Objective Using different concentrations of Coxsackievirus B3(CVB3)to infect young SD rats.To investigate the distribution of coxsackievirus B3(CVB3)in rat tissues and the immune response and inflammatory factors,to clarify the immunopathological mechanism of viral infection and provide an experimental basis for drug screening and efficacy evaluation.Methods Young SD rats(7 days old)were injected intraperitoneally with different doses of CVB3(TCID50=10-3.34/100 μL)and the proportions of lymphocyte subsets(CD4+,CD8+)in whole blood at days 4 and 8 were detected by flow cytometry.The CVB3 loads in the heart,liver,spleen,brain,kidney,and gastrointestinal tissues were detected by real-time fluorescence quantitative polymerase chain reaction,TNF-α and IFN-γ levels were detected by enzyme-linked immunosorbent assay,and histomorphologic changes were observed by hematoxylin and eosin staining.Results Different doses of CVB3 caused different degrees of diarrhea and decreased body mass in young rats.CVB3 was mainly distributed in the stomach,small intestine,large intestine,and stools,with the highest load in the large intestine and stools.The stock solution group(TCID50=10-3.34/100 μL)increased the proportion of CD8+T cells in the whole blood in young rats and decreased the CD4+/CD8+ratio(P＜0.05,P＜0.01).Compared with the nomal group high TNF-α and low IFN-γ expression were observed in the large intestine of young rats in the concentrate group(P＜0.05,P＜0.01),and submucosal edema and inflammatory cell infiltration were observed in the large intestine(cecum and rectum).There were no significant differences in the proportion of lymphocyte subsets,TNF-α and IFN-γ levels,and morphological changes in whole blood of young rats in the group 10-1,10-2,and 10-3(P＞0.05).Conclusions Different doses of CVB3 can induce infections in young SD rats.CVB3(TCID50=10-3.34/100 μL)causes pathological changes in the large intestine(cecum and rectum)in young rats,and high virus replication can increase levels of inflammatory factors and cause an imbalance of immune cells.CVB3 may have a unique pathogenic mechanism in young rats,providing a theoretical basis for developing evaluation strategies for drugs against CVB3 virus infections.

Objective To investigate the preventive effects of lidocaine administered through different routes on cardiovascular stress responses during anesthesia tracheal intubation. Methods Total 120 patients scheduled for elective surgery under general anesthesia were randomly divided into three groups: intravenous injection group （group IV）, throat spray group （group LJ）, and control group （group CT）, with 40 patients in each. Group IV received 50 mg of lidocaine via intravenous injection 1 minute before tracheal intubation. Group LJ received 50 mg of lidocaine sprayed into the pharyngeal cavity, glottis, and subglottic area. Group CT did not receive any treatment, and the remaining procedures were performed following the routine general anesthesia induction protocol. Heart rate （HR）, systolic blood pressure （SBP）, diastolic blood pressure （DBP）, and mean arterial pressure （MAP） were recorded at four time points: T₀ （before tracheal intubation）, T₁ （immediately after tracheal intubation）, T₂ （3 minutes after intubation）, and T₃ （5 minutes after intubation）. Statistical analysis of the data was performed using SPSS 22.0. Results There were no significant differences in HR at various time points within the group LJ. The changes in HR in the group IV and group CT were different statistically from those in the throat spray group. The blood pressure of patients in all three groups increased to varying degrees immediately after tracheal intubation, with the group CT showing particularly significant changes that differed significantly from both the group IV and the group LJ. The group LJ rapidly returned to levels close to those before intubation. Conclusion The preventive effects of lidocaine on stress responses during tracheal intubation were different depending on the route of administration. The inhibitory preventive effect of the throat spray method was superior to that of intravenous lidocaine, especially in preventing changes in heart rate.

Objective Using different concentrations of Coxsackievirus B3(CVB3)to infect young SD rats.To investigate the distribution of coxsackievirus B3(CVB3)in rat tissues and the immune response and inflammatory factors,to clarify the immunopathological mechanism of viral infection and provide an experimental basis for drug screening and efficacy evaluation.Methods Young SD rats(7 days old)were injected intraperitoneally with different doses of CVB3(TCID50=10-3.34/100 μL)and the proportions of lymphocyte subsets(CD4+,CD8+)in whole blood at days 4 and 8 were detected by flow cytometry.The CVB3 loads in the heart,liver,spleen,brain,kidney,and gastrointestinal tissues were detected by real-time fluorescence quantitative polymerase chain reaction,TNF-α and IFN-γ levels were detected by enzyme-linked immunosorbent assay,and histomorphologic changes were observed by hematoxylin and eosin staining.Results Different doses of CVB3 caused different degrees of diarrhea and decreased body mass in young rats.CVB3 was mainly distributed in the stomach,small intestine,large intestine,and stools,with the highest load in the large intestine and stools.The stock solution group(TCID50=10-3.34/100 μL)increased the proportion of CD8+T cells in the whole blood in young rats and decreased the CD4+/CD8+ratio(P＜0.05,P＜0.01).Compared with the nomal group high TNF-α and low IFN-γ expression were observed in the large intestine of young rats in the concentrate group(P＜0.05,P＜0.01),and submucosal edema and inflammatory cell infiltration were observed in the large intestine(cecum and rectum).There were no significant differences in the proportion of lymphocyte subsets,TNF-α and IFN-γ levels,and morphological changes in whole blood of young rats in the group 10-1,10-2,and 10-3(P＞0.05).Conclusions Different doses of CVB3 can induce infections in young SD rats.CVB3(TCID50=10-3.34/100 μL)causes pathological changes in the large intestine(cecum and rectum)in young rats,and high virus replication can increase levels of inflammatory factors and cause an imbalance of immune cells.CVB3 may have a unique pathogenic mechanism in young rats,providing a theoretical basis for developing evaluation strategies for drugs against CVB3 virus infections.

Objective To establish a model of indirectly induced respiratory tract infection with influenza A subtypes H1N1 and H3N2 in animals,to screen influenza virus hosts,and to provide theoretical support for the clinical control of influenza viruses.Methods Fifty BALB/c mice and 50 Hartley guinea pigs were randomly divided into five groups(10 animals/group for each species):normal control group,virus infects 1 group,virus infects 2 group,close transmission 1 group,and close transmission 2 group.Mice and guinea pigs in virus infects 1 and 2 groups were administered influenza A(H1N1)and influenza A(H3N2)viruses via nasal drip.For both virus infects 1 and 2 groups,animals were housed together with those in the close transmission group at a 1∶1 ratio on the following day.On day 7,the lung function,viral titer and viral load of the nasal tissue,trachea,and lung tissue of each group were measured,and pathological changes of the trachea and lung tissue of animals in the close transmission group were evaluated.Results In mice,the viral titers and viral loads of nasal,tracheal,and lung tissues of virus infects 1 and 2 and the closely transmitted groups 1 and 2 were significantly higher(P＜0.01),pathological scores of the trachea and lung tissues were significantly higher(P＜0.01),and the FVC and FEV20 of virus infects l and 2 groups were significantly lower(P＜0.01)than those in the normal control group.The nasal tissue,trachea and lung tissues of guinea pigs in virus infects 1 and 2 groups and close transmission groups 1 and 2 showed significantly higher viral titers and viral loads(P＜0.01),significantly higher trachea and lung histopathological scores(P＜0.01),and significantly lower FVC and FEV200(P＜0.01)than those of the normal control group.Conclusions In this study,influenza A subtypes H1N1 and H3N2 were used to indirectly induce respiratory tract infections in mice and guinea pigs for analyses of animal lung function,respiratory viral titers,viral load,and pathology.The animal models of the indirect transmission of influenza viruses in the respiratory tract had certain limitations;for example,influenza viruses were transmitted less efficiently among mice than among guinea pigs.The guinea pig model was stable.These findings confirm that guinea pigs are suitable hosts for efficient virus replication and transmission.

Objective To establish a model of indirectly induced respiratory tract infection with influenza A subtypes H1N1 and H3N2 in animals,to screen influenza virus hosts,and to provide theoretical support for the clinical control of influenza viruses.Methods Fifty BALB/c mice and 50 Hartley guinea pigs were randomly divided into five groups(10 animals/group for each species):normal control group,virus infects 1 group,virus infects 2 group,close transmission 1 group,and close transmission 2 group.Mice and guinea pigs in virus infects 1 and 2 groups were administered influenza A(H1N1)and influenza A(H3N2)viruses via nasal drip.For both virus infects 1 and 2 groups,animals were housed together with those in the close transmission group at a 1∶1 ratio on the following day.On day 7,the lung function,viral titer and viral load of the nasal tissue,trachea,and lung tissue of each group were measured,and pathological changes of the trachea and lung tissue of animals in the close transmission group were evaluated.Results In mice,the viral titers and viral loads of nasal,tracheal,and lung tissues of virus infects 1 and 2 and the closely transmitted groups 1 and 2 were significantly higher(P＜0.01),pathological scores of the trachea and lung tissues were significantly higher(P＜0.01),and the FVC and FEV20 of virus infects l and 2 groups were significantly lower(P＜0.01)than those in the normal control group.The nasal tissue,trachea and lung tissues of guinea pigs in virus infects 1 and 2 groups and close transmission groups 1 and 2 showed significantly higher viral titers and viral loads(P＜0.01),significantly higher trachea and lung histopathological scores(P＜0.01),and significantly lower FVC and FEV200(P＜0.01)than those of the normal control group.Conclusions In this study,influenza A subtypes H1N1 and H3N2 were used to indirectly induce respiratory tract infections in mice and guinea pigs for analyses of animal lung function,respiratory viral titers,viral load,and pathology.The animal models of the indirect transmission of influenza viruses in the respiratory tract had certain limitations;for example,influenza viruses were transmitted less efficiently among mice than among guinea pigs.The guinea pig model was stable.These findings confirm that guinea pigs are suitable hosts for efficient virus replication and transmission.

Objective A model for studying oral ulcers induced by betel nut-extract was constructed in rats.Changes in the structure and diversity of oral flora were observed to explore the involvement of oral flora and local inflammatory factors in the pathogenesis of oral ulcers induced by betel nut-extract and to provide theoretical support for the prevention and treatment of oral ulcers in the clinic.Methods Thirty SD rats were randomly divided into normal,model and intervention groups(Guilin watermelon cream,8 mg/d for 7 days),with 10 rats/group.The oral mucosa of rats was subcutaneously injected with 10 g/mL of betel nut-extract to generate an oral ulcer model.The histomorphological changes were observed,and ulcer area and ulcer scores were assessed.Local oral tissue tumor necrosis factor-α(TNF-α),interleukin(IL)-2 and IL-8 levels were determined.Oral mucosal tissues were sampled for HE staining and analyzed for the structural distribution of oral flora and the diversity of microbial communities using high-throughput sequencing method.Results Compared with rats in the normal group,those in the model group had an increased ulcer area,significantly increased ulcer scores(P＜0.01),and significantly increased levels of TNF-α,IL-2 and IL-8 in the oral mucosal tissues(P＜0.01).The amount Streptococcus(P＜0.05)and Veillonella(P＜0.001)in the oral saliva of the model group rats was significantly reduced.The model group rats showed oral mucosal epithelial cell hyperplasia or focal necrosis,mucosal lamina propria edema,and hemorrhage accompanied by mass neutrophil and monocyte infiltration.Compared with the model group rats,the intervention group rats had significantly reduced ulcerated area(P＜0.05,P＜0.01)and ulcer scores(P＜0.05).And oral mucosal tissue levels of TNF-α(P＜0.01),IL-2(P＜0.05)and IL-8(P＜0.05),as well as significantly increased Streptococcus(P＜0.001)and Veillonella(P＜0.01)and significantly reduced Staphylococcus(P＜0.01)in the oral saliva.The degree of lesions in the oral mucosal tissues was significantly improved in the intervention group.Conclusions Betel nut-extract can be used to successfully reproduce a rat model of oral ulcer,and it is speculated that the development of oral ulcers after exposure to betel nut-extract may be related to an imbalance in the oral flora and local tissue inflammatory mediators.

Ferrets offer an advantage in nonclinical studies of anti-infective drugs because of their ability to be infected with and spread pathogenic microorganisms,especially viral strains,without the need for host adaptation.Additionally,the clinical symptoms exhibited by infected ferrets are very similar to those of humans.Although ferrets play a very important role in the research and development of antiviral drugs,the scope of their application remains limited.This may be related to the lack of corresponding national standards for laboratory animal feeding and application of ferrets as well as the lack of specific diagnostic and detection reagents.This paper summarizes the characteristics of ferrets as infectious disease models with a summary and analysis of the application direction of ferrets in anti-infective drug research.Our aim is to promote further standardization of the use of ferrets.

Objective This study established a model of invasive Aspergillus niger lung disease in immunosuppressed rats to provide theoretical support for the pharmacodynamic evaluation of anti-invasive pulmonary aspergillosis drugs and mechanism studies.Methods Sixty SD rats were randomly divided into a normal control group;cyclophosphamide control group,and cyclophosphamide+fungal infection low,medium,and high dose groups,with 12 animals in each group.General clinical observations were performed daily,and the serum levels of immunoglobulin(Ig)G and IgM and galactomannan(GM)were detected by ELISA on the 3rd and 7th days of modeling.Simultaneously,the ratio of CD4+and CD8+cells,content of white blood cells(WBCs)and neutrophils(Neu)in peripheral blood,the Aspergillus niger load in alveolar lavage,and morphological changes to rat lung tissue were observed.Results Rats in the cyclophosphamide control and cyclophosphamide+fungal infection groups showed reduced voluntary activity and erect hair after modeling,and rats in the cyclophosphamide+fungal infection group also had shortness of breath and audible wet rhonchi in the lungs.Compared with the normal control group,rats in the cyclophosphamide control group showed significant reductions in the levels of CD4+,WBC,Neu,IgG,and IgM in the blood,and their proportion of CD8+cells was significantly higher(P＜0.05,P＜0.01).Compared with the cyclophosphamide control group,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly reduced blood levels of IgG,IgM,and CD4+cells(P＜0.05,P＜0.01);while the cyclophosphamide+fungal infection low-,medium-,and high-dose groups had significantly reduced blood levels of WBC and Neu(P＜0.05,P＜0.01).Additionally,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly increased blood CD8+cells(P＜0.05,P＜0.01),Blood GM levels and the alveolar lavage Aspergillus niger load were significantly increased in rats in the cyclophosphamide+fungal infection low-,medium-,and high-dose groups compared with the cyclophosphamide control group(P＜0.05,P＜0.01).The lung tissues of the cyclophosphamide+fungal infection low-,medium-,and high-dose groups showed mycelial distribution and destruction of alveolar epithelium,increase of bronchial epithelial cup cells in the alveoli,and infiltration of inflammatory cells,and the degree of lesions was positively correlated with the modeling dose.Conclusions In this study,we used Aspergillus niger combined with cyclophosphamide immunosuppressant to construct a model of invasive Aspergillus niger lung disease.The duration of the disease was positively correlated with the concentration of bacterial fluid and modeling time,confirming that cellular immunity plays an important role in the pathogenesis of the disease.At the same time,Ig can also affect the development of invasive pulmonary aspergillosis,and it is speculated that the pathogenesis may be related to the level of Ig produced by humoral immunity.

OBJECTIVE To explore the role of Yes-associated protein(YAP)in epidermal stem cell(EPSC)differentiation after ionizing radiation(IR).METHODS ① A punch was used to induce skin injuries on the back of mice.The IR group received localized irradiation with 60 Co γ-rays,while the normal control group did not.Samples were collected at 0,1,3,6,9,and 12 d for RNA and protein extraction.Western blotting was used to detect changes in YAP protein expressions during wound healing.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to assess the mRNA levels of Yap and its downstream target genes,connective tissue growth factor(Ctgf),and cysteine-rich protein 61(Cyr61).② EPSCs were exposed to 60 Co γ at a dose of 4 or 8 Gy,while the control group was not irradiated.Cells were collected to detect the levels of YAP protein via Western blotting.Cells were collected at 4,12,24,and 36 h post-IR to assess the levels of YAP mRNA by RT-qPCR.③ Short hairpin RNA(shRNA)was used to establish stable YAP knockdown cell lines,and the knockdown efficiency of sh YAP was verified by Western blotting.RT-qPCR was then performed to detect the impact of YAP knockdown on mRNA levels of K1 and K10 after IR.RESULTS① Compared with the control group,the YAP protein level in the IR group during wound healing was significantly reduced(P<0.05,P<0.01),so were the mRNA levels of Yap and its downstream target genes Ctgf and Cyr61(P<0.05,P<0.01).② Compared to the cell control group,the mRNA and protein levels of YAP in the IR group cells were significantly reduced(P<0.01).③ In the sh YAP cells,the YAP protein level was significantly reduced(P<0.01).Furthermore,the mRNA levels of K1 and K10 were significantly decreased after IR in sh YAP cells(P<0.01).CONCLUSION YAP can regulate EPSC differentiation in wound healing after IR.

Epilepsy is one of the common diseases of the nervous system,which is characterized by repeated epilep-tic seizures caused by abnormal discharge of brain neurons.Brivaracetam is an analogue of the third-generation antiepilep-tic drug levetiracetam and exerts a therapeutic effect by binding to synaptic vesicular protein 2A.This article reviews the disease of epilepsy and the mechanism of action,pharmacokinetic characteristics,clinical efficacy,safety,and adverse reactions of brivaracetam in epilepsy,in order to provide a better choice of drugs for the treatment of epilepsy.