The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

Protein aggregation plays an essential role in the development of cataracts.In recent years,gene sequencing and other molecular biological techniques have been widely used in the study of protein aggregation,whose intrinsic mecha-nisms have been gradually elucidated.Gene mutations,isomerization and covalent modifications,compound and ionic in-teractions,and protein interactions are all causes of protein aggregation.This article focuses on the above four causes to review the research progress on the effect of protein aggregation in the development of cataracts,so as to provide informa-tion and reference for subsequent studies.

In order to understand the difference and expression of genes in CEK cells before and af-ter baicalin interferes with IBV,further reveal and analyze the mechanism of baicalin interfering IBV replication in CEK cells in vitro.After IBV infected CEK cells for 2 h,9.75 mg/L baicalin liq-uid was added to interfere with CEK cells,which was recorded as the baicalin(H-IBV)group,and three replicate wells were set in the control group and the IBV(IBV)infection group.After 36 h culture,cell samples were collected and subject to transcriptome for sequencing.The results showed that there were 102 differentially expressed genes in H-IBV group compared with IBV in-fection group,among which 48 genes weresignificantly up-regulated and 54 genes were significantly down-regulated.Through functional annotation in GO and KEGG databases,it was found that dif-ferentially expressed genes were mainly annotated in biological processes such as cellular proces-ses,biological regulation,metabolism,and secondary pathways such as viral infectious diseases,signal transduction and interaction.Retinol metabolism pathway,phospholipid transfer to mem-brane,IL-27 mediated signaling pathway,MDA5/RIG-I and Toll-like receptor signaling pathway were significantly enriched in CEK cells,and the production of type Ⅰ interferon and interferon al-pha and the process of antiviral infection were also positively regulated.There were more differen-tial genes enriched in nucleic acid catalysis,immune system,and reaction,and interbiological reac-tion.Through the STRING network interaction map,it was found that most immune-related genes could form a 36-node interaction network centered on IRF7,TLR3 and STAT1.Therefore,com-pared with IBV group,the differentially expressed genes after baicalin treatment were mainly an-notated and enriched in the biological process,and the immune system and response were en-hanced,mainly through the positive regulation of IRF7 in the MDA5/RIG-I receptor signaling pathway and the inhibition of TLR3 signal transduction in the Toll-like receptor signaling path-way.Positive regulation of IL-27 mediated pathway and regulation of JAK-STAT signaling path-way were supplemented by activation of the expression of IRF7,TLR3,STAT1 and other related genes,and interaction with corresponding downstream proteins to promote the expression of IFN-α and regulatory cytokines,coupled with negative regulation of viral(defense)response and viral processes.Thus,baicalin interferes with IBV replication in CEK cells.

ObjectiveTo observe the effects of electroacupuncture on IL-1β in brain tissue of rats after total cerebral ischemia-reperfusion.MethodsThirty male Wistar rats were randomly divided into the sham operation group, model group and acupuncture group, in each group. In acupuncture group and model group, 4-vessel-occlusion method was used to make a rat model of cerebral ischemia reperfusion. Acupuncture group used electroacupuncture on points Quchi and Zusanli, 6 hours after reperfusion. The rats of each group were killed and ELISA was used for the measurement of IL-1β in brain tissues.ResultsIL-1β in brain tissue in model group increased significantly compared with sham operation group and acupuncture group(P<0-001). No significant differences were found between sham operation group and acupuncture group (P>0-05).ConclusionAcupuncture can lower IL-1β content of brain tissue of rats with brain ischemia/reperfusion injury.

In order to remove invasive plasmids from Shigella flexneri 2a 2457T and Shigella sonnei S7 based on the principle of plasmid incompatibility Ori and inc genes were amplified by PCR from S. flexneri 2a invasive plasmids, and then they were cloned into plasmid pMD18 T, resulting recombinant plasmids pMDori and pMDinc After S flexneri 2a 2457T and S sonnei S7 were transformed with pMDori or pMDinc respectively Invasive plasmids were removed from S flexneri 2a 2457T and S sonnei S7