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ObjectiveA rapid method for identification of chemical constituents in Puerariae Lobatae Radix dispensing granules was established in order to clarify the material basis. MethodThe chemical constituents of Puerariae Lobatae Radix dispensing granules was qualitatively analyzed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） under positive and negative ion modes， and the chromatographic conditions were on an ACQUITY UPLC HSS T3 column（2.1 mm×100 mm， 1.8 μm） with 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） as mobile phase for gradient elution（0-4 min， 5%-10%B； 4-10 min， 10%-15%B； 10-20 min， 15%-16%B； 20-27 min， 16%-31%B； 27-33 min， 31%-59%B； 33-42 min， 59%-95%B； 42-42.1 min， 95%-5%B； 42.1-45 min， 5%B）， the flow rate was 0.35 mL·min-1， the column temperature was 40 ℃， the injection volume was 5 μL， and electrospray ionization（ESI） was selected. Then these chemical constituents were comprehensively identified based on PeakView 1.2， PubChem， ChemicalBook， ChemSpider， comparative control profiles and literature information. ResultA total of 128 chemical constituents were identified from the dispensing granules， including 60 flavonoids， 26 organic acids， 7 glycosides， 6 coumarins， 3 nucleosides and 26 other compounds. By focusing on the cleavage patterns of flavonoids， organic acids， glycosides， coumarins， nucleosides and other compounds， 12 compounds that have not been reported in Puerariae Lobatae Radix species were identified from the dispensing granules. ConclusionThe established method can systematically and rapidly identify the chemical constituents in Puerariae Lobatae Radix dispensing granules， and cleared it composition is mainly flavonoids and organic acids. Laying a foundation for the study of the material basis， mechanism of action and clinical application of the dispensing granules.
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OBJECTIVE To prepare anemoside B4 （AB4） and programmed cell death ligand 1 （PDL1） siRNA （siP） co- delivered cRGD-modified targeting liposomes （AB4/siP-c-L）， and to study the cellular uptake in vitro. METHODS The cRGD- modified AB4-loaded targeted liposomes （AB4-c-L） were prepared by ethanol injection. AB4-c-L was mixed with 20 nmol/L siP in the same volume and AB4/siP-c-L was obtained through electrostatic adsorption. The particle size， Zeta potential， morphology， encapsulation efficiency and drug content， in vitro release behavior and serum stability of AB4/siP-c-L were investigated by laser scattering particle size tester， transmission electron microscopy， ultrafiltration centrifugation， dialysis and agar-gel electrophoresis block test. Cellular uptake of AB4/siP-c-L by Lewis lung cancer cells LLC and its intracellular localization were evaluated by flow cytometry and confocal laser scan technique. RESULTS The average particle size of AB4/siP-c-L was （187.4±3.1） nm， and the Zeta potential was （33.5±1.4） mV. AB4/siP-c-L was spheroidal in shape. The encapsulation efficiency and content of AB4 were （95.2±0.4） % and （1.0±0.2） mg/mL， respectively. AB4/siP-c-L could better package siP， and exhibited good serum stability， obvious pH sensitivity and sustained release property. The uptake rate of AB4/siP-c-L by LLC cells was significantly higher than that of free drug， and was able to accumulate in cytoplasm. CONCLUSIONS AB4/siP-c-L can effectively realize the co-loading of AB4 and gene drug siP， which has certain in vitro targeting to LLC cells.
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Alismatis Rhizoma(AR)is widely used in Chinese medicine,and its major bioactive components,tri-terpenes,reportedly possess various pharmacological activities.Therefore,it is very important to study the metabolism of triterpenes in vivo.However,the metabolism of AR triterpene extract has not been comprehensively elucidated due to its complex chemical components and metabolic pathways.In this study,an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method,which was based on the characteristic ions from an established database of known triterpenes,was used to analyze the major metabolites in rats following the oral administration of Alismatis Rhizoma extracts(ARE).As a result,a total of 233 constituents,with 85 prototype compounds and 148 metabo-lites,were identified for the first time.Hydrogenation,oxidation,sulfate and glucuronidation conjugation were the major metabolic pathways for triterpenes in AR.In addition,the mutual in vivo transformation of known ARE triterpenes was discovered and confirmed for the first time.Those results provide comprehensive insights into the metabolism of AR in vivo,which will be useful for future studies on its pharmacodynamics and pharmacokinetics.Moreover,this established strategy may be useful in meta-bolic studies of similar compounds.
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Objective@#To optimize the extraction technology for Gegen-Shujin granules.@*Methods@#With yield of volatile oil as index, single factor tests were used to investigate effects of water, soaking time and distillation time on extraction technology of volatile oil. The water amount, extraction time and numbers of extraction as factors, the contents of puerarin and total solid as indexes, orthogonal test was employed to optimize the extraction technology of Gegen-Shujin granules.@*Results@#The optimical extraction technology conditions were as follows: Cinnamomi Ramulus, cinnamomi ramulus, notopterygium, turmeric were extracted to get volatile oil with eight-folds amount water of herbs for 6 hours; while the other herbs were boiled with ten-folds amount water of herbs and extrancted for two times, 1.5 hours each time.@*Conclusions@#This extraction process is reasonable and practical, and can guarantee the quality of preparation.
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Objective To optimize the extraction technology for Gegen-Shujin granules. Methods With yield of volatile oil as index, single factor tests were used to investigate effects of water, soaking time and distillation time on extraction technology of volatile oil. The water amount, extraction time and numbers of extraction as factors, the contents of puerarin and total solid as indexes, orthogonal test was employed to optimize the extraction technology of Gegen-Shujin granules. Results The optimical extraction technology conditions were as follows: Cinnamomi Ramulus, cinnamomi ramulus, notopterygium, turmeric were extracted to get volatile oil with eight-folds amount water of herbs for 6 hours; while the other herbs were boiled with ten-folds amount water of herbs and extrancted for two times, 1.5 hours each time. Conclusions This extraction process is reasonable and practical, and can guarantee the quality of preparation.
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AIM:To investigate the antagonism mechanism of benazepril on renal fibrosis of unilateral uretreal obstructire (UUO).METHODS:Fiftyfour Sprague-Dawley (SD) rats were randomly divided into three groups:sham operation group,model group,benazepril group.All the other groups were treated with surgery method of unilateral ureteral ligation to establish a rat model of renal fibrosis except the sham operation group.The pathological changes were observed by Masson staining;the positive staining expression of TGF-β1,Col-Ⅳ,Wnt1,Wnt4 in frozen sections were observed by immunofluorescence staining;the expressions TGF-β1,Col-Ⅳ,Wnt1 and Wnt4 mRNA in renal tissues were detected by real time fluorescence quantitative PCR.RESULTS:Masson staining results:compared with the sham group,the parts of blue dye collagen fiber and the degree of renal fibrosis increased significantly in all modeled groups;compared with the model group,the parts of blue dye collagen fiber and the degree of renal fibrosis decreased significantly in benazepril group.Immunofluorescence and PCR results:compared with the model group,the positive expression of Wnt1,Wnt4,TGF-β1,Col-Ⅳ and mRNA in the benazepril group decreased significantly.CONCLUSION:Benazepril can improve renal fibrosis by inhibiting the expression of TGF-β1,Col-Ⅴ,Wnt1,Wnt4 and ameliorating the level of renal fibrosis.
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Background and purpose: It was reported that activins, members of transforming growth factor-β superfamily, could induce several tumor cells into apoptosis. This study was designed to observe the effects of recombination human activin A on proliferation and apoptosis ofA549 cells. Methods: A549 cells were cultivated by routine method, then treated with different concentrations of recombination human activin A. Inhibitory effect of activin A on A549 cell proliferation was detected by MTT assay, and apoptosis of A549 cells was determined by flow cytometry. The expression of type Ⅱ receptors of activin (ActR Ⅱ and ActR Ⅱ B) was detected by Western blot. Results: Recombination human activin A inhibited the proliferation ofA549 cells in time-dependent and dose-dependent manners. Increases of apoptosis in A549 cells and expression of ActR I] and ActR ⅡB occurred dose-dependently after treated with acfivin A. Conclusion: Recombination human activin A inhibited cell proliferation and induced apoptosis in A549 cells, at least in part, by inducing expression of type Ⅱ receptors of activin and activation of its downstream signaling.
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OBJECTIVE@#To explore the characteristic of formant-a very important parameter in the spectrogram of three types of artistic voice (western mode; Chinese mode; Beijing opera).@*METHOD@#We used MATLAB software to make the short-time Fourier transform and spectrogram analysis on the homeostasis vowel examples of the three types.@*RESULT@#The western mode had different representation "singer formant" (Fs) based on the voice part; the Chinese mode's notable features were that F1, F2, F3, were continuous and the energy of them changed softly; the Beijing opera had the common representation which was a very wide formant and there was soft transition between formants and various harmonic, besides it showed a similar component like the "Fs" (two formants connected normally).@*CONCLUSION@#Different artistic voice showed their own characteristics of the formant parameter in the spectrogram, which had important value on the identification, objective evaluation and prediction.
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Adult , Female , Humans , Male , Middle Aged , Young Adult , Music , Spectroscopy, Fourier Transform Infrared , Voice QualityABSTRACT
Objective To study expression of hypoxia inducible factor 1? (HIF 1?) in the lungs of hypoxic rats and to explore the role of HIF 1? in the pathogenesis of hypoxic pulmonary hypertension Methods Twenty Wistar rats were randomly divided into control group and hypoxia group The models of hypoxic pulmonary hypertension were established by exposure to hypoxia for 4 weeks according to our laboratory protocol Digoxin labelled cRNA probe for HIF 1? was prepared by in vitro transcription Northern blot was performed by using the HIF 1? cRNA probe and In situ hybridization was also conducted with rat lung tissue sections Results Northern blot hybridization showed minimally positive in the lung tissue of control group ,but strongly positive in hypoxia group In situ hybridization analysis with hypoxic rat lung tissue revealed that HIF 1? mRNA was expressed in bronchial epithelial cells (strongly positive reaction) and peribronchial proliferative lymphomatic tissue (weakly positive reaction),casually in alveolar wall cells (weakly positive reaction),but not in pulmonary arterial endothelium and smooth muscle cells Conclusions Our data suggested that chronic hypoxia could induce pulmonary hypertension characterized by pulmonary vascular remodeling HIF 1? mRNA expression was elevated in the lungs of rat under hypoxic condition HIF 1 may be involved in the pathophysiological changes in response to hypoxia and play an important role in the development of hypoxic pulmonary hypertension