitro. The data were analyzed by applying the law of Weibull. Results Compared with Erzhi pill B,the dissolution parameters of specnuezhenide in Erzhi pill A ,including T50、T60、T70、T80、T90、F1h and F2h, showed significant differentce(P<0.05). Conclusion Applying the superfine pulverization technology can improve the dissolution of Erzhi pills in vitro.
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Objective To optimize the molding process of Erzhi pill. Methods Using the external properties of pill, the pill weight, disintegration time as the comprehensive evaluation index, the L9(34) orthogonal test was used to test the effects of three factors:the relative density of thick paste(A), the powder fineness(B), and the proportion of powder and honey(C). Results The best molding process was as follows: the relative density of thick paste was 1.25, the powder fineness was 100 meshes, and the proportion of powder and honey was 1: 0.6. Conclusion This optimized molding process proved to be repeatable, simple and feasible. The Erzhi pill made by this preparation technology are round, the color and luster are uniform and the dissolving time is short. It provides the basis for studying the molding process of Erzhi pill .
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<p><b>OBJECTIVE</b>To establish the double HPLC fingerprints of water-soluble composition and amino acids precolumn derivative reagent of 13 batches of Isatidis Radix micropower.</p><p><b>METHOD</b>The gradient elution was adopted with Hypersil BDS C18 column (4.6 mm x 250 mm, 5 microm). Water-soluble ingredients were detected with acetonitrile-0.1% phosphoric acid solution as mobile phase, flow rate 0.5 mL x min(-1), column temperature 20 degrees C, and the injection volume 10 microL. Amino acid ingredient were derived by PITC, and then were detected with mobile phase of 0.1 mol x L(-1) sodium acetate buffer solution (pH 6.5) - acetonitrile, flow rate 1.0 mL x min(-1), column temperature 30 degrees C, and the injection volume 5 microL. Both of the absorption wavelengths were 254 nm. Pharmacopoeia Commission "Chinese chromatographic fingerprint evaluation system (version 2.0)" was adopted to analyse, fingerprints of Isatidis Radix micropower were established, at the same time 4 main ingredients were recognized by the SPSS cluster analysis.</p><p><b>RESULT</b>Common mode of Fingerprint to water-soluble and amino acids ingredient of Isatidis Radix micropower was established, then adenosine, epigoitrin and 15 amino acids were identified as characteristic peaks. Cluster analysis showed that different kinds of the herbal Isatidis Radix micropower from different areas were different levels in the main ingredients.</p><p><b>CONCLUSION</b>Double fingerprints of Isatidis Radix micropower is established. Each peak is optimally separated in chromatogram, which provides a scientific basis for quality control of Isatidis Radix micropower.</p>
Sujets)
Acides aminés , Isatis , Chimie , Poudres , Contrôle de qualitéRésumé
AIM: To investigate the influence of microwave-assisted extraction on flavone contents of Pollen Tyhae micropowder. METHODS: Ultraviolet spectrophotometer and HPLC were applied to analyze Pollen Tyhae micropowder, total flavon and isorhamnetin-3-O-neohespridoside were adopted as the marker, respectively. RESULTS: Appropriate conditions of microwave-assisted extraction included: extraction time of 8min, ethanol concentration of 70%, Solid/liquid ratio of 1∶18 (g?mL -1) and the power of microwave oven of 540w. CONCLUSION: Compared with normal reflux method, microwave-assisted extraction of Pollen Typhae micropowder is more useful and can improve the extraction rate the reduce the extracting time.
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AIM: To provide a HPLC fingerprint of Fructus aurantii micropowder in order to create a basis for(identification.) METHODS: With the help of computer similarity evaluation,Seventeen kinds of Fructus aruantii sample were pulverized and micronized separately,then were extracted by methanol and compared correlation between both extracts. RESULTS: In methanol extraction,contents of aurantiamarin,hesperidin and neohesperidin in micropowder were more than that in pulverized powders,correlation of both HPLC fingerprint was 0.9~1.0. CONCLUSION: The method established can be used for identifying and evaluating crud drug of Fructus aurantii,and further prove that micronization is beneficial to increasing flavonoid dissolution.
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AIM: To provide application and identification basis for methanol-extratis of Fructus aurantii by constructing the HPLC-FP. METHODS: The C_(18) column was used with a mobile phase of(A)(0.095%) phosphoric acid acetonitrile-(B)(0.095%) phosphoric acid gradient elution.The flow rate was(1.0) mL/min.The wavelength of detecter was set at 330 nm.Naringin was reference standard. RESULTS: By cluster analysis,the eighteen kinds of Fructus aurantii samples were classified as four clusters: the superior in producing area,the ordinary in producing area,the ordinary and the inferior.By similarity calculation,the similarity of the superior in producing area and the ordinary in producing area were(0.9)~(1.0),and the ordinary and the inferior were less than(0.9). CONCLUSION: The HPLC-FP of Fructus aurantii has been established.The method can be used to identify and evaluate the quality of Fructus aurantii.
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Objective: To select the optimum alcohol concentration of Shangtong Tincture. Methods: The percutaneous absorption test was performed on excised rats skin in improved Franz diffusion cell. The penetrative amount of paeonol was determined quantitatively by HPLC and the content of paeonol as a marker. Results: The penetrative amount of paeonol of Shangtong Tincture with 60%~80% alcohol concentration have a optimal value, the average rate is 0.010mg/cm 2?h. Conclusion: Shangtong Tincture with 70% alcohol concentration is the best.
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Objective: To provide experiment data for ultra fine prepared mineral drugs Methods: Electron microscope scannin,X ray diffraction were used in identification, atomic emit sectrum was used in determination. Results: The dissolution rate of ca 2+ composition could be high.The ultra fine prepared of minaral drugs could be prepared with powder diameter of K 4。 Conclusion: Ultra fine Prepared fluoritum and gypsum fibrosum may Save clinically dose.