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We explored the mechanism of patchouli oil in the treatment of inflammatory bowel disease (IBD) based on network pharmacology and differentially expressed genes in macrophages. The chemical composition of patchouli oil was detected by GC-MS, targets for active components were collected through TCMSP and Swiss Target Prediction platform, and targets for treatment of IBD were retrieved from DrugBank, GeneCards, OMIM, PharmGkb, and TTD databases. The intersection targets were merged, Cytoscape software was used to construct the "component-to-intersection target" network, and protein-protein interaction (PPI) network was drawn with String platform. The intersection targets were enriched for GO and KEGG enrichment analysis on Metascape platform, and the molecular docking of AutoDock Vina was used to verify the analysis results. The macrophage chip data was downloaded, and the differential genes were obtained by using R software. KEGG signaling pathway analysis of differentially expressed genes were performed by DAVID platform. Real-time fluorescence quantitative PCR was used to verify the screened components in the cell model in vitro. The 14 main components of patchouli oil corresponded to 112 targets, and the intersection obtained 97 common targets of patchouli oil for IBD treatment. GO enrichment analysis yielded 53 items. Eighteen items were obtained by KEGG enrichment analysis, involving cAMP signaling pathway, Notch signaling pathway, adhesion connection, Th17 cell differentiation and other signaling pathways. Molecular docking showed that the selected active components of patchouli oil had good binding activity with the targets. Differentially expressed genes were enriched in inflammatory pathways such as Toll-like receptors, JAK-STAT and NF-κB signaling pathways. q-PCR showed that patchouli oil, patchouli alcohol, pogostone can reduce the mRNA levels of cytokines (TNF-α, IL-1β, IL-6, and IL-23) and up-regulate the mRNA levels of tight junction proteins (occludin and claudin-1) in the inflammatory model of NCM460 normal colon epithelial cells. Patchouli alcohol can significantly reduce the levels of TNF-α, IL-6, and IL-1β inflammatory factors in RAW264.7 macrophages induced by LPS. This study revealed the multi-component, multi-target and multi-pathway of patchouli oil, and confirms the anti-inflammatory effect of patchouli oil and its main components in the inflammatory cell model in vitro and the protection of intestinal epithelial barrier integrity function, which provides a theoretical basis for further elucidating the mechanism of patchouli oil in the treatment of IBD.
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Objective: To explore the influence of ancient processing method black bean "nine cycles of steaming and drying" and modern pharmacopoeia processing method continuous steaming with black bean decoction on the main components of Polygoni Multiflori Radix (PMR). Methods: Simulating the time arrangement of "nine cycles of steaming and drying", samples were prepared using three processing methods: ancient method that raw PMR (rPMR) and black bean were steamed in layers and then dried, modern method that rPMR were steamed with black bean decoction and then dried, the method recorded in Chinese Pharmacopoeia that rPMR were steamed continuously with black bean decoction; The determination method of 12 components in rPMR and processed PMR (pPMR) was established using ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UPLC-QqQ-MS/ MS), and 12 components in all samples processed by different methods were determined; The results was analyzed combining with t test, principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). Results: A reliable UPLC-QqQ-MS/MS method was established for the determination of emodin, physcion, rhein, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, cis-2,3,5,4'-tetrahydroxylstilbene-2-O-β-D-glucoside (cis-THSG), trans-2, 3,5,4'-tetrahydroxylstilbene- 2-O-β-D-glucoside (trans-THSG), polydatin, resveratrol, epicatechin, rutin and hyperoside. With the prolongation of steaming time, the content of 12 effective components changed obviously: the content of free anthraquinone was decreased first and then increased; The content of anthraquinone glycoside, cis-THSG, polydatin and hyperoside was increased first and then decreased; The content of trans-THSG, resveratrol, epicatechin and rutin was decreased; The components content were closely related to the auxiliary materials, steaming operation methods and processing time, the influence of operation methods was greater than that of auxiliary materials on the quality of pPMR. Conclusion: The ancient processing method steaming with black bean and drying could not be equated with the modern pharmacopoeia processing method continuous steaming with black bean decoction in terms of the content of effective components. The result provides experimental basis for inheriting and developing the traditional processing method of PMR.
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BACKGROUND:Bisphosphonates-related osteonecrosis of the jaw (BRONJ) is thought to be caused by trauma to dentoalveolar structures after dental extraction and implantation that have a limited capacity for bone healing followed by infection,suppuration and pain,further inducing equestration,but the pathogenesis is poorly understood.OBJECTIVE:To establish a stable animal model of BRONJ,and to observe its clinical manifestations,radiological changes and pathological characters,thereby paving ways for further understanding the pathogenesis of BRONJ.METHODS:Twenty-four female Sprague-Dawely rats were randomly allocated into experimental and control groups (n=12 per group).The rats in the experimental group were given the intraperitoneal injection of 0.2 mg/kg zoledronic acid,and the controls received the intraperitoneal injection of same volume of normal saline,three times weekly for consecutive 12 weeks.Subsequently,left first mandibular molars were extracted under general anesthesia,and the bone healing was observed.At 8 weeks after tooth extraction,all the rats were euthanized.The rat left jaws were examined by X-ray and micro-CT to explore the imaging changes.The histopathological characteristics of the mandibular soft and hard tissues were investigated by hematoxylin-eosin staining and Masson staining,to further testify the BRONJ model.RESULTS AND CONCLUSION:At 8 weeks after tooth extraction,the wound was unable to heal,and nonvital bone tissues were visible in the experimental group.The findings revealed on radiographies and pathological sections were in accordance with the characteristics of BRONJ.Masson staining results showed slender collagen fibrils around the dental extraction wound were slender in the experimental group.These results indicate that the intraperitoneal injection of zoledronate combined with tooth extraction can induce BRONJ in rats.The soft tissue toxicity and anti-angiogenesis of zoledronic acid can promote the occurrence of BRONJ,and the local acidic microenvironment caused by trauma probably is involved in the occurrence of BRONJ.
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<p><b>OBJECTIVE</b>To observe the therapeutic effect and safety of perineal, crissal and progenital pruritus treated with acupuncture according to differentiation.</p><p><b>METHODS</b>Self-control method was applied in these 32 cases. Changqiang (GV 1), Huiyin (CV 1), Qugu (CV 2), Sanyinjiao (SP 6) and Ashi points etc. were punctured as main points, and adjunct points were added according to differentiation: Taichong (LR 3) and Ququan (LR 8) etc. were added for wind and heat excess of liver meridian, Xuehai (SP 10) and Quchi (LI 11) etc. were added for blood deficiency and wind dryness. Itchiness, skin lesions sign scores and therapeutic effects were observed before and after treatment.</p><p><b>RESULTS</b>The total scores of itchiness before and after treatment were 6.06 +/- 1.46 and 2.19 +/- 1.71 respectively, and the total scores of skin lesions sign were 4.38 +/- 2.21 and 1.50 +/- 1.44, indicating that the scores and the total scores of itchiness and skin lesions sign reduced obviously after treatment (P < 0.05, P < 0.01); the cured and markedly effective rate was 73.4% (11/15) for wind and heat excess of liver meridian, and 70.6% (12/17) for blood deficiency and wind dryness, presenting similar therapeutic effect (P > 0.05). Hematoma or ecchymosis appeared in 2 cases, and disappeared spontaneously after 2-3 days, without obvious adverse reaction.</p><p><b>CONCLUSION</b>Simple perineal, crissal and progenital pruritus treated with acupuncture according to differentiation is effective, safe and applicable.</p>