Objective To explore the effect of salbutamol combined with Rho associated coiled-coil forming protein kinase (ROCK) inhibitor Y-27632 on airway smooth muscle and to find a new way for drug treatment of asthma. Methods Pig tracheal smooth muscle tissue strips were prepared, and after treatment they were divided into an electrical stimulation group (Fmax, 50%Fmax) and a blank group. The smooth muscle tissue strips were quickly frozen to determine the expression level of Rock-Ⅱ and the phosphorylation level of MLC20. The Fmax and 50%Fmax electrical stimulation groups were divided into a blank group, a salbutamol group, a Y-27632 group, and a salbutamol combined with Y-27632 group according to different intervention drugs. The relaxation of smooth muscle strips was observed. Results In the blank group, 50%Fmax group and Fmax group, the expression level of Rock-Ⅱ and the phosphorylation of MLC20 in smooth muscle tissue showed an increasing trend, with statistically significant differences (P<0.05). In the 4 subgroups of the 50%Fmax group intervention with different drugs (blank group, salbutamol group, Y-27632 group, salbutamol plus Y-27632 group), the diastolic ratio smooth muscle tissue strips showed an increasing trend. When the time reaches 10 min, the diastolic ratios were 0.7%, 2.5%, 6.0%, and 15.0%. the diastolic ratios were 1.8%, 4.5%, 7.5%, and 21.0% at the time of 20 min. the diastolic ratios were 1.9%, 7.5%, 7.9% and 22.0% at the time of 40 min. the diastolic ratios were 2.0%, 8.0%, 8.8%, and 22.5% at the time of 60 min. In the four subgroups of the Fmax electrical stimulation group, the relaxation ratio of smooth muscle tissue strips also showed an increasing trend. When the time reaches 10 min, the diastolic ratios were 1.0%, 3.0%, 7.0%, and 17.0%. the diastolic ratios were 2.6%, 5.5%, 9.0%, and 24.0% at the time of 20 min. the diastolic ratios were 2.8%, 9.0%, 9.5%, and 27.5% at the time of 40 min. diastolic ratios were 2.9%, 10.5%, 10.5%, and 28.0% at the time of 60 min. The analysis of difference between groups showed that at the same time, the diastolic ratio of smooth muscle in salbutamol combined with Y-27632 group was significantly higher than that in salbutamol alone group and Y-27632 group (P<0.05). In addition, the smooth muscle diastolic ratio of combined intervention was also better than the the mathematical sum effect of both single drug intervention (P<0.05). Conclusions The contractility and intensity of smooth muscle are positively correlated with the expression level of ROCK and the phosphorylation level of MLC20. Salbutamol combined with Y-27632 can enhance the relaxation of porcine airway smooth muscle, which may have a synergistic effect.
Objective To investigate the effect of the anterior cruciate l igament (ACL) rupture on the biomechanics of the lateral collateral l igament (LCL). Methods The specimens in this experiment were knee joints from 6 normal fresh adult male cadavers which was donated voluntarily, aged of 26-35 years with an average of 31.4 years. The knee joints were dissymmetry with 3 in left and right sides respectively. At first, all the 6 specimens lying on the self-made movement tooting, whose LCL straining were measured by strain gauges at different angles (0, 30, 60 and 90°) under axial loads of 400 N by e-testing machine for simulation of the normal knee joint force, were regarded as the intact ACL group. Then, the ACL in all 6 specimens were cut off completely as the deficiency group and did the same step. Results The strain of the LCL were as follows at 0, 30, 60 and 90°: (0.00 ± 1.63), (—17.2 ± 8.57), (—24.00 ± 4.80) and (26.50 ± 4.65) με in intact ACL group; (0.75 ± 8.22), (— 52.75 ± 3.33), (24.30 ± 4.99) and (26.30 ± 4.27) με in deficiency group. There was no significant difference at 0° and 90° flexion (P gt; 0.05), but significant difference at 30° and 60° flexion (P lt; 0.05) between the two groups. Conclusion The rupture of the ACL has significantly effect on the strain of the LCL which suffering abnormal load in the state at 30° and 60° flexion. At 30° flexion the relaxation of the LCL increased, which means the possibil ity of the injury of the LCL is rare; and at 60° flexion the LCL become very tense and thereby at the high risk of instabil ity.
To enhance the quality and transparency of oncology real-world evidence studies, the European Society for Medical Oncology (ESMO) has developed the first specific reporting guidelines for oncology RWE studies in peer-reviewed journals "the ESMO Guidance for Reporting Oncology Real-World Evidence (GROW)". To facilitate readers understanding and application of these reporting standards, this article introduces and interprets the development process and main contents of the ESMO-GROW checklist.
Objective To explore the potential molecular mechanism of Rhodiola crenulata (RC) for type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) by network pharmacology and molecular docking. Methods The target genes of T2DM and AD, the effective active components and targets of RC were identified through multiple public databases during March to August, 2022. The main active components and core genes of RC anti T2DM-AD were screened. The key genes were enrichment analyzed by gene ontology function and Kyoto gene and Kyoto Encyclopedia of Genes and Genomes. AutoDock Vina was used for molecular docking and binding energy calculation. Results A total of 5189 T2DM related genes and 1911 AD related genes were obtained, and the intersection result showed that there were 1418 T2DM-AD related genes. There were 48 active components of RC and 617 corresponding target genes. There were 220 crossing genes between RC and T2DM-AD. The main active components of RC anti T2DM-AD included kaempferol, velutin, and crenulatin. The key genes for regulation include ESR1, EGFR, and AKT1, which were mainly enriched in the hypoxia-inducible factor-1 signal pathway, estrogen signal pathway, and vascular endothelial growth factor signal pathway. The docking binding energies of the main active components of RC and key gene molecules were all less than −1.2 kcal/mol (1 kcal=4.2 kJ). Conclusions RC may play a role in influencing T2DM and AD by regulating the hypoxia-inducible factor-1 signaling pathway, estrogen signaling pathway, and vascular endothelial growth factor signaling pathway.
ObjectiveTo explore the metabolic changes during the differentiation of 3T3-L1 adipocytes caused by the treatment of the transient receptor potential vanilloid 4 (TRPV4)-specific agonist GSK1016790A basing on ultra-performance liquid chromatography-mass spectrometry technology. MethodsMouse 3T3-L1 cells were treated with GSK1016790A at different concentrations (0.1, 1, and 10 μmol/L), and the effect of drugs on cell proliferation was detected by cell counting kit-8 method. A mature adipocyte model was constructed, and GSK1016790A was used to activate TRPV4 channel protein activity and verify the expression levels of TRPV4 and triglycerides. Cell metabolites were collected for metabolomic studies, differential metabolites were screened between groups, and related metabolic pathways were analyzed. Results After GSK1016790A intervened in mature adipocytes, the expression levels of TRPV4 mRNA and triglycerides in cells were significantly upregulated (P<0.05). Metabolomics detection found that GSK1016790A screened a total of 45 differential metabolites such as 2-amino-1,3,4-octadecanetriol, linoleic acid, sphingosine, sphinganine, sn-glycerol-3-phosphate and uridine, mainly involving 13 possible metabolic pathways such as sphingolipid metabolism and biosynthesis of unsaturated fatty acids. Conclusion GSK1016790A may promote adipogenesis in adipocytes by activating TRPV4 channel protein activity, and at the same time participate in regulating metabolic pathways such as the biosynthesis of unsaturated fatty acids pathway and sphingolipid metabolism pathway, affecting lipid metabolism in adipocytes.