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ObjectiveTo investigate the mechanism of Gegen Qinliantang（GQT） on the intestinal flora of antibiotic-associated diarrhea（AAD） by 16S rRNA sequencing and network pharmacology. MethodSixty SD rats were randomly divided into six groups（n=10）， including blank group， model group， GQT high-， medium- and low-dose groups（10.08， 5.04， 2.52 g·kg-1） as well as Lizhu Changle group（0.15 g·kg-1）， except for the blank group， each group was given clindamycin（250 mg·kg-1） by gavage once a day for 7 consecutive days. After successful modeling， the blank group and the model group were given equal volumes of normal saline by gavage. The other groups were given corresponding doses of drugs by gavage for 14 days. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） was used to screen the active components and targets of GQT， GeneCards， Online Mendelian Inheritance in Man（OMIM） database， Pharmacogenetics and Pharmacogenomics Knowledge Base（PharmGKB）， DrugBank and DisGeNET were used to search for AAD disease targets. The drug-disease common targets were obtained by R software. STRING was applied to analyze the target protein-protein interaction， and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis was performed. Then hematoxylin-eosin（HE） staining was used to observe the pathological changes of the colon， and 16S rRNA sequencing of AAD colon content flora structure further verified the results of network pharmacology. ResultThrough network pharmacology， it was found that 238 active components were screened from GQT and acted on 276 component targets， among which quercetin， puerarin， wogonin and apigenin were the main core components of GQT， 1 097 AAD disease targets and 127 drug-disease intersection targets. The protein-protein interaction network mainly included core targets such as protein kinase B1（Akt1）， interleukin（IL）-6 and IL-1β， which were mainly enriched in the IL-17 signaling pathway. It was verified through animal experiments that compared with the blank group， the colon structure of the model group was seriously abnormal， the intestinal epithelial columnar cells were damaged， the goblet cells were reduced， and a large number of inflammatory cells were infiltrated. Compared with the model group， the colon structure of the GQT high-dose group improved， but there were still abnormalities， the colon structure of GQT medium- and low- dose groups and Lizhu Changle group improved significantly and reached the normal level. GQT could improve the structural diversity of AAD intestinal flora. At the phylum level， the abundance of Firmicutes was increased and the abundance of Bacteroidetes was decreased. At the genus level， the abundance of Lactobacillus was increased， and the abundances of Prevotella and Bacteroides were decreased. Among them， Lactococcus could be used as a biomarker for AAD treatment with GQT， and the prediction of functional metabolism of intestinal flora revealed that GQT could promote acetate and lactate metabolic pathways in the intestine. ConclusionGQT may activate IL-17 signaling pathway by acting on the targets of Akt1 and IL-6 through key components such as quercetin and wogonin， and improve the abundance of Lactococcus in the intestinal tract as well as acetate and lactate metabolic pathways， so as to play a role in repairing the intestinal barrier for the treatment of AAD.
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Objective:To investigate the effects of Huang-Lian-Jie-Du-Decotion containing serum for the autophagy of RAM264.7 macrophages ,and to explore its possible anti-atherosclerosis mechanism .Methods:The SD rats were intervened with large , middle and small dose Huang-Lian-Jie-Du-Decotion to prepare containing drug serum .Cell proliferation was tested by MTT assay .The gene and protein expressions of Beclin 1, mTOR was assessed by fluorescent quantitative PCR and Western blot method .Results:Compared with the normal serum group ,Huang-Lian-Jie-Du-Decotion containing serum enhanced expression of the mRNA and protein level of Beclin1,and decreased the expression of mTOR and p-mTOR(P<0.05 or P<0.01).Conclusion:Huang-Lian-Jie-Du-Decotion containing serum could regulate the expressions of Beclin 1 and mTOR.These might be one of the mechanisms for preventing atheroscle-rosis.
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Objective To study the effect of Danggui Buxue Decoction on red cell immune function in mice with morphine abstinence syndrome. Methods 50 mice were randomly divided into normal group,positive control group, high, medium, low dose group,Hindlimb injection of morphine in morphine dependent mice model was established.Erythrocyte membrane were measured in five groups of mice micro viscosity, erythrocyte deformation index; Determination of red cell immune complex rosette rate using the method (CIC), red cell C3b receptor rosette rate;and to observe the five group of regulator factor enhance rate (RFER) and immune adherence inhibitor factor (RFIR) levels.Results Erythrocyte membrane microviscosity in model group were significantly higher than those in normal group, medium group and low group (P<0.05),red cell deformability index was lower than that of normal group, drug medium group and drug group low (P<0.05).Drug high group and medium group,immune complex rosette rate lower than in model group(P<0.05), C3b receptor rosette rate was significantly higher than that in the model group (P<0.05).Conclusion Low dose group and high dose group Danggui Buxue Decoction raises the red cell immune function in morphine withdrawal mice, improves the withdrawal symptoms, rapids rehabilitation of the body, has good clinical value.
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Objective:To investigate effects of viola on macrophage TOLL-like receptor expression ,and tentatively explore the partial mechanism of viola intervention on macrophage function.Methods: Using viola water decoction lavage intervention in clean grade SD rats of conventional preparation containing serum ,In a certain concentration coculture with mouse peritoneal macrophages in 96-well plates.After 24 hours,cell phagocytosis activity was determined by neutral red uptake assay ,changes in expression levels of TLR-1,TLR-2,TLR-3,TLR-4,TLR-5 mRNAs were determined by RT-PCR.Results:Compared with the same concentration of normal serum group :(1 ) viola-containing serum group macrophage activity was significantly increased ( P0.05 ) ,and the expression level of TLR-5 was significantly increased in each group ( P<0.01 ).Conclusion:Viola-containing serum has obvious promoting effect on macrophage phag-ocytosis,the mechanism of which may be related to that this drug regulates and controls part of macrophage TOLL -like receptor expres-sion.
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Objective: To study the effect of ecological factors on contents of gastrodine, preliminary discuss the relations between the soil ecological factor and the quality of gastrodine. Methods: To collected wild and cultivated gastrodia rhizome and growth soil sample from the different habitat in different harvesttime. To examine microorganism quantity, the contents of soil organic matter, the contents of soil metallic, the soil pH value and the contents of gastrodine in the soil, and analyze the relations between the soil ecological factor and the quality. Results: In collection samples, it’s showed negative correlation (P0.05). Conclusion: The bacteria/fungus constitution condition, the contents of soil organic matter and the soil pH value were important ecological factor that affect the gastrodia quality.
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Objective: To observe the effect of polysaccharide ingredients from six TCM complex prescriptions on releasing the cytokine(CK) level of mouse celiac macrophages (M?), to explore the possible mechanism and direct the extractionand separation of active components for above compounds. Methods: The total polysaccharide ingredients of six complex prescriptions were prepared: Siwu Decoction, Sijunzi Decoction, Liuweidihuang Decoction, Guizhi Decoction, Longdanxiegan Decoction and Yupingfeng Pulveres, then the mouse celiac M? were incubated together in 96 holes board. Furthermore, levels for the latter to release CK, including IL-1?, IL-6, IL-8 and TNF-?, were measured. Results: Polysaccharide ingredients of each complex prescription could obviously accelerate mouse celiac M? to release one or more CK, and the e ects had concentration otherness (P
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AIM:To establish macrophage mannose receiver Model(MMR),and use it to screen active component with mannose receiver(MR)as target from traditional Chinese herbs.METHODS:The mouse abdominal macrophages was hatched with D-mannose and D-galactose of the different concentration,and the flow cytometry and fluorescence microscope were used to measure the antagonistic effect of M-FITC-BSA(Mannose-fluorescein isothiocyanate-bovine serum albumin)with D-mannose and D-galactose.After the MMR screening model was established to screen MR union ingredients of polysaccharide ingredients from six compound prescriptions,such as Siwu Decoction and so on.RESULTS:Both of measuring methods showed that when D-mannose concentration increased the relevance ratio of M? marked with M-FITC-BSA decreases(P