AIM:To investigate the mechanism of flavonoid extract from Dracocephalum rupestre hance(DRHF)in the treatment of gouty arthritis through network pharmacology,molecular docking and animal experiment.METHODS:Literature re-trieval was used to explore the main active chemi-cal components and targets of DRHF.Gouty arthri-tis disease targets were obtained using Gene Cards and OMIM databases,and drug-disease intersect-ing targets were obtained using Wayne online tools.protein-protein interactions(PPI)and other related network diagrams were constructed using Cytoscape software.GO and KEGG enrichment analyses were performed on the shared intersect-ing targets using Metascape database.A rat model of gouty arthritis was established by Coderre meth-od;the swelling degree of ankle joint,gait behav-iour scores of rats were observed,and hematoxylin-eosin(HE)staining was performed.ELISA and real-time PCR were used to detect the key targets pre-dicted by the network pharmacology,and the ef-fects of DRHF on the molecular mechanism and key targets of gouty arthritis were observed.RESULTS:A total of 7 active compounds and 129 candidate targets for the treatment of GA were obtained,in-cluding IL-6,IL-1β,RELA,TNF,PPARG,etc.and the KEGG enrichment results suggested that DRHF may be involved in PI3K-Akt,TNF,IL-17 and other signal transduction pathways.Animal results:HE staining showed that the thickening of synovial tissue was not obvious in each administered group,and syno-vial cell proliferation and inflammatory cell infiltra-tion were significantly improved;compared with the normal group,the serum levels of TNF,IL-6,and IL-1β in the model group were significantly higher(P＜0.05),and the mRNA of PPARG,IL-6,and RELA in the synovial tissues were significantly high-er;compared with the model group,the levels of TNF,IL-6,and IL-1β were significantly lower(P＜0.05)in the low group of DRHF(0.45 g/kg)and high group of DRHF(0.9 g/kg),TNF,IL-6,IL-1β lev-els were significantly reduced(P＜0.05);PPARG,IL-6,RELA mRNA in synovial tissue were significantly reduced.CONCLUSION:DRHF inhibits IL-17/PI-3K/TNF signaling pathway by down-regulating the ex-pression of IL-6,PPARG and RELA mRNA,decreas-ing the levels of IL-6,IL-1β and TNF,and then treat-ing gouty arthritis.

AIM:To investigate the mechanism of flavonoid extract from Dracocephalum rupestre hance(DRHF)in the treatment of gouty arthritis through network pharmacology,molecular docking and animal experiment.METHODS:Literature re-trieval was used to explore the main active chemi-cal components and targets of DRHF.Gouty arthri-tis disease targets were obtained using Gene Cards and OMIM databases,and drug-disease intersect-ing targets were obtained using Wayne online tools.protein-protein interactions(PPI)and other related network diagrams were constructed using Cytoscape software.GO and KEGG enrichment analyses were performed on the shared intersect-ing targets using Metascape database.A rat model of gouty arthritis was established by Coderre meth-od;the swelling degree of ankle joint,gait behav-iour scores of rats were observed,and hematoxylin-eosin(HE)staining was performed.ELISA and real-time PCR were used to detect the key targets pre-dicted by the network pharmacology,and the ef-fects of DRHF on the molecular mechanism and key targets of gouty arthritis were observed.RESULTS:A total of 7 active compounds and 129 candidate targets for the treatment of GA were obtained,in-cluding IL-6,IL-1β,RELA,TNF,PPARG,etc.and the KEGG enrichment results suggested that DRHF may be involved in PI3K-Akt,TNF,IL-17 and other signal transduction pathways.Animal results:HE staining showed that the thickening of synovial tissue was not obvious in each administered group,and syno-vial cell proliferation and inflammatory cell infiltra-tion were significantly improved;compared with the normal group,the serum levels of TNF,IL-6,and IL-1β in the model group were significantly higher(P＜0.05),and the mRNA of PPARG,IL-6,and RELA in the synovial tissues were significantly high-er;compared with the model group,the levels of TNF,IL-6,and IL-1β were significantly lower(P＜0.05)in the low group of DRHF(0.45 g/kg)and high group of DRHF(0.9 g/kg),TNF,IL-6,IL-1β lev-els were significantly reduced(P＜0.05);PPARG,IL-6,RELA mRNA in synovial tissue were significantly reduced.CONCLUSION:DRHF inhibits IL-17/PI-3K/TNF signaling pathway by down-regulating the ex-pression of IL-6,PPARG and RELA mRNA,decreas-ing the levels of IL-6,IL-1β and TNF,and then treat-ing gouty arthritis.

This study aimed to investigate the effect and mechanisms of Ephedra Herb (EH) extract on adriamycin-induced nephrotic syndrome (NS), providing an experimental basis for the clinical treatment of NS. Hematoxylin and eosin staining, creatinine, urea nitrogen, and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function. The levels of inflammatory factors and oxidative stress were detected by kits. The levels of reactive oxygen species, immune cells, and apoptosis were measured by flow cytometry. A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS. The protein levels of apoptosis-related proteins and CAMKK2, p-CAMKK2, AMPK, p-AMPK, mTOR and p-mTOR in the kidneys were detected by Western blot. The effective material basis of EH extract was screened by MTT assay. The AMPK pathway inhibitor (compound C, CC) was added to investigate the effect of the potent material basis on adriamycin-induced cell injury. EH extract significantly improved renal injury and relieve inflammation, oxidative stress, and apoptosis in rats. Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury. Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR, which were blocked by CC. In sum, EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine may be one of the material bases of EH extract.
Rats ; Animals ; Doxorubicin/adverse effects* ; Nephrotic Syndrome ; AMP-Activated Protein Kinases/metabolism* ; Signal Transduction ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

Objective:To study the characteristics of the lens turbidity after long-term exposure to low intensity 635nm laser.Methods:Cluster sampling method was adopted to select 812 employees in a laser leveler workshop in a city of Guangdong Province from January 2014 to December 2018. They were divided into the control group, diffuse reflection (DR) group and direct vision (DV) group for retrospective observation and analysis of lens turbidity. The laser irradiation intensity of each group was investigated, the position and shape of lens opacity were analyzed, and the influencing factors were statistically analyzed with the repeated measurement data of dichotomy.Results:The laser irradiance and radiant exposure of DV group were between 0.72×10 -4 and 9.92×10 -4 mW/cm 2 and between 2.61×10 -2 and 1.53 J/cm 2, respectively. The subjects were mainly diagnosed with lens turbidity lesion, especially for the DV group. Most of lesions occurred in the pole and periphery of the anterior cortex. The lesions exhibited multipoint patterns with greyish white color. The turbidity rates in DV group (before work and work for 1, 2, 3 years) were 0%, 1.99% (8/402) , 4.98% (20/402) and 6.72% (27/402) , respectively, in the order of observation points. The statistical analysis of single factor effect showed that the turbidity rate was higher in DV group and higher in the second year in the DV group ( P<0.01) . Multi-factor analysis of the laser effect on the lens showed that the main effect between groups, between the observation point were statistically significant ( P<0.05) , but no statistical significance in the interaction between group×observation points ( P>0.05) . Conclusion:Lens turbidity lesion can be caused by long-term exposure to low intensity 635 nm laser, so the product safety classification should be strictly strengthened. It is necessary to strengthen the protection of laser photochemical damage in the production process.

Objective:To study the characteristics of the lens turbidity after long-term exposure to low intensity 635nm laser.Methods:Cluster sampling method was adopted to select 812 employees in a laser leveler workshop in a city of Guangdong Province from January 2014 to December 2018. They were divided into the control group, diffuse reflection (DR) group and direct vision (DV) group for retrospective observation and analysis of lens turbidity. The laser irradiation intensity of each group was investigated, the position and shape of lens opacity were analyzed, and the influencing factors were statistically analyzed with the repeated measurement data of dichotomy.Results:The laser irradiance and radiant exposure of DV group were between 0.72×10 -4 and 9.92×10 -4 mW/cm 2 and between 2.61×10 -2 and 1.53 J/cm 2, respectively. The subjects were mainly diagnosed with lens turbidity lesion, especially for the DV group. Most of lesions occurred in the pole and periphery of the anterior cortex. The lesions exhibited multipoint patterns with greyish white color. The turbidity rates in DV group (before work and work for 1, 2, 3 years) were 0%, 1.99% (8/402) , 4.98% (20/402) and 6.72% (27/402) , respectively, in the order of observation points. The statistical analysis of single factor effect showed that the turbidity rate was higher in DV group and higher in the second year in the DV group ( P<0.01) . Multi-factor analysis of the laser effect on the lens showed that the main effect between groups, between the observation point were statistically significant ( P<0.05) , but no statistical significance in the interaction between group×observation points ( P>0.05) . Conclusion:Lens turbidity lesion can be caused by long-term exposure to low intensity 635 nm laser, so the product safety classification should be strictly strengthened. It is necessary to strengthen the protection of laser photochemical damage in the production process.

Objective: To investigate the effects of astaxanthin on neuronal injury in hippocampus of rats after acute carbon monoxide poisoning(ACMP) and the relationship with NF-κB inflammatory signaling pathway. Methods: Male SD rats screened by water maze were randomly divided into three group(n=50): control group (NC group), CO poisoning group (COP group), CO poisoning+ astaxanthin group (AST group) . ACMP rat model was established by static inhaled exposure method. Meanwhile, rats in AST group were further given astaxanthin twice a day by gavage.At 1 day, 7 days, 14 days, 21 days and 28 days after CO poisoning (10 rats in each group were selected), the learning and memory abilities were evaluated by Morris water maze test. The pathological changes of the neurons in hippocampal CA1 region were observed by hematoxylin-eosin(HE) staining.The expression and activation of NF-κB in hippocampus were detected by immunoblotting and immunofluorescence, and the expression of TNF-α and IL-6 protein in hippocampus were examined by ELISA. Results: Morris water maze test showed that there were no significant difference in escape latency and crossing platform times between the three groups (P>0.05). Compared with NC group, the escape latency of COP group was prolonged at 14, 21 and 28 days after CO poisoning (t=-6.04, -6.28, -8.18, all P<0.05), and the number of crossing platform was decreased (t= 5.96, 7.85, 6.51, all P<0.05). Compared with the COP group, the escape latency of the rats in AST group at the 14, 21 and 28 days was shortened (t=4.74, 4.82, 5.98, all P<0.05), and the number of crossing platform was increased (t=-3.72, -4.45, -6.53, all P<0.05). Compared with NC group, the number of NF-κ B positive cells in CA1 area of hippocampus in COP group increased at every time point (t=-8.62, -18.00, -16.67, -11.15, -6.22, all P<0.05); the number of NF-κ B positive cells in CA1 area in AST group decreased at 7 d, 14 d, 21 d and 28 d after CO poisoning, the difference was statistically significant (t= 6.55, 6.96, 4.40, 4.17; all P<0.05). Western blot showed that the changes of NF-κB protein was similar to that of immunofluorescence. After 7 days of CO poisoning, the level of NF-κB protein in hippocampus of COP group was (1.44±0.08), it was higher than that of NC group (t=-20.07, P<0.05), while that of AST group was (0.68±0.10), it was lower than that of COP group (t=10.23, P<0.05). The results of Elisa showed that TNF-α and IL-6 in the hippocampus of COP group were higher than those of NC group at every time point(all P<0.05), while compared with COP group, TNF-α and IL-6 in AST group were lower (all P<0.05). After 7 days of CO poisoning, TNF -α in COP group ((39.04±5.29) pg/ml)was higher than that in NC group ((14.13±2.12) pg/ml) (t=-7.58, P<0.05); TNF -α in AST group ((25.77±3.31) pg/ml) was lower than that in COP group (t=3.69, P<0.05). After 7 days of CO poisoning, the level of IL-6 in COP group ((181.79±9.12) pg/ml)was higher than that in NC group ((73.12±11.04) pg/ml) (t=-8.24, P<0.05), and the level of IL-6 in AST group ((121.47±9.80) pg/ml) was lower than that in COP group (t=7.80, P<0.05). Conclusion The excessive inflammatory response which mediated by NF-κB signaling pathway is involved in ACMP-induced neuronal damage in hippocampus.Astaxanthin can down-regulate the expression of NF-κB inflammatory signaling pathway related proteins, and pave a way for the treatment of ACMP brain damage and cognitive dysfunction.

Objective To construct two DNA vaccines encoding Gag-Env fusion protein and Tat-Rev-Integrase(C-half)-Vif-Nef fusion protein derived from the first HIV-1 CRF01_AE isolate(AE2f) in Chi-na and to evaluate the immunogenicity in mice. Methods Two DNA vaccines were constructed by inserting the codon optimized and synthesized gag-env fusion gene and tat-rev-integrase(c-half)-vif-nef fusion gene de-rived from AE2f into mammalian expression vector pDRVISV1. 0, the generated DNA vaccines were desig-nated as pSVAE/GE and pSVAE/TRIVN, respectively, and their in vitro expression were determined by Western blot with transfected 293T cells. Mice were i. m. immunized with either pDRVI1.0 as mock control, pSVAE/GE or pSYAE/TRIVN for 4 times at two-week interval. Two weeks following the final im-munization, cellular responses to pool of HIV-1 Env, Gag, Tat, Rev, Intergrase, Vif and Nef peptides were evaluated by ELISPOT assay. Results The construction of DNA vaccine pSVAE/GE and pSVAE/TRIVN was validated by restriction enzyme digestion and bidirectional sequencing. Western blot showed a specific band at molecular mass 220×10~3 in lane of pSVAE/GE transfeeted 293T cell and a specific band at 95×10~3 in the lane of pSVAE/TRIVN. Both DNA vaccines mounted significant specific T cell responses with (3010 ± 566) SFC/10~6 splenocytes for DNA vaccine pSV AE/GE and (948 ± 737) SFC/10~6 spleno-cytes for DNA vaccine pSVAE/TRIVN, whereas the mock control of pDRVISV1.0 only raised marginal T cell responses. Conclusion Both pSVAE/GE and pSVAE/TRIVN were capable of expressing the inserted fusion immunogen genes and able to elicit vigorous cellular immune responses, therefore, these DNA vac-cines are highly immunogenic.