Objective:To investigate the effects of influenza A virus(IAV)on macrophages based on expression of cytokines mediated by Janus kinase/signal transducer and activator of transcription(JAK/STAT)pathway,and to further explore the intervention effects of Ma Xing Shigan decoction drug containing serum.Methods:RAW264.7 was divided into control group,JAK/STAT signal pathway activator group,inhibitor group,model group,oseltamivir group,antiviral granule group and Ma Xing Shigan decoction group.Cell samples were collected after the intervention of IAV and subsequent treating of drug-containing serum for 24 and 48 hours.Secretion levels of IL-1β and CXCL2 in supernatant were detected by ELISA.mRNA expression levels of influenza virus Nuclear Pro-tein(NP),JAK1/2 and STAT1 of cells were detected by RT-qPCR.Protein expression levels of JAK1/2 and STAT1 were detected by Western blot,and protein expression levels of p-STAT1 was detected by immunofluorescence.Correlation between protein expression levels of p-STAT1 and secretion of IL-1β and CXCL2 were evaluated by Pearson correlation analysis.Results:Secretion levels of IL-1β and CXCL2,mRNA expression levels of NP,JAK1/2,STAT1,protein levels of JAK1/2,STAT1 and p-STAT1 were increased,and protein levels of p-STAT1 was positively correlated with the secretion of IL-1β and CXCL2.Ma Xing Shigan decoction could down-regulate secretions of IL-1β and CXCL2,mRNA expression levels of NP,JAK1/2 and STAT1,protein expression levels of JAK1,JAK2,STAT1 and p-STAT1.Conclusion:Activation of JAK1/2-STAT1 signal pathway and imbalance of inflammatory factors may be one of the molecular mechanisms of immunopathological damage of macrophages induced by IAV.Ma Xing Shigan decoction may alle-viate pathogenic effects of IAV by regulating this signal pathway.

Objective:To explore the effect of Simo decoction improving the low duodenal inflammation and protecting the du-odenal mucosal barrier in rats with functional dyspepsia(FD).Methods:Sixty SD rats were randomly divided into control group(10 rats)and model group(50 rats),and the modeling rats were prepared by multivariate intervention method.After successful modeling,the modeling rats were randomly divided into model group,Simo decoction high-dose,medium-dose,low-dose group and mosapride group,with 10 rats in each group.The high-dose,medium-dose,and low-dose groups of Simo decoction were ga-vage given 5.62 g/kg,2.81 g/kg,and 1.40 g/kg,respectively,and the mosapride group was gavage given 0.305 mg/kg of mosapride,and the control group and model group were gavage given the same amount of distilled water for 14 days.The body weight of rats was observed;gastric emptying rate and small bowel propulsion rate were measured;transmission electron microscopy was used to observe the morphology of duodenal epithelial cells;ELISA detected serum levels of IL-17A and IL-22;Western blot and immunohistochemis-try were used to detect the expressions of protease-activated receptor 2(PAR-2)and tight junction protein(ZO-1,claudin-1)in the duodenum.Results:Compared with the control group,the body weight,gastric emptying rate and small intestinal propulsion rate of the model group were significantly reduced(P<0.01),transmission electron microscopy showed widening of the duodenal epithelial cell space,serum IL-17A and IL-22 levels were significantly increased(P<0.01),the expression of PAR-2 in duodenal tissue was in-creased,and the expressions of ZO-1 and claudin-1 were downregulated(P<0.01).Compared with model group,the gastric emptying rate and small intestinal propulsion rate in Simo decoction high-dose,medium-dose and mosapride group were increased(P<0.05,P<0.01),the contents of IL-17A and IL-22 in serum decreased(P<0.05,P<0.01),and the expression of PAR-2 in duodenal tissues was down-regulated,the expressions of ZO-1 and claudin-1 was significantly increased(P<0.01).The low-dose group of Simo soup could improve weight loss(P<0.01),reduce IL-17A content and PAR-2 expression,and increase ZO-1 and claudin-1 expression(P<0.05,P<0.01),while the effect on other indexes was not obvious.Conclusion:Simo decoction may reduce low-grade duodenal in-flammation to repair the mucosal barrier by down-regulating the levels of IL-17A and IL-22 and the expression of PAR-2,and up-regu-lating the expression of ZO-1 and claudin-1,so as to exert the effect of FD treatment.

Objective:To investigate the effect of bronchoalveolar lavage fluid(BALF)cytology examination combined with lymphocyte immunoassay on clinical outcome of critically ill patients with stroke-associated pneumonia(SAP).Methods:One hundred and fifty-six critically ill patients with SAP admitted to Department of Respiratory and Critical Care Medicine of the Second People's Hospital of Weifang from September 2021 to June 2022 were selected,and randomly divided into a study group and a control group,with 78 cases in each group.Control group was treated with conventional pulmonary infection,and study group added bronchoscopic alveolar lavage treatment based on the treatment of control group,and the anti-infection treatment regimen was adjusted according to the results of BALF lymphocyte immunoassay.Clinical improvement rates,clinical symptom scores,serum inflammatory factors[procal-citonin(PCT),C-reactive protein(CRP),TNF-α],peripheral blood lymphocyte subsets(CD3+T,CD4+T,CD8+T)levels,treatment condition,in-hospital mortality and incidence of acute respiratory distress syndrome(ARDS)between the two groups were compared.Results:Clinical improvement rates were higher in study group(44.87%,58.97%,91.03%,97.44%)than that in control group(26.92%,42.31%,78.21%,88.46%)at 3 d,5 d,7 d and 10 d of treatment(P<0.05);serum PCT,CRP and TNF-α levels were lower in study group than that in control group after 10 d of treatment(P<0.05);peripheral blood CD3+T,CD4+T,CD8+T levels were higher in study group than that in control group after 10 d of treatment(P<0.05).CPIS and CURB-65 scores were lower in study group than that in control group after 10 d of treatment(P<0.05);duration of antimicrobial drug use,duration of mechanical ventilation,and length of hospital stay were shorter in study group than that in control group(P<0.05);in-hospital mortality rate(7.69%)and inci-dence of ARDS(5.13%)were lower in study group than that in control group(19.23%,15.38%)(P<0.05).Conclusion:Optimized anti-infection treatment plan based on BALF cytology examination combined with lymphocyte immunoassay can significantly improve clinical efficacy of critically ill patients with SAP,and help to promote recovery and improve the prognosis of patients.

Objective:To summarize the clinical characteristics of autoimmune glial fibrillary acidic protein astrocytopathy(GFAP-A),in order to improve the understanding of the disease among clinical physicians.Methods:A retrospective analysis was conducted on the clinical manifestations,imaging examination,laboratory examination of a patient with GFAP-A who was admitted to the First People's Hospital of Yunnan Province and diagnosed on May 11,2023.Results:A female patient with GFAP-A,aged 67 years,presented with meningoencephalomyelitis.The head MRI showed abnormalities,with extensive lesions in the bilateral basal ganglia,pons,and lateral paraventricular white matter areas,and enhancement of the lateral ventricular ependyma and pia mater.Long segment spinal cord abnormal signals were observed,and cerebrospinal fluid anti GFAP antibodies were positive.After hormone treatment,the prognosis was good,and there was no recurrence during follow-up.Conclusion:The main clinical manifestations of GFAP-A are acute or subacute onset meningitis and/or encephalitis,myelitis and meningitis.Detection of autoimmune antibodies in ce-rebrospinal fluid can provide a clear diagnosis,and timely treatment can improve prognosis.

Objective:To explore the evaluation and relationship of hepatitis B virus(HBV)surface antigen(HBsAg)in clinical laboratory in different detection systems,further scientifically and reasonably to explain the test results for serving clinical practices.Methods:During the period from June 2021 to July 2022,100 425 specimens of patients with screened,suspected and confirmed HBV infections were collected from the clinical departments(mainly infectious hepatology)of Nanyang Central Hospital.Detection methodology included quantitative(electrochemiluminescence and chemiluminescence),qualitative(gold standard),semi-quantita-tive(ELISA),and highly sensitive HBV-DNA(RT-PCR)methods,and then analyzed the strengths and weaknesses and closeness be-tween each methodology.The relationship between the two Roche HBsAg detection systems was analyzed by correlation analysis.The HBsAg efficacy analysis was validated using Cut/Off value setting and detection limit,which in turn analyzed the distribution of false-positive and false-negative reporting models.Results:Detection results for low-and medium-concentration HBsAg showed a correla-tion between the electrochemical luminescence semi-quantitative method and ELISA method.ELISA method still had advantages in terms of sensitivity and specificity when detecting HBsAg,and there were no significant differences compared to domestic and interna-tional HBsAg quantitative detection systems.Performance validation conducted in accordance with the CNAS-GL038 document showed that the minimum detection limit for HBsAg calculated using the ELISA method in this laboratory was 0.1 U/ml.When the ROC curve Cut/Off value was set to 0.105,the area under the curve was the largest(AUC=0.986).Based on Roche's semi-quantitative electroche-miluminescence detection and patient medical history,in the common reporting model for hepatitis B five-item detection using ELISA method,HBsAg false positives occurred most frequently when HBsAg was positive alone,and HBsAg occurred most frequently in the false positive range when the OD value was less than 0.5.In ELISA method for detecting HBsAg,as the OD value increased from 0.01 to 0.10,the number of false-negative results also increased.Roche Elecsys HBsAg Ⅱ Quant Ⅱ and Elecsys HBsAg Ⅱ testing systems exhibited good linearity under certain conditions,with a ratio of approximately 1/0.18.In Elecsys HBsAg Ⅱ Quant Ⅱ detection sys-tem,the test results of HBsAg samples diluted 400 times were highly consistent with the original test results with a coefficient of deter-mination R2=0.993 8.Conclusion:There was a certain relationship between various detection systems of HBsAg at a suitable concen-tration.The detection of HBsAg by ELISA can meet the needs of clinical detection.

Objective:To construct three photosensitive fusion proteins between protein G C3 domain and Avitag for site-spe-cific biotinylation of goat IgG.Methods:DNA sequences of protein G C3 domain with amber codon mutation and C-terminal containing 1,2,3 Avitag were obtained by chemical synthesis,respectively,and sub-cloned into pTXB1 plasmid to construct three fusion pro-tein expression vectors.Three plasmids and pEVOL-pBpF plasmid were co-transformed into competent bacterium E.coli BL21,respec-tively.Target fusion proteins CBpa-Avitag,CBpa-(Avitag)2 and CBpa-(Avitag)3 were prepared using genetic codon extension technology,and were purified by His Trap Chelating HP column,then biotinylated by Avitag/BirA system.Photo-conjugation efficiency,crosslink site and antigen-binding activity of photo-conjugates were investigated by SDS-PAGE,control experiment,and competitive ELISA.Results:Three expression vectors were successfully constructed,and three designed photoactivatable fusion proteins CBpa-Avitag,CBpa-(Avitag)2 and CBpa-(Avitag)3 were successfully expressed with molecular weights consistent with their theoretical values.Heavy-chain conjugation efficiencies of biotinylated fusion protein with goat IgG were 85%with Fc-specific conjugation,and conjugates maintained its antigen-binding activity after photo-conjugation.Conclusion:CBpa-Avitag,CBpa-(Avitag)2 and CBpa-(Avitag)3 fusion proteins are successfully prepared,and mediates by those photosensitive fusion proteins,biotins molecules are conjugated to Fc fragment of goat IgG,results site-specific biotinylation of goat IgG.

The effective integration of ideological and political education into medical courses is one of the most important aspects of medical teaching reform and a key approach to fostering students'moral character.Based on the Outcomes-Based Education(OBE)philosophy,this article focuses on the learning outcomes of Medical Immunology ideological and political education,integrating relevant ideological and political elements with modular teaching content to form a themed"Medical Immunology Ideological and Politi-cal"teaching goal with distinctive features of Medical Immunology.Through a blended learning model of online and offline teaching methods,students are guided to learn modular teaching content.This study creatively integrates Medical Immunology theory with ideo-logical and political elements,carries out"Medical Immunology Ideological and Political"through professional knowledge,and achieves the goal of subtle and deep-seated ideological and political education.Moreover,it effectively improves students'comprehensive learning and application abilities,enabling them to apply what they have learned.

Veterinary Immunology is a science that studies organization structure and biological function of animal immune system.It is a subject with strong theory and application,which is a very important basic course in undergraduate education of veteri-nary medicine.With rapid development of modern science and technology,people have a deeper understanding of immunology,and traditional teaching content and teaching mode can no longer meet teaching needs of modern veterinary undergraduates.This paper briefly analyzes problems existing in current teaching process of Veterinary Immunology,and puts forward relevant suggestions for teaching reform,aiming at improving teaching quality of undergraduate Veterinary Immunology and shouldering important mission of cultivating innovative veterinary talents for socialist modernization of our country.

Objective:To investigate whether systemic immune inflammation index(SII)-C-reactive protein/albumin(CRP/Alb)can predict postoperative survival in non-small cell lung cancer(NSCLC)patients.Methods:A total of 262 patients with NSCLC who underwent surgery in Hebei Provincial People's Hospital from January 2017 to November 2020 were included.SII and CRP/Alb expressions in patients after surgery were analyzed by K-M survival curve analysis,Cox multivariate risk regression analysis and ROC curve analysis.Results:Patients with high SII expression(>961.78)and high CRP/Alb expression(>0.025)had worse outcomes than patients with low SII and CRP/Alb expressions,respectively(P<0.001).SII-CRP/Alb score(P=0.032),age≥60 years(P=0.038)and T stage T2～T3(P<0.001)were significantly associated with shorter overall survival.AUC of SII-CRP/Alb scores were larger than SII and CRP/Alb scores,respectively.Conclusion:SII-CRP/Alb score can effectively evaluate postoperative survival status of patients with NSCLC,and SII-CRP/Alb score are independent predictors of postoperative overall survival in NSCLC patients.

Objective:To investigate whether kisspeptin can influence the maternal-fetal interface regulatory T cells(Treg),thereby participating in the pathogenesis of recurrent spontaneous abortion(RSA).Methods:Normal pregnancy(NP)and RSA mice models were established,where NP mice received a tail vein injection of PBS(NP-PBS group),and RSA mice received a tail vein in-jection of PBS(RSA-PBS group)and active fragment of kisspeptin KP10(RSA-KP10 group),observing embryo absorption rates.Im-munohistochemistry was employed to assess expressions of kisspeptin and Foxp3 in mice uterine tissues.Peripheral blood Treg cells were isolated and expanded through magnetic bead separation.Intervention with KP10 and KP234(kisspeptin receptor antagonist)was administered,and flow cytometry was used to detect levels of IL-10 and TGF-β1 secretion by Treg cells,as well as differences in proliferation and apoptosis.RNA-Seq transcriptomic sequencing was conducted on uterine tissues from RSA-PBS group and RSA-KP10 group of mice.Differentially expressed genes(DEGs)were subjected to GO,KEGG and GSEA enrichment analyses.Results:Embryo absorption rate in RSA mice was higher than that in NP mice,the embryo absorption rate was decreased after tail vein injec-tion of KP10.Expressions of kisspeptin and Foxp3 in uterus of RSA mice was lower than that in NP mice,while increased after injec-tion of KP10.Kisspeptin could modulate the secretion of IL-10 and TGF-β1 by Treg cells,influencing their proliferation without affect-ing apoptosis.Enrichment analysis results showed that DEGs were mainly enriched in reproductive structure development,IL-17,and TGF-β signaling pathways.Conclusion:Kisspeptin can influence both the quantity and function of Treg cells,offering a new theoreti-cal foundation for investigating the pathogenesis and treatment of RSA.