Characterization of chemical constituents of Heibu Yaogao (ointment) / 中国药科大学学报

@#Heibu Yaogao (ointment), which made of Galla chinensis, Scolopendra, etc., is a commonly used hospital preparation with definite clinical efficacy, yet its material basis has not been fully clarified.By ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), the chemical constituents of Heibu Yaogao (ointment) and its crude drug mixture were qualitatively analyzed, and the differences of components before and after preparation were discussed.Scanned by electrospray ionization (ESI) source in positive and negative ion modes, a total of 73 compounds were identified via precise molecular weight, characteristic fragment ions and chromatographic retention time.57 and 65 compounds were detected in crude drug mixture and Heibu Yaogao (ointment) respectively, including tannins, phenolic acids, nucleosides, amino acids, etc.In the process of preparation, the chemical structure changes are mainly concentrated in tannins.The change of components of Heibu Yaogao (ointment) before and after processing were proved by UPLC-Q-TOF-MS/MS, which provides important reference for elucidating the material basis of Heibu Yaogao (ointment), illustrating the rationality of preparation technology and further promoting the quality control of the preparation.

Analysis on Mechanism of Baiyaojian in Treating Ulcerative Colitis Based on Intestinal Flora and Intestinal Mucosal Barrier Function / 中国实验方剂学杂志

Objective:To compare the effects of Baiyaojian before and after fermentation on intestinal flora and expression of Occludin and zonula occludens protein-1 （ZO-1） in intestinal mucosa of mice with ulcerative colitis （UC）， and to explore the mechanism of Baiyaojian and Galla Chinensis in the treatment of UC. Method:Totally 50 mice were randomly divided into 5 groups with 10 mice in each group， one group was randomly selected as blank group， and the other 4 groups were treated with dextran sodium sulfate （DSS） to induce UC model. After modeling， mice in the blank group and model group were given normal saline， and treatment groups were given Mesalazine （0.8 g·kg^-1）， Galla Chinensis decoction （1.8 g·kg^-1） and Baiyaojian decoction （2.7 g·kg^-1） by intragastric administration for 7 days. The 16S rRNA sequencing technology was used to detect the changes of intestinal flora in mouse feces. The histopathological changes of colon tissue were observed by hematoxylin-eosin （HE） staining， and the expression of Occludin and ZO-1 in colon tissue of mice were compared by immunohistochemistry. Result:Compared with the blank group， the abundance and diversity of intestinal flora in UC mice were significantly decreased， and the colonic tissue was thickened with congestion and obvious ulcers， and the expression levels of Occludin and ZO-1 were significantly decreased （<italic>P</italic><0.01）. After treatment with Galla Chinensis and Baiyaojian， the abundance and diversity of flora were improved. At the phylum level， relative abundance of Firmicutes， Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Bacteroidetes decreased significantly （<italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Firmicutes increased and the relative abundance of Bacteroidetes decreased， but there was no significant difference. At the genus level， the relative abundance of <italic>Bacteroides</italic>， <italic>Allobaculum </italic>and <italic>Ruminococcus</italic> decreased significantly （<italic>P</italic><0.01）， the relative abundance of <italic>Roseburia</italic>， <italic>Prevotella</italic>， <italic>Oscillospira</italic> and <italic>Paraprevotella</italic> increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of <italic>Bacteroides</italic> and <italic>Allobaculum</italic> decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the relative abundance of <italic>Prevotella</italic>， <italic>Oscillospira</italic>， <italic>Roseburia</italic> and <italic>Ruminococcus</italic> increased significantly （<italic>P</italic><0.01）. Compared with model group， colon tissue of Galla Chinensis group and Baiyaojian group was recovered obviously， congestion was alleviated， only scattered ulcers were seen. The expression of Occludin and ZO-1 increased， and the expression level of Baiyaojian group was higher than that of Galla Chinensis group. Conclusion:The effect of Baiyaojian is better than Galla Chinensis in the treatment of UC. The mechanism may be through regulating the abundance and diversity of intestinal flora， improving the disorder of intestinal flora and increasing the expression of ZO-1 and Occludin and protecting the intestinal mucosal barrier function for alleviating intestinal inflammation.

Antibacterial effect of different concentrations of Galla Chinensis extract on cariogenic bacteria in a biofilm model / 대한구강보건학회지

OBJECTIVES: Galla chinensis inhibited the adherence of planktonic oral bacteria and acid production by cariogenic bacteria. However, little is known about the relevant conditions of Galla Chinensis extract (GCE) exposure time and concentration and the effect of GCE on the structural and functional activity of cariogenic bacteria. The antibacterial effects of natural G. Chinensis extract on S. mutans , S. sanguinis, and S. oralis biofilms were evaluated in vitro.METHODS: Biofilms formed on glass surfaces were treated with different concentrations of GCE at different exposure times. The effects were assessed by examining the bactericidal activity, acidogenesis, minimum inhibitory concentration, and morphology.RESULTS: There was a statistically significant difference in the bacterial growth inhibition depending on the concentration of the GCE, with bacterial growth being inhibited as the concentration of GCE increased. A concentration of 1.0 mg/ml GCE had similar bactericidal effects against S. mutans and S. oralis biofilms to those produced by 2.0 mg/ml CHX. In the 1.0 mg/ml GCE group, incomplete septa were also observed in the outline of the cell wall, together with disruption of the cell membrane. In addition, there was also a slight exudation of the intracellular content from the bacteria in the 1.0 mg/ml GCE and 2 mg/ml CHX groups.CONCLUSIONS: These results indicate that GCE inhibits the growth of S. mutans, S. sanguinis, and S. oralis with increasing concentrations. It alters the microstructure of S. mutans biofilms. These results suggest that GCE might be a useful anti-bacterial agent for preventing dental caries.

Study on commodity specification and grade standard and quality evaluation of Galla Chinensis / 中国中药杂志

This paper is aimed to study the commodity specification and grade standard of Galla Chinensis,for standardizing market order and guide the market circulation,and provide a basis for standardization of Galla Chinensis. With 33 samples of Galla Chinensis from market as the object of the research,the herbal textual research and market research were carried out. Then the grading indicator were selected by the descriptive statistics,principal component analysis and cluster analysis,combining with production practice,the commodity specification and grade standard of Galla Chinensis were set out. The data of moisture,ash,gallic acid as the internal index were used to analyze the relationship between the quality difference between grades and the appearance characters and the intrinsic composition. Herbal textual research and market research found that the high quality of Galla Chinensis characterized with large,complete,wall thick,grayish brown characteristics,and Galla Chinensis could be divided into gallnuts and horned gall. Through principal component analysis and cluster analysis,combining actual production and herbal record,screening,the length,diameter,single weight,the number of 500 g were determined as 4 grading indicators,the commodity specification was divided into two: gallnuts and horned gall,the grades was divided into two: selected goods and mixed goods. The result of correlation analysis showed there was no significant correlation between the internal index and the appearance characters of Galla Chinensis. The result of multiple comparison showed that the ash content of the selected goods was smaller than that of the mixed goods,but it did not reach significant. The content of gallic acid of the selected goods and the mixed goods of gallnuts were higher than selected goods of horned gall,and higher than mixed goods of horned galls. Using the length,diameter,single weight,the number of 500 g as the appearance characters index. Combining with the herbal textual research and market research,we have divided two specifications and two grades for the commodity specification and grade standard of Galla Chinensis,which can provide a basis for industry standards and national standards.

Transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian based on cold and heat syndrome rats / 中国中药杂志

To reveal the transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian by studying the effect of Galla Chinesis and Baiyaojian on cold and heat syndrome rats. Euthyrox was used to induce the hyperthyrosis model,ice water stimulation was used to induce the cold syndrome model,and different concentrations of Galla Chinesis and Baiyaojian water decoction were administrated by gavage for 15 d continuously. Symptom indexes were evaluated,content of pyruvic acid( PA),ATPase activity in liver and contents of DA,T4,cAMP,5-HT,NE,17-OHCS,TRH and TSH in serum were assayed by enzyme linked immunosorbent assay and spectrophotometry. The rectal temperature,water consumption and body weight of heat syndrome rats in model group were increased,cAMP,NE,17-OHCS,TRH and PA were increased,TSH,Na-K ATPase and Ca-Mg ATPase were increased significantly( P<0. 01),while 5-HT was decreased,compared with those of the blank group( P< 0. 05),the contents of T4,DA,NE,TSH,TRH,cAMP and 17-OHCS were decreased significantly( P<0. 01),PA and Ca-Mg ATPase in WG and BG groups were decreased compared with those of the model group( P<0. 05),and the Galla Chinesis content of WG group was lower than that of BG group,while the contents of 5-HT in WG and BG groups were increased,and the Galla Chinesis content of WG group was higher than that of BG group,with no significant difference of viscera index between heat syndrome rats in blank group,model group and drug groups. The rectal temperature,water consumption and body weight of cold syndrome rats in model group were decreased,DA,T4,cAMP,NE,17-OHCS,TRH,TSH,PA,Na-K ATPase and Ca-Mg ATPase of rats in model group were decreased,whereas 5-HT was increased compared with those of the blank group( P<0. 05),the indexes of heart,lung and kidney were significantly higher than those in the blank group( P<0. 05). Both Galla Chinesis and Baiyaojian can significantly alleviate the symptoms of heat syndrome rats caused by levothyroxine sodium. Galla Chinesis has a better effect than Baiyaojian,but cannot alleviate the symptoms of cold syndrome caused by ice water stimulation,suggestting that the decoction of Galla Chinesis and Baiyaojian are both cold,but Galla Chinesis is colder than Baiyaojian. Cold property in Galla Chinesis fermented Baiyaojian can be relieved. In clinical application,the property of " slight cold" is more accurate than " neutral property" for Baiyaojian.

Galla chinensis extracts and calcium induce remineralization and antibacterial effects of enamel in a Streptococcus mutans biofilm model / 대한구강보건학회지

OBJECTIVES: The purpose of this study was to investigate the combined effects of Galla chinensis extract (GCE) and calcium (CA) on enamel remineralization. The antibacterial effect of G. chinensis on Streptococcus mutans biofilm was also evaluated by examining the bacterial growth, acidogenesis, and morphology of the biofilm in vitro. METHODS: S. mutans biofilm was formed on bovine enamel specimens over a 72-h period and treated for 10 min with 1.0 mol CA, 4,000 ppm aqueous solution of GCE, or a combination of the two (GCE+CA). The enamel specimens were analyzed for enamel surface microhardness after remineralization. We tested the anti-cariogenic effects of GCE based on the inhibition of acid production, antibacterial activity, and morphological changes in S. mutans. The differences between the groups and antibacterial effects were analyzed using one-way analysis of variance. RESULTS: GCE+CA group showed the highest efficacy in enhancing remineralization. The GCE group showed the highest antibacterial activity against S. mutans biofilm. Although the GCE+CA group showed significant antibacterial activity, it was less than that of the GCE group (P < 0.05). Both GCE and GCE+CA groups maintained a pH of approximately 7.0 for 1 h whereas the pH of the control group decreased rapidly from pH 7.3 to pH 6.1. SEM imaging revealed that S. mutans treated with GCE and GCE+CA showed irregular cell wall structure and showed fewer cells in the chain than the typical long chains observed in the control group. CONCLUSIONS: This study found that natural G. chinensis significantly enhances enamel remineralization, and exerts synergistic effects with calcium. It also exerts strong bactericidal activity and inhibits acid production and changes in the microstructure of S. mutans biofilm.

Phenolic constituents of Galla Chinensis and their antiviral activities / 中国药科大学学报

@#Seven compounds in the 95% ethanol extract of Galla Chinensis were isolated by silica gel, ODS, Toyopearl HW-40(s)and RP-HPLC. Their structures were identified on the basis of spectral data and physicochemical properties as benzoic acid, 3, 4-dihydroxy-5-［(3, 4, 5-trihydroxybenzoyl)oxy］-, 5-ethoxycarbonyl-2, 3-dihydroxyphenyl ester(1), trigallic acid(2), 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose(3), shikimic acid(4), gallic acid(5), methyl gallate(6)and ethyl gallate(7). Among them, compound 1 was a novel compound, and compound 2 was firstly isolated from the Galla Chinensis. The inhibition tests of the constituents showed that compound 3 exhibited in vitro antiviral activity against HSV-1 and RSV A strain, RSV Long strain with IC50 values of 13. 3, 3. 3, 3. 3 mmol/L, respectively.

Effect of microcrystalline cellulose on compressibility and moldability of Galla chinensis extract / 中草药

Objective: To study the effect of microcrystalline cellulose (MCC) on compressibility and moldability of Galla chinensis extract (GCE). Methods: Four types of MCC were chosen and mixed with GCE respectively in different proportions. The powder was tabletted by Korsch XP-1 Intelligent Induction Tablet Machine, and the compressibility and moldability were evaluated using plasticity constant, elastic recovery, compression ratio, yield pressure, tensile strength, and their change rules as indexes. Results: The compressibility and moldability of GCE were poor. CeolusKG802 was the best at improving elastic recovery and compression ratio among the four kinds of MCC. AvicelDG improved the plasticity constant and tensile strength most when the adding amount was low, and CeolusKG802 improved them most when the adding amount was high. To yield pressure, CeolusKG802 exhibited greater reduction than the other MCC when added in a low amount. As for a high amount, the result was opposite. Conclusion: MCC has good compressibility and moldability in the compression process; Different types of MCC have different advantages. The change law of compression parameters is obtained by investigating the compression of mixed powders of MCC with different proportions on GCE.

Content Determination and Quality Evaluation of Gallic Acid in Gentiana Rigescens from Different Areas in Guizhou Province / 中国中医药信息杂志

Objective To determine the content of gallic acid and evaluate the quality of Galla chinensis of different habitats in Guizhou Province. Methods HPLC method was used to determine the content of Gallic acid on Alltima C18 column (4.6 mm?250 mm, 5 ?m) with the mobile phase of methanol-water-phosphoric acid (15∶85∶0.085). The flow rate was 1.0 mL/min and the detective wave length was 270 nm. Results The contents of Gallic acid in Galla chinensis of 10 different habitats in Guizhou Province were 51.43%～ 60.25%. Conclusion The quality of Galla Chinensis in different habitats in Guizhou Province are up to the Chinese Pharmacopoeia, and the individual varibility is significant.

Observation of Antibacterial Activity of Galla Chinensis etc. 14 Kinds of Traditional Chinese Medicine against 112 Strains of Staphlococcus Aureus in Vitro / 中国中医药信息杂志

Objective To evaluate the antibacterial activity of Galla chinensis etc.14 kinds of TCM against 112 strains of Staphylococcus aureus in vitro. Method Deterimination of antibacterial activity of Galla chinensis etc.against 112 strains of Staphylococcus aureus was performed by using new agar dilution method. Results The antibacterial efficiency of Galla chinensis, Forsythia suspensa and Acacia catechu (L.) Willd against 112 strains Staphylococcus aureus was good, and the MIC90 was 0.102, 0.244, 1.19 mg/mL respectively. Conclusion Galla chinensis etc. 14 kinds of TCM had strong antibacterial activity agsinst 112 srains of Staphylococcus aureus.

Observation of Antibacterial Activity of Galla Chinensis Ethanol Extractant Against Staphylococcus Epidermidis and Its Morphology Change / 中国中医药信息杂志

Objective To evaluate the antibacterial activity in vitro of Galla Chinensis ethanol extractant(GCEE)against S.epidermidis,and the change of cell morphology of S.epidermidis.Methods Deterimination of antibacterial activity of GCEE against 124 strains S.epidermidis was performed by using agar dilution method,and the changes of cell merphalogy of S.epidermidis was examined by microscope.Results The MIC90 of GCEE against 43 strains of MRSE(methicillin-resistant staphylococcus epidermidis)and 81 strains of MSSE(methicillin-sensitive staphylococcus epidermidis)were 0.288 and 0.144 mg/mL respectively.Under microscope,the cells of S.epidermidis were expansion by degrees,with the edge of cell of S.epidermidis irregular.The substances in cells of S.epidermidis were disappear final.Conclusion Galla Chinensis ethanol extractant had strong antibacterial activity against 124 strains of S.epidermidis,and the cell morphology of S.epidermidis changed.

Effects of Galla Chinensis on the Proliferation of Human Keratinocytes, Melanocytes and Fibroblasts in vitro / 中华皮肤科杂志

Objective To investigate the effects of gallotannic acid, the alcohol extract of Galla chinensis, the alcohol extract of Flos carthami and the water extract of Aloe vera on normal human keratinocytes, melanocytes and fibroblasts in vitro, and get a better understanding of the mechanism of action and the possible adverse effects of Galla chinensis in treating keloid. Methods MTT colorimetric assay was used to detect the effects of gallotannin, colchicine and the later two extracts on the growth activity of the different cells. Results ①The inhibitory action of gallotannin to fibroblasts proliferation was similar to that of the alcoholic extract of Galla chinensis, and the inhibitory action of gallotannin at ≥ 20 ?g/mL to the proliferation of melanocytes was obvious (the proliferation rate was 73.6% - 68% of untreated cells), while gallotannin at ≤ 50 ?g/mL slightly enhanced the growth activity of keratinocytes (the proliferation rate was 111% - 112% of untreated cells). ②Both the alcoholic extract of Flos carthami and the water extract of Aloe vera enhanced the proliferation of keratinocytes in a dose-dependent mode. The water extract of Aloe vera also enhanced the proliferation of fibroblasts. Conclusions The gallotannin may be the main pharmacologically active component of Galla Chinensis for treating keloid, and its inhibitory activity to human melanocytes may implicate its potential adverse effects on the skin.

Chemical constituents of Galla Chinensis / 中草药

Objective To study the chemical constituents of the petroleum and chloroform extracts of Galla Chinensis.Methods Using chromatography on silica gel column and thin-layer chromatography to isolate and purify the compounds;using spectroscopy methods to elucidate their structures;using double dilution to screen antibacterial activities on Fusarium oxysporum.Results Ten compounds were isolated and identified as 2-hydroxy-6-pentadecyl benzoic acid(Ⅰ),4-(pentadec-8-enyl) phenol(Ⅱ),plamitic acid-1,3-dipropyl ester(Ⅲ),?-sitosterol(Ⅳ),pentacosane(Ⅴ),4-hydroxy-3-methoxy-benzoic acid(Ⅵ),plamitic acid(Ⅶ),lauric acid(Ⅷ),myristic acid(Ⅸ),and gallic acid(Ⅹ).Conclusion Compounds Ⅰ-Ⅵ are obtained from Galla Chinensis for the first time.Compound Ⅰ shows moderate inhibitory activity on F.oxysporum.

In- vitro Antibacterial Activity of Ethanol- Extract of Galla Chinensis against Staphycoccus Aureus / 中药新药与临床药理

Objective To evaluate the in- vitro antibacterial activity of e thanol- extract of Galla Chinensis against Staphycoccus aureus (S. aureus).Meth ods The minimum inhibitory concentration (MIC50) of ethanol- extract of Galla Chinensis against 112 strains of S. aureus was detected by using agar dilution method.Results The MIC50 and MIC90 of ethanol- extract of Galla Chinensis aga inst 84 strains of MRSA (methicillin- resistant staphylococcus aureus) were 0. 315, 0.315 mg/mL, and those against 28 strains MSSA (methicillin- sensitive sta phylococcus aureus) were 0.63 and 0.315 mg/mL respectively.Conclusion Ethanol - extract of Galla Chinensis has a strong antibacterial activity against S.aure us.

Extraction of tannins from Galla Chinensis / 中成药

AIM:To optimize the extraction process of tannins from Galla Chinensis. METHODS: Using of HPLC to assay tannins hydrolysis, gallic acid as the content index was analyzed by HPLC, the optimum extraction condition consisted of extraction temperature, the size of material, the solvent extraction times and the extraction time by orthogonal test design. RESULTS: The best condition of extraction process was 10 times the volume of water, extracted three times, 9 hours each time. CONCLUSION: The optimized process is feasible for the extraction of tannins.