Objective:To analyze any effect of combining hyperbaric oxygen with qingkailing on the metabolic biomarkers of consciousness disorder (CD) caused by brain trauma using metabolomics technology. Also, to screen out diagnostic markers for clinical treatment and prognosis.Methods:Thirty-six patients with a CD caused by brain trauma formed the observation group, while 40 healthy gender-, age- and ethnicity-matched individuals were the control group. Both groups were given routine supportive treatment, while the observation group additionally received hyperbaric oxygen and oral qingkailing medication. Before and after 14 days consciousness disturbance was evaluated using the Glasgow Coma Scale (GCS). Venous blood was collected for metabolomic analysis using liquid chromatography linked to a mass spectrometer to screen out specific metabolic biomarkers. Metabolic markers associated with the disease and of diagnostic significance were thus identified.Results:After the treatment the average GCS score of the observation group had improved significantly and the degree of coma was significantly relieved. Twenty-one metabolic markers were found to be significant, with creatine, proline, uric acid, acetyl-L-carnitine, histidine, proline leucine, tryptophan and 9E-octagenoic acid potentially of high diagnostic value. After the treatment, all of those markers came close to the levels observed in the healthy control group.Conclusions:Proline, leucine, 9E-octagenoleic acid, uric acid and acetyl-L-carnitine could be used for diagnosis and evaluating efficacy with such patients. Hyperbaric oxygen supplemented with qingkailing can relieve coma, correct metabolic disorders and accelerate patients′ awakening. Metabolomics provides a new method for identifying endogenous metabolic abnormalities in patients with post-traumatic consciousness disorders. It can be useful in prognosis and clinical treatment.

Objective This study aimed to systematically investigate the pharmacological substances and potential mechanisms of Zhenzhu Tongluo Wan in the treatment of peripheral nerve disorders.Methods Based on the TCMSP database and related literature,the active ingredients of Zhenzhu Tongluo Wan were determined.Target proteins for these active ingredients and those associated with peripheral neuropathy were predicted using the TCMSP,SwissTarget Prediction,GeneCard,OMIM,and DisGeNET databases.The intersection of these two sets of proteins was once considered a key target for the treatment of peripheral neuropathy.A protein-protein interaction(PPI)network of the intersection targets was constructed using the STRING database and analyzed using Cytoscape 3.9.1 software.Key targets obtained from the network were subjected to GO enrichment analysis and pathway annotation analysis using the DAVID database.The UHPLC-Q-Orbitrap HRMS technique was used to characterize the major chemical components of Zhenzhu Tongluo Wan.In addition,molecular docking,animal and cell experiments were performed to explore the mechanisms underlying the treatment of peripheral neuropathy with Zhenzhu Tongluo Wan.Results A total of 147 potential active ingredients and 333 intersection targets of Zhenzhu Tongluo Wan were screened.GO function and KEGG pathway analysis revealed that the major pathways involved include MAPK signaling pathway,NF-κB signaling pathway,PI3K-Akt signaling pathway,MAPK cascade and inflammatory response,among others.Based on high-resolution mass spectrometry analysis,33 major active ingredients were identified from Zhenzhu Tongluo Wan,focusing on those compounds with a Dgree value greater than the median.Molecular docking results further demonstrate good binding interactions between compounds such as luteolin,quercetin,tryptophan,arginine and kaempferol found inZhenzhu Tongluo Wan and key targets.Western blotting experiments showed that luteolin,quercetin and tryptophan could significantly inhibit the expression of MAPK8,P65 and TNF proteins in inflammatory cells.Animal experiments showed that Zhenzhu Tongluo Wan significantly improved hyperalgesia in rats with diabetic peripheral neuropathy,which may be related to the modulation of the expression of MAPK8,P65 and TNF proteins in the dorsal root ganglion.Conclusion Zhenzhu tongluo pills may play a role in the treatment of peripheral neuropathy by acting on the targets of MAPK8,P65 and TNF.This study lays a foundation for further research on the mechanism and pharmacodynamic substances of Zhenzhu Tongluo Wan in the treatment of peripheral neuropathy.

Objective This study aimed to systematically investigate the pharmacological substances and potential mechanisms of Zhenzhu Tongluo Wan in the treatment of peripheral nerve disorders.Methods Based on the TCMSP database and related literature,the active ingredients of Zhenzhu Tongluo Wan were determined.Target proteins for these active ingredients and those associated with peripheral neuropathy were predicted using the TCMSP,SwissTarget Prediction,GeneCard,OMIM,and DisGeNET databases.The intersection of these two sets of proteins was once considered a key target for the treatment of peripheral neuropathy.A protein-protein interaction(PPI)network of the intersection targets was constructed using the STRING database and analyzed using Cytoscape 3.9.1 software.Key targets obtained from the network were subjected to GO enrichment analysis and pathway annotation analysis using the DAVID database.The UHPLC-Q-Orbitrap HRMS technique was used to characterize the major chemical components of Zhenzhu Tongluo Wan.In addition,molecular docking,animal and cell experiments were performed to explore the mechanisms underlying the treatment of peripheral neuropathy with Zhenzhu Tongluo Wan.Results A total of 147 potential active ingredients and 333 intersection targets of Zhenzhu Tongluo Wan were screened.GO function and KEGG pathway analysis revealed that the major pathways involved include MAPK signaling pathway,NF-κB signaling pathway,PI3K-Akt signaling pathway,MAPK cascade and inflammatory response,among others.Based on high-resolution mass spectrometry analysis,33 major active ingredients were identified from Zhenzhu Tongluo Wan,focusing on those compounds with a Dgree value greater than the median.Molecular docking results further demonstrate good binding interactions between compounds such as luteolin,quercetin,tryptophan,arginine and kaempferol found inZhenzhu Tongluo Wan and key targets.Western blotting experiments showed that luteolin,quercetin and tryptophan could significantly inhibit the expression of MAPK8,P65 and TNF proteins in inflammatory cells.Animal experiments showed that Zhenzhu Tongluo Wan significantly improved hyperalgesia in rats with diabetic peripheral neuropathy,which may be related to the modulation of the expression of MAPK8,P65 and TNF proteins in the dorsal root ganglion.Conclusion Zhenzhu tongluo pills may play a role in the treatment of peripheral neuropathy by acting on the targets of MAPK8,P65 and TNF.This study lays a foundation for further research on the mechanism and pharmacodynamic substances of Zhenzhu Tongluo Wan in the treatment of peripheral neuropathy.

OBJECTIVE To establish a quantitative analysis method for the quality control components in Tenghuang jiangu capsules， and predict the possible action mechanism of the quality control components. METHODS Seven key quality control components in Tenghuang jiangu capsules were quantitatively analyzed by UHPLC-Q-Orbitrap HRMS. The “component-target” network was constructed based on network pharmacology， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and gene ontology （GO） function enrichment analysis were further conducted to find the key signaling pathways. RESULTS The average contents of succinic acid， hyperoside， gallic acid， kaempferol， naringin， naringenin and protocatechuic acid in 20 batches of Tenghuang jiangu capsules were 520.92， 67.67， 129.48， 4.74， 397.45， 5.66 and 376.62 μg/g， respectively. The results of network pharmacology showed that the 62 key target genes of the quality control components of the drug included AKT1， TNF， VEGFA， MMP9， PTGS2， etc. They were mainly enriched in cytokine receptor interaction， nuclear factor， tumor necrosis factor， interleukin 17， rheumatoid arthritis， Toll-like receptor and other signal pathways， involving inflammatory reaction， signal transduction， protein phosphorylation and other biological processes， kytoplasm， cell membrane and other cell components， as well as enzyme activity， energy activity and other molecular functions. CONCLUSIONS The established UHPLC- Q-Orbitrap HRMS method can be used for the quantitative analysis of the quality control components of Tenghuang jiangu capsule. Its quality control components may be mapped to inflammatory pathways related to bone diseases such as rheumatoid arthritis and Toll-like receptors through AKT1， TNF， VEGFA and other key targets， so as to play a therapeutic role.