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ObjectiveTo investigate the effects of Yishen Daluo prescription （YSDL） on Ras homolog（Rho）/Rho-associated coiled-coil containing protein kinase（ROCK）signaling pathway in mice with experimental autoimmune encephalomyelitis （EAE） based on the silencing of β-arrestin1 gene. MethodSixty C57BL/6 female mice were randomly divided into a blank group， a model group， a virus group， a YSDL group， a virus + YSDL group， and a prednisone acetate group （hormone group）. The EAE model was induced in mice except for those in the normal group. Adeno-associated virus（AAV）solution （150 μL， 1×1011 vg·mL-1） was injected into the tail vein of each mouse in the virus group and the virus + YSDL group on the 4th day of immunization. Drugs were administered on the 8th day of modeling. Specifically， normal saline was given to the mice in the normal group，the model group，and the virus group at 10 mL∙kg-1， prednisone acetate suspension to those in the hormone group at 3.9 g∙kg-1，and YSDL to those in other groups at 20 g∙kg-1 for 14 consecutive days. The mice were weighed and scored every day. The neurological function scores of mice in each group were recorded every day after immunization. Hematoxylin-eosin （HE） staining was used to determine the inflammatory response and lesion location in the brain tissues and spinal cord tissues of mice. The protein expression of β-arrestin1，Ras homolog gene family member A（RhoA）， and Rho-associated coiled-coil forming protein kinase Ⅰ（ROCK Ⅰ） in spinal cord and brain tissues of EAE mice was determined by Western blot. ResultCompared with the model group， the virus group and the virus + YSDL group showed decreased neurological function scores （P<0.01），and the YSDL group also showed decreased neurological function scores（P<0.05）. HE results showed that there was obvious inflammatory reaction in the central nervous system （CNS） of the model group， which was alleviated to varying degrees in other groups compared with the model group. Western blot results showed that compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the spinal cord tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression levels of β-arrestin1， RhoA， and ROCKⅠ in the spinal cord tissues （P<0.01）. Compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the brain tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression level of β-arrestin1 in the brain tissues （P<0.01）， and the virus group and the YSDL group showed decreased protein expression levels of RhoA， and ROCKⅠ in the brain tissues （P<0.05）. Additionally， the virus + YSDL group and the hormone group showed decreased protein expression levels of RhoA and ROCKⅠ in the brain tissues （P<0.01）. ConclusionYSDL can improve the clinical symptoms of EAE mice and improve the inflammatory response of CNS. The mechanism is presumably attributed to the fact that YSDL inhibits the expression of β-arrestin1 in CNS，thereby reducing the expression of Rho/ROCK signaling pathway. Furthermore， YSDL may have a synergistic effect with the inhibition of β-arrestin1 gene expression.
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The academic levels and contents of papers published in China on random control tests of treatment of infantile mycoplasmal pneumonia with traditional Chinese medicine were assessed and analyzed according to the current treatment of infantile mycoplasmal pneumonia with traditional Chinese medicine and the problems existed in order to further improve its clinical treatment.
RESUMEN
Five field strains of infectious bronchitis virus (IBV) were isolated from suspected flocks from different time and different regions of Shanxi province,respectively,and characterized by a series of systematic identification assays,such as morphological observation by electron-microscope,interfering with the propagation of NDV,virus pathological role to chicken embryo,virus pathological role to SPF chickens,hemagglutination activity,physiscochemical,and reverse transcription polymerase chain reaction(RT-PCR).The results showed:The typical coronavirus which the spherical virions 60-120 nm in diameter and surface covered with spike like corona were observed under electron-microscope)The propagation of NDV strain was seriously interfered by the 5 isolates respectively;The embryonated chicken egg passages of the 5 isolates could dwarf with chicken embryos;The five isolates had no hemagglutination activity,but after treatment with 1% trypsin,it can agglutinate chicken red blood cell.The strains are sensitive to chloroform and ethyl ether.The SPF chickens which inoculated with the 5 isolates showed clinical sign and result in respiratory and kidney diseases,flower-steak,and swollen with severe urate deposition.The specific fragments of N gene of the 5 isolates were amplified by RT-PCR,respectiveiy.On the basis of all above mentioned results,the 5 isolates were classified as IBV.The study got a good preparedness for further study on molecular epidemioogy of the 5 IBV isolates.