Exploring Synergistic Mechanism of Processing with Vinegar on Olibanum in Treatment of Ulcerative Colitis Based on Intestinal Flora / 中国实验方剂学杂志

ObjectiveTo explore the improving effects and its synergistic mechanism of Olibanum before and after processing with vinegar on glycodesoxycholic acid（GDCA） intervention in mice with ulcerative colitis（UC） based on the perspective of intestinal flora. MethodC57BL/6J male mice were randomly divided into the normal group， model group， GDCA group， Olibanum group（1.5 g·kg-1） and vinegar-processed Olibanum（1.5 g·kg-1） group， with 6 mice in each group. Mice in the normal group drank water freely， and mice in the other groups were given 2% dextran sulfate sodium（DSS） periodically to establish a UC mouse model. During the modeling， GDCA group， Olibanum group and vinegar-processed Olibanum group were intervened by intraperitoneally injection of GDCA（0.05 g·kg-1）. From the 13th day after modeling， Olibanum group and vinegar-processed Olibanum group were given the corresponding doses of drugs by gavage， once a day， for 36 d. During this period， the body mass of mice was recorded and the disease activity index（DAI） was assessed. On day 48， faeces were collected for 16S rRNA and metagenomic sequencing to analyse changes in intestinal flora. On the 49th day， hematoxylin-eosion（HE） staining was used to observe the colon histological lesions， enzyme-linked immunosorbent assay（ELISA） was used to determine serum levels of tumour necrosis factor-α（TNF-α）， interleukin（IL）-1β and IL-6， and Spearman correlation analysis was used to explore the correlation between differential bacterial species and inflammatory factor levels. ResultCompared with the normal group， the model group showed a significant decrease in body weight（P<0.01）， a significant increase in DAI（P<0.05）， and a significant increase in TNF-α， IL-1β and IL-6 levels（P<0.01）， and there was partial infiltration of inflammatory cells in the colon. Compared with the model group， mice in the GDCA group showed a significant decrease in body weight， a significant increase in DAI and levels of TNF-α， IL-1β and IL-6（P<0.01）， and severe disruption of colonic crypt structure， extensive infiltration of inflammatory cells， and a significant decrease in goblet cells. Compared with the GDCA group， both the Olibanum and vinegar-processed Olibanum groups showed a significant recovery in body weight， a significant decrease in DAI and levels of TNF-α， IL-6 and IL-1β（P<0.05， P<0.01）， and the modulating effect of vinegar-processed Olibanum was significantly better than that of Olibanum. Alpha diversity showed that Chao1 index of UC mice significantly increased（P<0.01） and Shannon index decreased significantly（P<0.05） in UC mice after GDCA intervention. Beta diversity showed that the microbial community structure of the 5 groups had significant changes， Olibanum and vinegar-processed Olibanum could modulate the changes in the structure of the intestinal flora in UC mice after GDCA intervention. Microbial sequencing results indicated that， compared with the normal group， the Firmicutes/Bacteroidetes ratio in the model group was significantly higher（P<0.05）， and the relative abundance of 3 genera and 5 species of flora changed significantly（P<0.05， P<0.01）. Compared with the model group， the Firmicutes/Bacteroidetes ratio in the GDCA group was significantly higher（P<0.05）， the relative abundance of 7 pathogenic bacterial genera and four species was significantly increased（P<0.05， P<0.01）， and the relative abundance of three beneficial bacterial genera and Bacteroides_intestinalis was significantly decreased（P<0.05， P<0.01）. Olibanum group and vinegar-processed Olibanum group could modulate the Firmicutes/Bacteroidetes ratio， the relative abundance of pathogenic bacteria and beneficial bacteria， and the vinegar-processed Olibanum group was significantly superior to Olibanum group in terms of modulating the Firmicutes/Bacteroidetes ratio， the relative abundance of the three genera and five species of bacteria（P<0.01， P<0.05）. Correlation analysis showed that the relative abundance of Bacteroides_intestinalis was negatively correlated with the levels of TNF-α， IL-6 and IL-1β， the relative abundance of Prevotella_sp_CAG873， Bacteroides_sp_CAG927， Bacteroidales_bacterium_52_46 and Bacteroidales_bacterium was positively correlated with TNF-α， IL-6 and IL-1β levels. ConclusionGDCA can exacerbate UC colonic inflammation， and Olibanum and vinegar-processed Olibanum have an ameliorative effect on GDCA-mediated UC， with the vinegar-processed Olibanum showing a stronger ameliorative effect， the mechanism may be related to the regulation the abundance and structure of intestinal beneficial and pathogenic bacteria， and the reduction of inflammatory factor levels.

Exploring Synergistic Mechanism of Vinegar-processed Olibanum in Treatment of Ulcerative Colitis Based on Bile Acid-regulated "Gut-liver" Crosstalk / 中国实验方剂学杂志

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.

Effects of Processed Temperature And Time on Color and Contents of Six Types of Boswellic Acids in Frankinense / 世界科学技术-中医药现代化

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.

Review on Original Background of Multi-component Chinese Medicine and Its Future Expectation / 中国中医药信息杂志

“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.

Content Changes of Chemical Constituents Produced in Fresh Radix Rehmanniae after Processing / 世界科学技术-中医药现代化

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.

Determination of alpha-pinene and octyl acetate contents in Boswellia serrata / 中国中药杂志

<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>

Determination of five boswellic acids in Boswellia serrata / 中国中药杂志

<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>

Study on the extracting method of salviae miltiorrhizae madix et rhizoma in Huoluo-Xiaoling granules / 国际中医中药杂志

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.

Determination of 5-hydroxymethyl-furfural in root of Achyranthes bidentata expressing different degree of floating sugar / 中国中药杂志

<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>

Contrasting Analysis on HPLC Spectrum of Fructus Aurantii Immaturus Formula Granule with Corresponding Cut Crude Drug and Decoction / 中国中医药信息杂志

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.

TLC for Distinguishing the Crude and Processed Radix of Polygonum Multiflorum Thunb / 中国中医药信息杂志

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.

Determination of content and stability of 2,3,5,4′-stilbene glucoside in prepared Radix Polygoni Multiflori from different areas / 中成药

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.

Comparison of Main Chemical Components in Prescription Granules and Decoction of Fructus Aurantii Immaturus / 中国药房

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.