To rapidly detect porcine rotavirus(PoRV),a one-step detection method was developed based on recombinase-aided amplification(RAA)combined with lateral flow dipstick(LFD)tech-nology.Specific primers and probes were designed according to the conserved sequence of the PoRV VP6 gene,and the amplification system was optimized.The specificity,sensitivity,and re-peatability of the method were thoroughly tested,and its clinical application was evaluated.The re-sults demonstrated that the method could amplify PoRV nucleic acid at 37 ℃ in just 15 min with a minimal detection limit of 22.3 copies/μL.No cross-reactivity was observed with other common diarrhea viruses.The total coincidence rate between RT-RAA-LFD and RT-PCR was 97.0％,with a Kappa coefficient of 0.93(K＞0.75).In summary,the RT-RAA-LFD detection method estab-lished in this study exhibits excellent specificity,sensitivity,and stability,characterized by rapid detection speed and visualization,making it suitable for on-site rapid detection.This method pro-vides a novel technical approach for the rapid diagnosis and epidemiological investigation of PoRV.

To rapidly detect porcine rotavirus(PoRV),a one-step detection method was developed based on recombinase-aided amplification(RAA)combined with lateral flow dipstick(LFD)tech-nology.Specific primers and probes were designed according to the conserved sequence of the PoRV VP6 gene,and the amplification system was optimized.The specificity,sensitivity,and re-peatability of the method were thoroughly tested,and its clinical application was evaluated.The re-sults demonstrated that the method could amplify PoRV nucleic acid at 37 ℃ in just 15 min with a minimal detection limit of 22.3 copies/μL.No cross-reactivity was observed with other common diarrhea viruses.The total coincidence rate between RT-RAA-LFD and RT-PCR was 97.0％,with a Kappa coefficient of 0.93(K＞0.75).In summary,the RT-RAA-LFD detection method estab-lished in this study exhibits excellent specificity,sensitivity,and stability,characterized by rapid detection speed and visualization,making it suitable for on-site rapid detection.This method pro-vides a novel technical approach for the rapid diagnosis and epidemiological investigation of PoRV.

Aim To explore the mechanism of E.coli heat-labile enterotoxin B subunit(LTB)as adjuvant by analysis of cellular proteins interacting with LTB. Methods Whole cell proteins were purified from RAW 264.7 cell after treated with LTB or NaCl 12 h, respectively.The cellular proteins were interacted with LTB and the interacting proteins were purified by pull-down assay and identified by mass spectrography.The LTB interaction proteins were conformed with Western blot and immunofluorescence assay.Results 25 LTB interaction proteins were found,and their interaction network was mapped;four proteins (Jup,Dsp,Ddx5 and Vimentin)were indicated to be related with LTB adjuvant activity;immunofluorescence assay indicated that GM130 interacted with LTB,however,Vimentin had no interaction with LTB in vivo.After treated by LTB,the expression of β-actin was upregulated obvi-ously in RAW 264.7 cell,whereras,Hspd1 did not show any change.Conclusions LTB exerts adjuvant activity through binding to GM1 of immune cells,cau-sing endocytosis and transporting to the Golgi apparatus by vesicles.Then LTB might bind to Jup and affect TCF/LEF activity,regulating the expression of Bcl 2, IL-6,and Runx3.The result is promoted T cell and B cell proliferation,differentiation and activation by se-cretion of cytokines and immunoglobulins.

Aim Tostudytheeffectsofferulicacid (FA) on doxorubicin (DOX) induced cellular injury inH9c2ratmyocardialcells.Methods H9c2cells were treated with 1μmol·L-1 DOX treated for 24 h to establish a myocardial injury model. 10, 20, 40μmol ·L-1 FA was added 2 h before DOX treatment. Cell viability was measured by cell counter kit ( CCK-8 ) . Morphological changes were observed by phase contrast microscope. LDH, CK, MDA, SOD levels were detec-ted by biochemical kits. Intracellular level of reactive oxygen species ( ROS) was examined by DCF-DA stai-ning with flow cytometry. Cellular apoptosis was detec-ted by AO-EB staining and DNA agarose gel electro-phoresis. The expression of caspase-3, Bax, Bcl-2 was evaluatedbyWesternblot.Results Exposureof H9c2 cells to DOX led to decrease in cell viability, in-crease in stress and apoptosis. FA pre-treatment im-proved cell viability in a dose-dependent manner, at-tenuated leakage of LDH and CK, and reversed mor-phological changes induced by DOX. FA suppressed DOX-induced oxidative stress as evidenced by reducing ROS and MDA generation and increasing SOD enzyme activity. FA depressed myocardial apoptosis by down-regulating pro-apoptotic protein caspase-3 and Bax, whereas up-regulating apoptosis inhibitory protein Bcl-2.Conclusions FAhasaprotectiveeffectonDOX-induced injury in H9c2 cells. This protection may re-sult from inhibition of myocardial oxidative stress and apoptosis.

AIM: To investigate antitumor effect of allotri-tridecyl diethylamine (D-108) in vivo and in vitro. METHODS: The cytotoxic effects of D-108 on various tumor cell lines, human gingival fibroblast and marrow stromal cell cultured in vitro were determined with trypan blue dye exclusion test and MTT method. The acute toxicity of mice by administration of D-108 was evaluated by Bliss method. At a tolerable dose level, D-108 was administrated to treat transplanted solid tumor U14, and tumor weight inhibition was observed. Apoptosis morphological transformation of HL 60 cell induced by D-108 was detected by the Giemsa staining. RESULTS: The cytotoxic effects in vitro of D-108 on various tumor cell lines (IC_ 50 : 0.22 to 2.19 mg?L~ -1 ) were more powerful than both human gingival fibroblast and marrow stromal cell (IC_ 50 : 5.55 and 3.57 mg?L~ -1 ). LD_ 50 of D-108 was 36.49 mg?kg~ -1 (mice, i.g.). D-108 inhibited in vivo growth of implanted solid tumor U14 of mice effectually. The inhibition rate of tumor weight of D-108 (100 mg?kg~ -1 ?d~ -1 i.g.) was 45.27 %. HL 60 cell appearanced typical apoptosis morphological transformation induced by D-108. CONCLUSION: D-108 had obvious antitumor activity in vivo and in vitro and little toxicity. D-108 could induce the apoptosis of HL 60 cell.

AIM:To study the effect of Exendin-4 on the glucose tolerance and serum glucose level in normal animals. METHODS: The fasting blood glucose concentration was tested at 0,1,2,3,4 h and 2,4 week after the first administration of Exendin-4 (0.1, 0.3, 0.9 g/kg, 4 weeks, qd) in Wistar rats ,taking insulin as positive control. Before intragastric administration 2.5 g/kg glucose, Exendin-4 (0.2, 0.6, 1.8 g/kg) were subcutaneously injected, then the fasting blood glucose concentration was tested at 0.5, 1, 2 h after the glucose loading. After hypodermic administration of Exendin-4 (0.2, 0.6, 1.8 g/kg), half of the mice were subcutaneously administrated 2.5 g/kg glucose 15 min later, and the insulin was tested at the end of the experiment. RESULTS:Exendin-4 could not significantly change the fasting blood glucose concentration at different times. The fasting blood glucose concentration was significantly decreased after glucose loading by administration Exendin-4. Exendin-4 could increase the serum insulin concentration remarkably after glucose loading and could not change much without glucose loading. CONCLUSION: The results suggest that the blood glucose regulation of Exendin-4 was related to the concentration of glucose.