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BACKGROUND:Osteonecrosis of the femoral head is a common and disabling disease,which is mainly characterized by microcirculation disorders and bone cell metabolism disorders.Luzhongjiangu decoction was developed by Shandong Academy of Chinese Medicine and used in the form of soup in the clinic,which has good efficacy in the treatment of osteonecrosis of the femoral head.However,its mechanism of action has not been clarified. OBJECTIVE:To study the effect mechanism of Luzhongjiangu decoction on intestinal flora in rats with osteonecrosis of the femoral head based on 16S rDNA sequencing technique. METHODS:The model of osteonecrosis of the femoral head was established in Wistar rats by intragastric administration of retinoic acid.The therapeutic effect of Luzhongjiangu decoction was evaluated by serum hormone,bone histopathology and serum hormone levels.16s rDNA sequencing technique was used to detect the intestinal flora of rats in the blank control group,model group and middle-dose Luzhongjiangu decoction group.The corresponding library was constructed and OTU clustering and microbial community diversity and abundance analysis were carried out to determine the composition of intestinal flora and the changes of species and diversity among groups. RESULTS AND CONCLUSION:Luzhongjiangu decoction could significantly increase the expression of osteocalcin,osteopontin and other osteogenic related factors,alleviate the destruction of bone trabeculae,increase bone mineral density,and had a significant therapeutic effect on osteonecrosis of the femoral head,of which the middle dose group showed the most significant effect.The results of intestinal flora sequencing showed that Luzhongjiangu decoction improved the flora disorder of rats with osteonecrosis of the femoral head to some extent,and screened out different colonies such as Bacillus,Desulfurizans,Desulfurization,Isobacteria,Bifidobacterium and so on;it could up-regulate the abundance of beneficial bacteria such as Bifidobacterium,down-regulate the abundance of harmful bacteria such as Desulfovibrio,and improve the structure of intestinal flora.Functional prediction analysis indicated that Luzhongjiangu decoction could mainly affect amino acid metabolism and energy metabolism.Correlation analysis showed that the differential bacteria of Bifidobacterium and Intestinimonas in the middle dose group of Luzhongjiangu decoction were positively correlated with vitamin D3,estradiol and calcitonin,and negatively correlated with prostaglandin E2.In the model group,Escherichia-Shigella,Desulfovibrio,Globicatella and Streptococcus were positively correlated with prostaglandin E2 and negatively correlated with vitamin D3,estradiol and calcitonin.To conclude,Luzhongjiangu decoction may play a role in the treatment of osteonecrosis of the femoral head by regulating the structure of intestinal flora,up-regulating the abundance of beneficial bacteria and affecting the secretion of vitamin D3,estradiol,calcitonin and prostaglandin E2.
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OBJECTIVE To study the effects of Dayuanyin decoction （DYY） on lung injury and gut microbiota in mice with RSV-Hanshi yufei syndrome. METHODS Thirty-six BALB/c mice were divided into the normal group （NC group）， model group （MC group）， positive control group （LBM group， 57.59 mg/kg ribavirin）， DYY low-dose， medium-dose and high-dose groups （LDYY，MDYY，HDYY groups，1.67，3.34，6.68 g/kg，calculated by crude drug）， with 6 mice in each group. Except for NC group， RSV-Hanshi yufei syndrome model was induced by “Hanshi modeling+RSV infection”； after the second cold and wet stimulation， RSV solution was dripped into the nasal cavity at 50% tissue culture infectious dose， once a day， for 3 consecutive days； each administration group was given corresponding solution intragastrically 4 hours after each nasal drip， once a day， for 5 consecutive days. The levels of motilin （MTL） and gastrin （GAS） in serum， the levels of IL-6 and IL-1β in supernatant of lung tissue， and the percentage of peripheral blood lymphocytes were all determined. The pathological changes of lungs were observed， and the changes of gut microbiota were analyzed. RESULTS Compared with MC group， the levels of MTL in serum （except for HDYY group）， and the levels of IL-6 and IL-1β in supernatant of lung tissue were all decreased significantly in DYY groups， while the level of GAS （except for HDYY group） in serum was increased significantly （P＜0.05 or P＜0.01）. The pathology of lung tissue was improved， the percentage of T lymphocytes in LDYY group and the percentages of T lymphocyte and B lymphocyte in MDYY group were all increased significantly （P＜0.05 or P＜0.01）. Compared with the NC group， Bacteroidetes， Actinobacteria， Bacteroides， Alistipes， Bacteroidota， Bacteroides acidfaciens and Alloprevotella were expressed in high abundance， while Firmicutes， Proteobacteria， Lachnospiraceae NK4A136 group， Kineothrix and Clostridiales unclassified were expressed in low abundance. After the intervention of medium-dose DYY， the relative abundance of each microbiota tended to be adjusted back， with different species including Lachnospiraceae_UCG-Ruminococcus， etc. CONCLUSIONS Dayuanyin decoctioncan reduce the lung injury caused by RSV， the mechanism of which may be associated with relieving inflammation， regulating gastrointestinal hormone levels， the percentage of lymphocytes and the abundance of beneficial and harmful bacteria in the intestinal tract.
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ObjectiveTo characterize the chemical constituents of Dayuanyin based on ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry（UPLC-Q-Exactive Orbitrap MS）. MethodThe detection was performed on a Thermo Acclaim™ RSLC 120 C18 column（2.1 mm×100 mm， 2.2 μm）， the mobile phase was acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution （0-7.5 min， 10%-19%A； 7.5-12 min， 19%-22.5%A； 12-23 min， 22.5%-27%A； 23-27 min， 27%-56%A； 27-35 min， 56%-84%A； 35-36 min， 84%-90%A）， the flow rate was 0.3 mL·min-1， and the column temperature was 30 ℃. The data were collected in the positive and negative ion modes by heated electrospray ionization（HESI）， and the detection range was m/z 80-1 200. Combining the retention time of the reference substance， fragment ions， databases such as PubChem and related literature， Xcalibur 3.0 was used to identify the chemical constituents of Dayuanyin. ResultA total of 161 compounds were identified， including 14 alkaloids， 60 flavonoids， 16 terpenoids， 26 saponins， 18 phenylpropanoids， 16 organic acids and 11 others. ConclusionThe established method can effectively and quickly identify the chemical components in Dayuanyin， and clarify its chemical composition， which can provide a basis for the development of compound preparations of this famous classical formula.
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Objective To investigate whether myocardial perfusion impairment and left ventricular (LV) longituadinal systolic dysfunction had happened in patients with coronary slow flow(CSF),and to assess the relationship between them by myocardial contrast echocardiography(MCE) and speckle tracking imaging(STI).Methods A total of 55 patients underwent coronary angiography for angina were enrolled,of those 35 with coronary slow flow phenomenon as CSF group,20 patients with normal coronary angiography as control group.STI and MCE were performed from the apical 4-,3 and 2 chamber views at baseline and after low-dose dobutamine stress echocardiography (LDDSE) in CSF group and control group.STI derived LV global longitudinal strain(GLS) and GLS amplitude of variation(△GLS),and MCE derived myocardial blood flow(MBF),and myocardial flow reserve (MFR) were obtained.And the correlation between GLS/△GLS and MBF/MFR was analyzed.Results At baseline,the GLS and MBF were similar between CSF group and control group(P ＞0.05).After LDDSE,both GLS and MBF were significantly increased in two groups (P ＜0.05).The GLS,△GLS,MBF,and MFR in CSF group were significantly lower than those in control group(P ＜0.05).There was no significant correlation between GLS and MBF at baseline in the CSF group(r =-0.274,P =0.111).However,after LDDSE significant correlation existed between GLS and MBF(r =-0.630,P =0.000).Conclusions LV longituadinal systolic function is impaired in patients with CSF under dobutamine stress test,the impairment of MBF and MFR could be an important contributor to the decrease of LV longituadinal systolic function.
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Objective To study the pericarp surface of Fructus Viticis from three main production area under the scanning electron microscope (SEM), and to provide evidence for distinguishing the Fructus Viticis and its false and the different variety of it. Method Using scanning electron microscope, the pericarp surface characteristics from three main production area were comparative studied. Result The pericarp surface characteristics of the fruits of Vitex trifolia L. var. simplicifolia Cham from Shandong and Jiangxi were extremely similar, but they were obvious different with the fruits of Vitex trifolia L. from Guangxi. Conclusion The pericarp surface characteristics of Fructus Viticis from three main production area under SEM were obvious. The methods was simple and provides the new reference to discriminate the Fructus Viticis from different places.
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Objective To establish a method for the content determination of 3-epibetulinic acid in Nodus Nelumbinis Rhizomatis Charcoal. Methods HPLC was used on Lichrospher C18 column (4.6 mm?150 mm, 5 ?m) with mobile phase of acetonitrile-0.2% glacial acetic acid (75∶25), the flow rate of 1 mL/min, the column temperature of 30 ℃ and the detection wavelength of 207 nm. Results 3-epibetulinic acid has a good linearity within 0.365 4～3.654 0 ?g (r=0.999 6) and the average recovery was 98.83% with RSD=2.47%. The content of 3-epibetulinic acid in Nodus Nelumbinis Rhizomatis Charcoal from different region varied from 0.048 3% to 0.575 8%. Conclusion This method is simple, accurate, with good reproducibility, and can be used for the content determination of 3-epibetulinic acid in Nodus Nelumbinis Rhizomatis Charcoal.
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Objective: To establish a method for the content determination of tannin in Nodus Nelumbinis Rhizomatis. Methods: The content of tannin in Nodus Nelumbinis Rhizomatis was detected by molybdophosphate colorimetric method. Results: The content of tannin in Nodus Nelumbinis Rhizomatis from different drugstores varied from 0.2537% to 1.637%. Conclusion: This method was simple, accurate and can be used for the quality control of Nodus Nelumbinis Rhizomatis.
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Objective:To study the identification method and extract of Nodus Nelumbinis Rhizomatis.Methods:The identification of Nodus Nelumbinis Rhizomatis was carried out by TLC and the extract was determined by 65% alcohol.Results:The relevant spots on TLC plates were clear,and it was feasible to determine the extract of Nodus Nelumbinis Rhizomatis with 65% alcohol.Conclusion:Nodus Nelumbinis Rhizomatis can be identificated by 3-epibetulinic acid and ?-sitosterol as reference substance,there were differents on quantity of the extract of Nodus Nelumbinis Rhizomatis,which bought from different districts.
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OBJECTIVE:To study the identification method of charred Nelumbo nucifera and its extract.METHODS: The identification of charred N.nucifera was carried out by TLC with 3-epibetulinic acid and ?-sitosterol as reference substances.The extract was determined with 65% alcohol as solvent.RESULTS: The TLC spots were clear and determination method was feasible.CONCLUSION:TLC can be applied to determine charred N.nucifera and the contents of 65% alcoholic extracted from charred N.nucifera were different place by place.
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Objective To establish a method for the content determination of c hlorogenic acid in Yinren Granules. Methods The content of chlorogenic acid wa s detected by HPLC. HPLC was performed with Shim- Pack CLC- ODS column(150? 6.0 mm, 5 ? m) at room temperature, 3 % methyl alcohol- methanol( 79 ∶ 2 1) as mobile phase, the detection wavelength at 327 nm, the flow rate being 1. 2 mL/min and sample loop volume being 10 ? L. Results Chlorogenic acid has a good linearity within 0.102～ 1.02? g (r=0.999)and the average recovery was 98. 17 % with RSD being 1.31 % . Conclusion This method is simple, accurate and can be used for the quality control of Yinren Granules.