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ObjectiveTo investigate the synergistic mechanism of vinegar-processed Olibanum on ulcerative colitis（UC） via the bile acids regulating "gut-liver" crosstalk. MethodRats were randomly divided into normal group， model group， Olibanum group and vinegar-processed Olibanum group. UC model of rats was induced by intracolonic instillation of 2，4，6-trinitrobenzenesulfonic acid（TNBS）. Ultra high performance liquid chromatography-triple quadrupole-mass spectrometry（UPLC-QQQ-MS） was used to perform the qualitative analysis of 30 bile acids in the colon of rats. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect changes in the expression of farnesoid X receptor（FXR）， fibroblast growth factor 15（FGF15） and FGF receptor 4（FGFR4） in "gut-liver" crosstalk at mRNA and protein levels. And with the help of HcoEpiC cell model intervened by conjugated bile acids， simulating the UC state， and according to the different modes of intervention， they were divided into the blank group， conjugated bile acid group， Olibanum group， vinegar-processed Olibanum group and 3-O-acetyl-9，11-dehydro-β-boswellic acid（ADHBA） group. The effect of Olibanum before and after processing with vinegar and the main differential component ADHBA on the mRNA expression of FXR and FGF19 were explored by Real-time PCR. ResultCompared with the normal group， the levels of conjugated bile acids in the model group increased significantly（P<0.01）， and the mRNA and protein expressions of "gut-liver" crosstalk factors FXR， FGF15 and FGFR4 decreased significantly（P<0.05， P<0.01）. Compared with the model group， the content of conjugated bile acids in the Olibanum group and vinegar-processed Olibanum group was significantly decreased（P<0.01）， the mRNA and protein expressions of FXR， FGF15 and FGFR4 were significantly elevated（P<0.05， P<0.01）， and vinegar-processed Olibanum exhibited superior effects than Olibanum. In cellular experiments， a significant decrease in mRNA expression of FXR and FGF19 was observed in the conjugated bile acid group when compared with the blank group（P<0.01）. Compared with the conjugated bile acid group， the mRNA expressions of FXR and FGF19 were significantly higher in the Olibanum， vinegar-processed Olibanum and ADHBA groups（P<0.05， P<0.01）， and the effect of vinegar-processed Olibanum was more favorable. ConclusionVinegar-processed Olibanum may enhance the ameliorating effect on UC by enhancing the down-regulation of conjugated bile acids in the colon and the up-regulation of FXR-FGF15/19-FGFR4 "gut-liver" crosstalk pathway， and ADHBA may be the main material basis for the synergy.
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ObjectiveTo investigate the value of Model for End-Stage Liver Disease (MELD) and Child-Turcotte-Pugh (CTP) score in predicting the prognosis of patients with hepatic sinusoidal obstruction syndrome (HSOS) associated with Gynura segetum (Lour.) Merr. MethodsA total of 49 patients with HSOS associated with Gynura segetum (Lour.) Merr. who were admitted to Beijing YouAn Hospital, Beijing Ditan Hospital, The Fifth Medical Center of Chinese PLA General Hospital, Tianjin Third Central Hospital, and The First Affiliated Hospital of Xinxiang Medical University from January 2012 to July 2018 were enrolled and followed up for three years, with death as the outcome event. MELD and CTP scores were calculated according to the laboratory examination and clinical data on admission, and according to CTP score, the patients were divided into CTP class A (CTP score 5-6) group(n=8), CTP class B (CTP score 7-9) group(n=23), and CTP class C (CTP score ≥10) group(n=18). The patients were divided into death group(n=12) and survival group(n=37) according to the clinical outcome during follow-up. The Mann-Whitney U test was used for comparison of continuous data between groups, and the Kruskal-Wallis H test was used for ranked data. The area under the receiver operator characteristic (ROC) curve (AUC) was used to investigate the ability of CTP and MELD scores in predicting death. The Kaplan-Meier survival curves were used to determine the long-term prognosis of patients with different CTP and MELD scores, and the log-rank test was used for comparison. The ROC curve was used to evaluate the performance of these two scoring systems in predicting death. ResultsA total of 12 patients died during the 3-year follow-up period. The patients with HSOS had a median MELD score of 13.443 (8.792-18.379), and the death group had a significantly higher MELD score than the survival group ［19.84 (15.49-25.41) vs 11.58 (8.60-15.79), Z=-3.511, P＜0.001］. The patients with HSOS had a CTP score of 6-12, and of all 49 patients, 8 (16.3%) had CTP class A HSOS, 23 (46.9%) had CTP class B HSOS, and 18 (36.7%) had CTP class C HSOS. The mortality rate of the patients increased significantly with the increase in CTP score (χ2=16.078, P＜0.05). The mortality rates of the patients with CTP class A, B, and C HSOS were 0.0%, 13.0%, and 50.0%, respectively (χ2=10343, P＜0.05). The Kaplan-Meier analysis showed that the patients with a MELD score of ＜14.294 4 had a significantly better 3-year prognosis than those with a MELD score of ≥14.294 4 (χ2=14.893, P＜0.001). The higher the CTP score, the poorer the 3-year prognosis of patients (χ2=11.083, P＜0.05). CTP class had an AUC of 0.780 (95% confidence interval ［CI］: 0.639-0.922) in predicting the prognosis of HSOS patients, while MELD score had an AUC of 0.840 (95%CI: 0.722-0.958), and there was no significant difference between the two scores (Z=2.63, P＞0.05). ConclusionBoth MELD and CTP scores can predict the risk of death in patients with HSOS, with similar performance in predicting the prognosis of patients, and further studies are needed to validate their clinical value.
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Hepatic sinusoidal obstruction syndrome (HSOS) is a sinusoidal or small venous fibrous occlusive disease characterized by small hepatic vessel damage, especially sinusoidal endothelial cell injury. Exposure to certain exogenous toxins is the main cause of the disease. Based on etiology, HSOS is mainly classified into pyrrolidine alkaloid-related HSOS, hematopoietic stem cell transplantation-related HSOS, and HSOS of unknown etiology. This article summarizes the types of HSOS and reviews the research advances in clinical manifestations, pathogenesis, diagnosis, and treatment of HSOS.
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Objective To investigate whether GLT25D2 gene regulates autophagy in acetaminophen (APAP)-induced hepatotoxicity injury.Methods GLT25D2+/+ wild-type C57BL/6J mice and GLT25D2-/- C57BL/6J mice were selected as subjects. ①In vivo experiment: 20 for wild-type mice and 20 for GLT25D2-/- mice were respectively divided into phosphate buffer (PBS) control group and APAP intervention group according to random number table, with 10 mice in each group. The hepatotoxicity injury model of mice was reproduced by intraperitoneal injection of 25 g/L APAP solution 500 mg/kg. The PBS control group was intraperitoneally injected with the same amount of PBS. The mice were sacrificed immediately after model reproduction, and the liver tissues were harvested. Western Blot was used to detect the expressions of autophagy-related proteins ATG5, ATG7, microtubule-associated protein 1 light chain 3 (LC3) and P62. The ultrastructural changes in liver tissue were observed under electron microscope to observe the level of autophagy. ②In vitro experiment: primary hepatocytes extracted by two-step collagen perfusion from one GLT25D2+/+wild-type mouse and one GLT25D2-/- mouse were divided into two parts respectively. One part was treated with 5 mmol/L APAP solution. The cells were harvested at 0, 8, and 12 hours, and the expressions of autophagy-related proteins ATG5, ATG7, LC3, and P62 were determined by Western Blot. The other part was transfected with the green fluorescent protein-LC3 plasmid (GFP-LC3) for 24 hours. The cells were cultured with PBS (PBS control group) or 5 mmol/L APAP (APAP intervention group) for 12 hours, and the positive expression of GFP-LC3 was observed under the fluorescence microscope, thereby reflecting the expression of autophagosomes.Results ①In vivo experiment: compared with the corresponding PBS control group, the expressions of the positive-associated proteins ATG5, ATG7 and LC3-Ⅱ in liver tissue of the APAPintervention group were down-regulated in the wild-type and GLT25D2-/- mice, while the expression of the negative correlation protein P62 was up-regulated, indicating that the overall level of autophagy decreased after treatment with APAP. Compared with wild-type mice, the expressions of autophagy positive correlation proteins ATG5 and ATG7 were up-regulated in GLT25D2-/- mice (ATG5/β-actin: 1.21±0.29 vs. 0.84±0.19, ATG7/β-actin:1.29±0.14 vs. 1.54±0.40, bothP > 0.05), LC3-Ⅱ expression was slightly down-regulated (LC3-Ⅱ/β-actin: 0.52±0.06 vs. 0.58±0.06,P > 0.05), while negative correlation protein P62 was down-regulated (P62/β-actin: 1.13±0.94 vs. 1.54±0.40,P > 0.05), indicating that the expression of autophagy in GLT25D2-/- mice was higher than that in wild-type mice. Ultrastructural observation under electron microscope showed that the number of autophagosomes in the liver tissue of wild-type mice did not change significantly after APAP intervention as compared with that in PBS control group, but the number of autophagosomes in GLT25D2-/- mice was increased. ②In vitro experiment: with the prolongation of APAP intervention, the expressions of ATG5 and ATG7 in the primary hepatocytes of wild-type and GLT25D2-/-mice were up-regulated, LC3 was slightly fluctuated, and the expression of negative-related protein P62 was gradually down-regulated. The peak value or the trough value reached at 12 hours. It was indicated that the expression of autophagy in APAP-stimulated cells was enhanced with a time-dependent manner. Compared with wild-type mice, the expressions of autophagy correlation proteins ATG5, ATG7, LC3-Ⅱ and P62 were up-regulated in GLT25D2-/- mice at 12 hours (ATG5/β-actin: 0.93±0.09 vs. 0.74±0.06, ATG7/β-actin: 0.80±0.09 vs. 0.65±0.10, LC3-Ⅱ/β-actin:1.35±0.30 vs. 1.15±0.20, P62/β-actin: 0.36±0.02 vs. 0.31±0.03, allP > 0.05), indicating that the expression of autophagy was enhanced after gene knockout. Fluorescence microscopy showed that GFP-LC3 positive cells in both wild-type and GLT25D2-/- mice hepatocytes were significantly increased after APAP intervention as compared with those of PBS control group, and the proportion of GFP-LC3 positive cells in GLT25D2-/- mice was significantly higher than that in wild-type mice (0.64±0.08 vs. 0.36±0.05,P < 0.05).Conclusions GLT25D2 is a negative regulator of autophagy. Knockout of GLT25D2 gene can enhance the autophagy level of APAP-induced hepatotoxicity injury in mice.
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Colistin is a kind of old cationic drug,which can interfere bacterial cell membrane,thus to cause bacterial death.It is mainly used for the treatment of infections caused by Gram-negative bacteria,and its antimicrobial activity against multidrug-resistant Gram-negative bacteria is also very significant.At present,colistin is widely used in veterinary medicine.This article aims to review colistin in chemical,pharmacological,and pharmacokinetic studies,and also summarizes the application and resistance of this drug,which will provide reference for the reasonable selection and use of this drug in animals.
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This paper focused on factors which affected on different color of northern and southern region vinegar-processed frankincense.Meanwhile,contents of six main boswellic acids were also determined to elaborate the influence of heat in chemical components.Vinegar-processed frankincense from northern and southern region was collected.And different temperature and time were used in the processing of frankincense to receive the vinegar-processed frankincense samples.The color difference meter was utilized combining with the PCA statistic analysis method.The Zorbax ExtendC18 chromatographic column (4.6 mm × 50 mm,1.8 μm) was used with acetonitrile-0.1％ phosphoric acid as the mobile phase and gradient elution.The velocity of flow was 1 mL· min-1.The detection wavelength was 210 nm and 250 nm.The column temperature was 30℃.The results showed that the color of northern region processed frankincense was yellow or pale brown.And the southern region processed frankincense was pale brown or dark brown.It showed the difference on processed degree.The L* value of the northern processed frankincense was 75.327 to 80.746 and the L* value of southern processed was 44.321 to 49.527.The a* value of the northern processed frankincense was 5.378 to 6.502 and the a* value of southern processed was 9.423 to 9.978.There was no significant difference on b*.There were certain differences on L* and a* among vinegar-processed frankincense with the same surface color.The color parameter results of self-made vinegar-processed frankincense indicated that along with changes of processing temperature and time,the color,L* and a* change.Even frankincense processed for 30 min with mild fire,it will not achieve the color parameter value of the southern region vinegar-processed frankincense.However,after 11 min processing with medium fire,the color can be achieved.The content determination results showed that four contents,including α-boswellic acid,β-boswellic acid,3-acetyl-α-boswellic acid and 3-acetyl-β-boswellic acid were increased.Contents of 11-carbonyl-3-boswellic acid and 3-acetyl-11-carbonyl-β-boswellic were decreased after being processed.The range of increasing or decreasing by medium fire was higher than mild fire.At the same temperature,as the increasing of processing time,the content has an increasing or decreasing tendency.It was concluded that temperature was the main factor influencing the color of vinegar-processed frankincense from northern and southern regions.Different processing degrees also make influence on the contents of chemical compounds.The color parameter value can be used to evaluate the color of processed frankincense.
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“Multi-component Chinese medicine” (MCC Medicine) is a new TCM concept in recent ten years. It is a new formed TCM product accepted and approved by the new mode (component compatibility of medicines) for TCM research and development, which originated from TCM research and development and TCM pharmaceutics. MCC Medicine contains massive historical accumulation of TCM and distinctive characteristics of the times, which is closely connected with the TCM theory, current trend of the TCM development, clinical treatment requirements, and the development of modern science and technology. In order to promote the development of MCC Medicine, this article reviewed its original background and future trend, with a purpose to make clear the direction for the development of MCC Medicine.
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This study was aimed to analyze the content variations of chemical constituents produced in fresh Radix Rehmanniae (Xian-Di-Huang) after processing into Radix Rehmanniae Recens (Sheng-Di-Huang) and Radix Rehmanniae Preparata (Shu-Di-Huang). HPLC was used in the study of preparing Xian-Di-Huang into Sheng-Di-Huang, and the processing into Shu-Di-Huang. The contents of two newly produced chemical components, which were 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and 5-hydryoxymethyl-furfural (5-HMF). The contents of both chemical components were determined in Sheng-Di-Huang and Shu-Di-Huang bought from the market. The Zorbax SB-C18 column (250 mm í 4.6 mm, 5 μm) was used. The mobile phase was methanol-wa-ter (5:95). The flow rate was 1.0 mL·min-1. The detection wavelength was set at 280 nm. The results showed that no DDMP or 5-HMF was detected in Xian-Di-Huang. However, after processing, DDMP and 5-HMF can be de-tected in both Sheng-Di-Huang and Shu-Di-Huang. The content in Shu-Di-Huang was higher than that in Sheng-Di-Huang. Both contents in Shu-Di-Huang were gradually increased along with processing time. The con-tent reached to the highest level after processing for 24 h and 32 h, respectively. And then, the content decreased. Both DDMP and 5-HMF were detected from three batches of Sheng-Di-Huang and ten batches of Shu-Di-Huang bought from the market. The content of 5-HMF was higher in Shu-Di-Huang than in Sheng-Di-Huang. There was no obvious difference in the content of DDMP between Sheng-Di-Huang and Shu-Di-Huang. It was concluded that DDMP and 5-HMF were produced in the processing Sheng-Di-Huang and Shu-Di-Huang. The contents were gradually increased along with the prolonging of processing time. There was obvious difference in the content of 5-HMF in Shu-Di-Huang and Sheng-Di-Huang.
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Objective To evaluate periphery artery and renal injury in elderly patients with dipper and non-dipper isolated systolic hypertension(ISH). Methods Totally 187 elderly cases were divided into 51 patients with non-dipper hypertension,70 patients with dipper hypertension aged (72.4±5.6) years and 66 cases with normal blood pressure as control according to results of dynamic blood pressure recorder. Ankle-brachial index (ABI),brachial ankle artery pulse wave velocity (baPWV),retinol-binding protein (RBP) and Cystatin C were assessed. ABI and baPWV were determined by a non-invasive automatic waveform analyzer. Results The baPWV value in nondipper group was higher than dipper group [(1869.3±285.6)cm/s vs.(1703.1±235.2)cm/s,q=4.73,P＜0.01],while the value of ABI in non-dipper group was lower than dipper group (1.0 ±0.2vs.1.1±0.2,q=4.74,P＜0.01).The level of Cystatin C was elevated in non-dipper group versus dipper group [(1.4±0.5) mg/L vs. (1.0±0.5)mg/L,q=6.92,P＜0.01]. There were no differences in RBP concentration among the three groups (F=2.39,P＞0.05).At baPWV＞ 1400cm/s,the level of Cystatin C was increased in 47 cases with non-dipper hypertension as compared with 64 cases with dipper hypertension [(1.4±0.5)mg/L vs.(1.1±0.5)mg/L,q=5.59,P＜0.01].Conclusions The elderly patients with non-dipper hypertension may be more easily suffered from periphery artery and renal injury in comparison with dipper hypertension.
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<p><b>OBJECTIVE</b>To establish method for determining the contents of alpha-pinene and octyl acetate in Boswellia serrata, in order to provide preference for making quality standards for B. serrata and processed B. serrata.</p><p><b>METHOD</b>Application of orthogonal design was employed to optimize the solvent, solvent quantity and extraction time. The GC-MS analysis was performed on a Rxi-5ms silica capillary column, running in the electron impact (EI) mode, with ion trap and injector temperature of 200 degrees C and 250 degrees C, respectively. The column oven was initially 50 degrees C and was held for 1 min after injection, followed by temperature ramping at 5 degrees C x min(-1) up to 130 degrees C, holding for 1 min. 1 microL of samples solution were injected in the split mode (1:60). Helium was the carrier gas. The mass spectrometer was set to scan m/z 45450 with an ionizing voltage at 70 eV.</p><p><b>RESULT</b>Sample solutions were prepared for 50-fold dose by ultrasonic extraction with hexane for 30 min. The content of alpha-pinene and octyl acetate in 10 batches of B. serrata were 0.021 3-0.149 5, 2.519 6-9.098 0 mg x g(-1), respectively. And, those of alpha-pinene and octyl acetate in processed B. serrata were 0.015 9-0.065 9, 0.801 0-12.812 2 mg x g(-1).</p><p><b>CONCLUSION</b>The method is a stable and reliable for determining the contents of alpha-pinene and octyl acetate in B. serrata.</p>
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Acetates , Boswellia , Chemistry , Gas Chromatography-Mass Spectrometry , Methods , MonoterpenesABSTRACT
Objective To optimize the extraction method of salviae miltiorrhizae madix et rhizoma in Huoluo-Xiaoling granules. Methods According to the contents of Salvianolic acid B and Tanshinone Ⅱ A, the extraction method was established by comparing different solvents (water and 70% ethanol)and extracting modes (compound extraction and single herb extraction). Then orthogonal design was used to determine the optimum extraction method. Results Considering the contents of salvianolic acid B and tanshinone Ⅱ A, 70% ethanol extract was better than water extract and compound extraction was better than single herb extraction.The optimum extraction condition was 70% ethanol in eight times of the herbs weight, extracted for 1h by 3times. Conclusion The extraction method was simple and stable.
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<p><b>OBJECTIVE</b>To develop an HPLC method for determinating the contents of five boswellic acids in Boswellia serrata.</p><p><b>METHOD</b>Analysis was performed on a zorbax SB C18 column eluted with acetonitrile-0.1% phosphoric acid in water as mobile phases in gradient elution and the detection wavelengths were 210 nm and 250 nm.</p><p><b>RESULT</b>The five ingredients were separated well. The content ranges of alpha-boswellic acid, beta-boswellic acid, 3-acetyl-beta-boswellic acid, 11-keto-beta-boswellic acid and 11-keto-beta-acetyl- boswellic acid were 8.68-16.1, 53.5-246.9, 38.4-192.9, 4.48-5.81, 32.7-44.2 mg x g(-1), respectively.</p><p><b>CONCLUSION</b>The contents of five individual boswellic acids were different in 12 batches of B. serrata samples.</p>
Subject(s)
Boswellia , Chemistry , Chromatography, High Pressure Liquid , Plant Extracts , TriterpenesABSTRACT
<p><b>OBJECTIVE</b>To study the relation between the content of 5-hydroxymethyl-furfural (5-HMF) and the degree of floating sugar in root of Achyranthes bidentata.</p><p><b>METHOD</b>An HPLC method was applied with a Waters Symmetry C18 3.9 mm x 150 mm (5 microm) column by using methanol-water (12:88) as the mobile phase at a flow rate of 1.0 mL x min(-1) and a UV detection of 280 nm.</p><p><b>RESULT</b>Along with the degree's deepening of floating sugar, the content of 5-HMF varied with the different shades of the sample. The content was 10 times higher in the black sample (highest degree of flowing suger) than that in the yellowish sample (normal). The concentrations of 5-HMT in five yellowish samples of roots of A. bidentata were 0.162 mg x g(-1) to 0.332 mg x g(-1).</p><p><b>CONCLUSION</b>The content increasing of 5-HMF in the root of A. bidentata was related to the degree of flowing sugar.</p>
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Achyranthes , Chemistry , Carbohydrates , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Furaldehyde , Plant Roots , ChemistryABSTRACT
Objective To inspect the quality of Fructus Aurantii Immaturus formula granule. Methods Choose three batch Fructus Aurantii Immaturus to prepare formula granule and decoction. Zobax Extend C18 column was used with acetonitrile-water-phosphoric acid-sodium lauryl sulphate (12∶87∶1∶0.2) as the mobile phase, the detection wavelength was 275 nm. Result There were no differences in elution peak quantity and peak position in the HPLC spectrum of Fructus Aurantii Immaturus formula granule with corresponding cut crude drug and decoction. The relative peak area in the corresponding peak of formula granule had no significant difference with decoction, but had certain differences with cut crude drug. Conclusion HPLC spectrum showed that Fructus Aurantii Immaturus formula granule and decoction had high similarity.
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Objective To select the best condition of preparation process for the water-extraction and alcohol-precipitation method of Radix astragali in “Leifengguan” granules. Methods The effective compound astragaloside IV of Radix astragali was determined with the methods of TLC-scanning. The preparation process was screened with orthogonal design [L9(34)] and single factor analysis. Results The best condition of preparation process for water-extraction of Radix astragali was A3B2C2 that the drug materials were decocted for 3 times with 8 times amount of water, each time for 1 hour. After the solution was concentrated to proportion as 1 g/mL (drug materials/solution), alcohol was added to the solution to 60% alcohol. Conclusions The optimized preparation process was found to be stable with a good reproducibility.
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Objective To study the method for distinguishing the crude and processed Radix of Polygonum multiflorum Thunb. by TLC. Methods Some of the samples were collected from different cities in China, some of them were prepared following the pharmacopoeia standard or local standard. TLC conditions: 5-hydroxymethyl-furfural (5-HMF) was used as a standard matter with the developing solvent of petroleum ether (60～90 ℃)-ethyl acetate (1∶1) on a silica G thin layer plate, sprayed with 15% ?-naphthol in ethanol solvent, heated and detected in daylight. Results TLC chromatogram showed that crude and processed product of Polygonum multiflorum Thunb. were different. Conclusion TLC method for distinguishing the crude and processed Polygonum multiflorum Thunb. was established.
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Objective: To determinate the content and stability of 2,3,5,4′ stilbene glucoside in prepared Radix Polygoni Multiflori from differet areas.Methods:The content of 2,3,5,4′ stilbene glucoside was determined by HPLC, and the test of its stability was carried out on high temperature and 60 Co ? ray radiation. Results: The contents of 2,3,5,4′ stilbene glucoside in prepared Radix Polygoni Multiflori from different areas varied from 0.127% to 4.150%. 2,3,5,4′ stilbene glucoside was unstable under high temperature especially in solution, but it was stable under 60 Co ? ray radiation. Conclusion: The qualities of prepared Radix Polygoni Multiflori are different in different areas.
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OBJECTIVE: To compare the contents of main chemical components in prescription granules and decoction of Fructus aurantii immaturus. METHODS: 3 batches of cut crude drugs of Fructus aurantii immaturus were selected to prepare the granules and decoction. The assaying of water soluble extractive was conducted according to the specification of China Pharmacopeia. The assaying of hesperidin and synephrine was conducted by HPLC. RESULTS: There was no obvious difference between water soluble extractive of granules and that of decoction, while the contents of hesperidin and synephrine were higher in the prescription granules than in the decoction. CONCLUSIONS: The results provide supporting basis for the labeled amount of prescription granule of Fructus aurantii immaturus.