Immobilized cells on microcarriers for efficient and biomimetic screening of active compounds acting on FGFR4 from Fructus evodiae.

Cell membrane coating strategies have been increasingly researched in new drug discovery from complex herb extracts. However, these systems failed to maintain the functionality of the coated cells because cell membranes, not whole cells were used. Original source cells can be used as a vector for active compound screening in a manner that mimics in vivo processes. In this study, we established a novel approach to fabricate high-density fibroblast growth factor receptor 4 (FGFR4)-HEK293 cells on microcarriers covered with collagen through cell culture and covalent immobilization between proteins. This method enables the efficient screening of active compounds from herbs. Two compounds, evodiamine and limonin, were obtained from Fructus evodiae, which were proven to inhibit the FGFR4 target. Enhanced immobilization effects and negligible damage to FGFR4-HEK293 cells treated with paraformaldehyde were successfully confirmed by immunofluorescence assays and transmission electron microscopy. A column was prepared and used to analyze different compounds. The results showed that the method was selective, specific, and reproducible. Overall, the high density of cells immobilized on microcarriers achieved through cell culture and covalent immobilization represents a promising strategy for affinity screening. This approach highlights the potential of the affinity screening method to identify active compounds from an herbal matrix against designed targets and its prospects for use in drug discovery from herbs.

Integrated metabolomics and network pharmacology analysis to explore pig bile-processed Rhizoma Coptidis and Fructus Evodiae sauce-processed Rhizoma Coptidis in lipopolysaccharide-induced inflammatory response.

Pig bile- and Fructus Evodiae sauce-processed Rhizoma Coptidis (Danhuanglian, DHL; Yuhuanglian, YHL, respectively) are two types of processed Rhizoma Coptidis (Huanglian, HL) in traditional Chinese medicine (TCM). DHL and YHL are representative of HL generated from the subordinate and counter system processing methods, respectively, both noted for their anti-inflammatory effects. How these processing methods can affect the medicinal efficacy of HL remains a hot topic. Here, we discussed the influence of the two methods on the efficacy of final HL products (i.e., DHL and YHL) by comparing their components and anti-inflammatory mechanisms. Enzyme-linked immunosorbent assay was employed to measure inflammatory factors in RAW264.7 cells induced by lipopolysaccharide, and UPLC-Q-Exactive Orbitrap-MS was utilized to analyze the endogenous differential metabolites of RAW264.7 cells treated with HL, YHL, and DHL, and thus to identify the related metabolic pathways. Finally, using network pharmacology, we constructed a "disease-target-differential metabolites-active ingredients" network map. Compared with the control, all three products, HL, YHL, and DHL, significantly reduced IL-6, TNF-α, and IL-1ß levels. 12 differential metabolites related to inflammation were identified and 25 target proteins were overlapping among the three groups. Notably, the anti-inflammatory effects of DHL and YHL were mediated by metabolic pathways such as aminoacyl-tRNA biosynthesis, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, and arginine biosynthesis. Specifically, DHL significantly impacted free fatty acid levels, which was not observed with HL and YHL. On screening, DHL had 9 active ingredients, including three from pig bile, and YHL had 12 active ingredients, with six from the processing excipient Fructus Evodiae. The distinct anti-inflammatory mechanisms and material basis of YHL and DHL were characterized by consistency and distinctiveness. Thus, this study underscores the significant influence of processing methods on the medicinal efficacy of TCMs by revealing their regulatory mechanisms and material bases.

Observation on the therapeutic effect of Wuzhuyu acupoint application on gastrointestinal dysfunction in severe patients / 国际中医中药杂志

Objectives To observe the clinical curative effect of Wuzhuyu acupoint on the treatment of gastrointestinal dysfunction in severe patients. Methods A total of 60 patients with gastrointestinal dysfunction were randomly divided into treatment group and control group, 30 patients in each group. The control group was treated with routine western medicine, and the treatment group was given the acupoint application of Wuzhuyu on the basis of routine treatment. The treatment last 7 days. The clinical efficacy was evaluated by observing bowel sounds recovery time, exhaust time and defecating time, enteral nutrition tolerability, gastrointestinal dysfunction score and acute physiology and chronic health evaluation (APACHE Ⅱ). Results The treatment group showd significantly shorter bowel sound recovery time and anal exhaust time than the control group (t value were 8.811, 5.538, 8.908, all P value <0.01). The enteral tolerance rate was 80.0％ (24/30), and the control group tolerance rate was 53.3％ (16/30). The tolerance rate of treatment group was significantly higher than that of control group (χ2=4.720, P=0.030). The Gastrointestinal dysfunction score and APACHE Ⅱ score in treatment group were significantly lower than control group (t value were 3.873,2.805,all P value <0.01). Conclusions The acupoint application of Wuzhuyu can promote the recovery of gastrointestinal function, increase the tolerance of enteral nutrition, and reduce the severity of the patient's condition and improve the prognosis of patients.

Efficacy observation on prevention of constipation after stroke through abdominal massage and navel therapy with fructus evodiae / 中国实用护理杂志

Objective To study the efficacy on prevention of constipation after stroke through abdominal massage and navel therapy with fructus evodiae.Methods 120 patients of stroke with yin-type syndrome were divided into the observation group and the control group with 60 cases in each group.The control group was given routine nursing,the observation group was given abdominal massage and naval therapy with fructus evodiae based upon routine therapy.Comparisons were made about incidence rate of constipation after stroke and applications of non-retention enema with normal saline.Results There were statistical significances of incidence rate of constipation after stroke and applications of non-retention enema with normal saline.Conclusions Abdominal massage and navel therapy with fructus evodiae are good to constipation after stroke.

Determination of alkaloid from combination of Rhizoma Coptidis and Fructus Evodiae by TLCS / 中成药

AIM: To investigate the changes of alkaloid in Rhizorrm Coptidis and Fructus Evodiae before and after combination. METHODS: The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS : After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochioride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION: In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.

Microcalorimetric studies of drug property differences during combination of Rhizoma Coptidis with Fructus Evodiae after separated decoction / 中草药

Transzuojin Pill.Conclusion Microcalorimetry can be used to investigate the drug property differences during combination of Rhizoma Coptidis with Fructus Evodiae after separated decoction.

Quantitative determination of limonin in traditional Chinese medicine by LC-MS/MS / 中成药

Objective: To develop a simple LC MS/MS method for the measurement of limonin in fructus evodiae, cortex phellodendri, cortex dictamni and its preparation. Methods: Separation was obtained by using a discovery C 18 column (150mm?2.1 mm), The mobile phase:A:water-0.1%HAc;B:acetonitrile-0.1%HAc;The elution program was gradient and the flow rate was 0.2mL?min -1 . The mass detector was set MS1(IS)=187.0 and MS2(Limonin)=471.0 by multiple reaction monitor(MRM) respectively.The sample was added psoralen(IS), then extracted with methanol chloroform(1∶3).Results: Limonin was linear from 25 to 1000ng?mL -1 ( r = 0.9996), The regression equation was C=3.124 As/Ai 1.935?10 -3 . The average recovery was 100.2%.Conclusion: This method is simple, quick, sensitive and exclusive.

Determination of alkaloid from combination of Rhizoma Coptidis and Fructus Evodiae by TLCS / 中成药

AIM:To investigate the changes of alkaloid in Rhizoma Coptidis and Fructus Evodiae before and after combination.METHODS:The content of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride in Rhizoma Coptidis and Fructus Evodiae before and after combination were determined by TLCS.RESULTS:After combination of Rhizoma Coptidis and Fructus Evodiae,contents of berberine hydrochloride,palmatine hydrochloride,jatrorrhizine hydrochloride significantly reduced,among which the reduction of berberine hydrochloride was more obvious.CONCLUSION:In decoction,Fructus Evodiae appears to restrict the alkaloid from Rhizoma Coptidis in order to lower the side effect of Rhizoma Coptidis.