The MGF300-4L protein of African swine fever virus(ASFV),a virulence-associated fac-tor,degrades IKKβ through the chaperone-mediated autophagy and enhances the stability of IKBαto suppress the generation of IL-1β and TNF-α regulated by the NF-κB signaling pathway.To iden-tify the host proteins interacting with MGF300-4L,PK-15 cells were transfected with the eukary-otic plasmid expressing MGF300-4L and analyzed using immunoprecipitation-mass spectrometry(IP-MS)to identify the host proteins that interact with MGF300-4L.Additionally,gene ontology(GO)and KEGG pathway enrichment analyses were conducted.Furthermore,molecular docking a-nalysis,co-immunoprecipitation,and laser confocal microscopy assays were performed to validate the host proteins interacting with MGF300-4L.The IP-MS analysis identified 145 host proteins that potentially interact with MGF300-4L.Subsequent GO and KEGG pathway enrichment analy-ses revealed that these proteins are predominantly involved in metabolic,cellular,and innate immune responses.Through molecular docking prediction,co-immunoprecipitation assay,and laser confocal microscopy,we identified the interaction between MGF300-4L and STAT1.This study provides critical insights into the mechanisms underlying the interactions between MGF300-4L and the host proteins.

The MGF300-4L protein of African swine fever virus(ASFV),a virulence-associated fac-tor,degrades IKKβ through the chaperone-mediated autophagy and enhances the stability of IKBαto suppress the generation of IL-1β and TNF-α regulated by the NF-κB signaling pathway.To iden-tify the host proteins interacting with MGF300-4L,PK-15 cells were transfected with the eukary-otic plasmid expressing MGF300-4L and analyzed using immunoprecipitation-mass spectrometry(IP-MS)to identify the host proteins that interact with MGF300-4L.Additionally,gene ontology(GO)and KEGG pathway enrichment analyses were conducted.Furthermore,molecular docking a-nalysis,co-immunoprecipitation,and laser confocal microscopy assays were performed to validate the host proteins interacting with MGF300-4L.The IP-MS analysis identified 145 host proteins that potentially interact with MGF300-4L.Subsequent GO and KEGG pathway enrichment analy-ses revealed that these proteins are predominantly involved in metabolic,cellular,and innate immune responses.Through molecular docking prediction,co-immunoprecipitation assay,and laser confocal microscopy,we identified the interaction between MGF300-4L and STAT1.This study provides critical insights into the mechanisms underlying the interactions between MGF300-4L and the host proteins.

African swine fever(ASF)is a highly contagious and pathogenic disease affecting both domestic and wild pigs,which is caused by African swine fever virus(ASFV).In European epidem-ics,low-virulence strains of ASFV,which do not have hemadsorbing properties,have been identi-fied.Following the identification of highly virulent genotype Ⅱ ASFV strains in China in 2018,subsequently,low-virulence strains of genotype Ⅱ and genotype Ⅰ emerged.Recombination be-tween genotypes Ⅰ and Ⅱ has also led to the occurrence of high-virulence strains.This indicates a complex and diverse genetic evolution of ASFV during the epidemiological transmission,which po-ses significant challenges for vaccine development and disease surveillance.Here,we provide an o-verview of the novel epidemiological characteristics of ASFV,with a focus on genetic variations and pathogenic differences during the outbreaks of ASF.We also explore how ASFV genetic varia-tions impact immune escape and pathogenicity of the virus,and the challenges they pose for vac-cine development,disease diagnosis,and surveillance.The aim of this review is to enhance our un-derstanding of the genetic evolution and mutation mechanisms of ASFV,providing a theoretical basis for the development of vaccines and research on diagnostic technologies.

Objective To explore the establishment of a subcutaneously transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome.Methods Forty male C57BL/6 mice were randomly divided into 4 groups:NC group,QZXY group,Tumor group,and QZXY+Tumor group.They were categorized based on the modeling of Qi stagnation and blood stasis syndrome(7 days)combined with the modeling of subcutaneous transplantation of hepatocellular carcinoma tumor(20 days).Observations were conducted of the syndrome manifestations as well as the tumor size and weight of the mice after modeling.Results(1)Body weight:on the 7th day of modeling,the weights of the QZXY group and QZXY+Tumor group were significantly lower than that of the NC group(P＜0.05).(2)Body temperature:on the 7th day of modeling,body temperature significantly decreased in the QZXY group(P＜0.05),while it increased in the Tumor group(P＜0.05)compared with the NC group.On the 27th day of modeling,the temperature of the QZXY+Tumor group was significantly lower than that of the NC group(P＜0.05).(3)Syndrome manifestations:according to the syndrome scoring table,mice in both the QZXY group and QZXY+Tumor group exhibited Qi stagnation and blood stasis syndrome on the 7th day of modeling(P＜0.05).As modeling time extended,the score of mice in the Tumor group increased with the formation of the tumor,and the score of mice in the QZXY+Tumor group was significantly higher than that of the other three groups(P＜0.05).(4)Claw petechiae:the number of claw petechiae significantly increased in all three groups of modeled mice compared with the NC group(P＜0.05),with the QZXY+Tumor group showing the highest number.(5)Claw r value:the r value of the claw was significantly lower in all three groups of modeled mice than that in the NC group(P＜0.05).Additionally,the r value of the claw in the QZXY+Tumor group was consistently lower than that of the other three groups.(6)Open field activity:the vertical and horizontal activity of mice in the QZXY+Tumor group decreased significantly compared with that of the NC group(P＜0.05).(7)Coagulation indexes:APTT,TT,and FIB were significantly increased in the QZXY+Tumor group(P＜0.05 or P＜0.01)compared with those in the NC group.(8)Tumor size and weight:compared with the Tumor group,the QZXY+Tumor group showed significantly increased tumor size and weight(P＜0.05).Conclusions This study successfully established a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome.The findings indicated that Qi stagnation and blood statsis syndrome may occur during the course of live cancer.Besides,the causes inducing the Qi stagnation and blood stasis syndrome will further accelerate the progression of liver cancer.

Objective To investigate the molecular mechanism of Lizhong Wan in treatment of chronic atrophic gastritis (CAG) using network pharmacology and in vitro experimental method. Methods The active ingredients of Lizhong Wan were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the related targets were obtained from PubChem and Swiss Target Prediction databases. The CAG-related targets were obtained from GeneCards,OMIM and DisGeNET databases. The drug-disease common targets of Lizhong Wan and CAG were obtained by Venn tool. The interaction network of common targets of Lizhong Wan and CAG was constructed with the help of STRING database,and the drug-disease-target-pathway network was constructed with the help of Cytoscape software. Gene function and pathway enrichment analyses were performed on the common targets. The molecular docking and binding ability of the active ingredients of the drug and the key targets were predicted by using Moe software. The mice of spleen and stomach deficiency cold syndrome CAG was established. The effects of Lizhong Wan on the morphological changes of gastric tissue,apoptosis of gastric mucosa cells and the expression of messenger RNA (mRNA) and protein of the key target of CAG were observed. Results A total of 57 active ingredients of Lizhong Wan were screened,including arachidonic acid,ginsenoside Rg5,gomisin B,aposiopolamine and ginsenoside Rh2. 869 active targets of Lizhong Wan were obtained. CAG and Lizhong Wan had 47 common targets,the key common targets including tumor protein P53 (TP53),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),epidermal growth factor receptor (EGFR),B-cell lymphoma 2 (Bcl-2),etc.. A total of 135 pathways were obtained by enrichment analysis,mainly including tumor pathogenesis,proteoglycan in tumor,cholinergic synapse,phosphoinositide 3-kinase/protein kinase B signaling pathway,etc.. Molecular docking results showed that TP53,IL-6,TNF-α,EGFR,Bcl-2 had good binding activity with gomisin B and ginsenoside Rh2. Meanwhile,in vitro experimental found that the scores of spleen and stomach deficiency cold syndrome in model group were significantly higher than those in blank group. Lizhong Wan could improve the pathological changes of CAG mice,could significantly reduce the apoptosis of gastric mucosa cells,could significantly reduce the expression of mRNA and protein of TP53,IL-6,TNF-α,EGFR,Bcl-2. Conclusion The effect of Lizhong Wan in treating CAG with spleen and stomach deficiency cold syndrome may be related to regulating the expression of TP53,IL-6,TNF-α,EGFR and Bcl-2,alleviating inflammation and reducing apoptosis of gastric mucosa cells.

Objective To explore the establishment of a subcutaneously transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome.Methods Forty male C57BL/6 mice were randomly divided into 4 groups:NC group,QZXY group,Tumor group,and QZXY+Tumor group.They were categorized based on the modeling of Qi stagnation and blood stasis syndrome(7 days)combined with the modeling of subcutaneous transplantation of hepatocellular carcinoma tumor(20 days).Observations were conducted of the syndrome manifestations as well as the tumor size and weight of the mice after modeling.Results(1)Body weight:on the 7th day of modeling,the weights of the QZXY group and QZXY+Tumor group were significantly lower than that of the NC group(P＜0.05).(2)Body temperature:on the 7th day of modeling,body temperature significantly decreased in the QZXY group(P＜0.05),while it increased in the Tumor group(P＜0.05)compared with the NC group.On the 27th day of modeling,the temperature of the QZXY+Tumor group was significantly lower than that of the NC group(P＜0.05).(3)Syndrome manifestations:according to the syndrome scoring table,mice in both the QZXY group and QZXY+Tumor group exhibited Qi stagnation and blood stasis syndrome on the 7th day of modeling(P＜0.05).As modeling time extended,the score of mice in the Tumor group increased with the formation of the tumor,and the score of mice in the QZXY+Tumor group was significantly higher than that of the other three groups(P＜0.05).(4)Claw petechiae:the number of claw petechiae significantly increased in all three groups of modeled mice compared with the NC group(P＜0.05),with the QZXY+Tumor group showing the highest number.(5)Claw r value:the r value of the claw was significantly lower in all three groups of modeled mice than that in the NC group(P＜0.05).Additionally,the r value of the claw in the QZXY+Tumor group was consistently lower than that of the other three groups.(6)Open field activity:the vertical and horizontal activity of mice in the QZXY+Tumor group decreased significantly compared with that of the NC group(P＜0.05).(7)Coagulation indexes:APTT,TT,and FIB were significantly increased in the QZXY+Tumor group(P＜0.05 or P＜0.01)compared with those in the NC group.(8)Tumor size and weight:compared with the Tumor group,the QZXY+Tumor group showed significantly increased tumor size and weight(P＜0.05).Conclusions This study successfully established a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome.The findings indicated that Qi stagnation and blood statsis syndrome may occur during the course of live cancer.Besides,the causes inducing the Qi stagnation and blood stasis syndrome will further accelerate the progression of liver cancer.

Objective To investigate the molecular mechanism of Lizhong Wan in treatment of chronic atrophic gastritis (CAG) using network pharmacology and in vitro experimental method. Methods The active ingredients of Lizhong Wan were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the related targets were obtained from PubChem and Swiss Target Prediction databases. The CAG-related targets were obtained from GeneCards,OMIM and DisGeNET databases. The drug-disease common targets of Lizhong Wan and CAG were obtained by Venn tool. The interaction network of common targets of Lizhong Wan and CAG was constructed with the help of STRING database,and the drug-disease-target-pathway network was constructed with the help of Cytoscape software. Gene function and pathway enrichment analyses were performed on the common targets. The molecular docking and binding ability of the active ingredients of the drug and the key targets were predicted by using Moe software. The mice of spleen and stomach deficiency cold syndrome CAG was established. The effects of Lizhong Wan on the morphological changes of gastric tissue,apoptosis of gastric mucosa cells and the expression of messenger RNA (mRNA) and protein of the key target of CAG were observed. Results A total of 57 active ingredients of Lizhong Wan were screened,including arachidonic acid,ginsenoside Rg5,gomisin B,aposiopolamine and ginsenoside Rh2. 869 active targets of Lizhong Wan were obtained. CAG and Lizhong Wan had 47 common targets,the key common targets including tumor protein P53 (TP53),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),epidermal growth factor receptor (EGFR),B-cell lymphoma 2 (Bcl-2),etc.. A total of 135 pathways were obtained by enrichment analysis,mainly including tumor pathogenesis,proteoglycan in tumor,cholinergic synapse,phosphoinositide 3-kinase/protein kinase B signaling pathway,etc.. Molecular docking results showed that TP53,IL-6,TNF-α,EGFR,Bcl-2 had good binding activity with gomisin B and ginsenoside Rh2. Meanwhile,in vitro experimental found that the scores of spleen and stomach deficiency cold syndrome in model group were significantly higher than those in blank group. Lizhong Wan could improve the pathological changes of CAG mice,could significantly reduce the apoptosis of gastric mucosa cells,could significantly reduce the expression of mRNA and protein of TP53,IL-6,TNF-α,EGFR,Bcl-2. Conclusion The effect of Lizhong Wan in treating CAG with spleen and stomach deficiency cold syndrome may be related to regulating the expression of TP53,IL-6,TNF-α,EGFR and Bcl-2,alleviating inflammation and reducing apoptosis of gastric mucosa cells.

Cell-mediated immune response is an important part of machinery in maintaining the body's homeostasis. After the innate immune system selectively activates the adaptive immune system, the cell-mediated immunity exerts its killing and clearance functions. Therefore, evaluating the level of cell-mediated immune response is crucial in the diagnosis and treatment of cancer, monitoring the immune status after organ transplantation, diagnosing and preventing viral diseases, and evaluating the effectiveness of vaccines and other areas. From the initial overall assessment of the immune effects in vivo to the precise detection of the number and function of multiple immune cells, the evaluation methods of cell-mediated immune response have greatly advanced. However, cell-mediated immune response involves multiple levels in the body, and it's difficult to choose the numerous detection methods available. The article systematically compares the evaluation methods of cell-mediated immune response at four different levels: the organism, the tissue and organ, the immune cells and the immune molecules, with the aim to facilitate the applications of related technologies.
Humans ; Immunity, Cellular ; Neoplasms/therapy* ; Immunity, Innate

Objective:To study the changes of β-lactam resistance of Haemophilus influenzae (Hi) strain isolated from neonatal lower respiratory tract and the molecular mechanism of β-lactam resistance.Methods:Nineteen Hi strains isolated from neonatal lower respiratory tract infection in the previous multicenter prospective epidemiological study were re-identified, and the P6, fucK and Cap genes were detected by PCR.The minimum inhibitory concentration(MIC) of ampicillin, amoxicillin clavulanic acid and cefuroxime were detected by microdilution method, and tem-1 gene, rob-1 gene and ftsI gene were sequenced and analyzed.Results:(1) Nineteen strains of Hi were confirmed to be capsule-free type by P6 gene, fucK gene and cap gene, which was non-typeable Haemophilus influenzae(NTHi). (2)Compared with 2003-2004, the MIC values of ampicillin, amoxicillin clavulanic acid and cefuroxime of NTHi isolated from the lower respiratory tract of the newborn from 2013-2014 were significantly higher( P<0.05). (3)The rates of β-lactamase producing strains during 2003-2004 and 2013-2014 were 33.33% (3/9) and 30.00% (3/10), respectively.There was no significant difference between them during 10 years ( P>0.05). The detection of the β-lactamase gene showed that the β-lactamase of the all six strains were of the tem-1 type, and the rob-1 type was not detected.(4)Only one gBLNAR strain ( n=9) was found during 2003~2004, and gBLNAR 1, gBLNAI 3, gBLPAR 3, gBLPACR 1 ( n=10)appeared during 2013~2014.(5)There were 11 amino acid substitution patterns in ftsI gene during 2013~2014, but only five amino acid substitution patterns in 2003~2004.The mutation rate of the S357N, S385T, N526K and T532S of ftsI gene significantly increased during the past ten years ( P<0.05). One strain of gBLNAR/gBLNACR resistant to ampicillin, amoxicillin clavulanic acid and cefuroxime isolated in 2014 showed D350N, S357N, M377I, S385T, L389F, A502T and N526K variation at the same time. Conclusion:Neonatal patients with lower respiratory tract NTHi infection may rapidly face the severe challenge of multiple drug resistance of β-lactam antibiotics.

Objective Broth dilution method was used as a reference method to observe the capability of Kirby-Bauer disc diffusion assay(K-B)for correcting automated ampicillin susceptibility detection of He-mophilus influenzae(HI).Methods A total of 228 HI strains isolated were collected,broth dilution assay,K-B and automated microdilution broth test(ATB)were used to determine the susceptibility of HI to ampicillin. Analyze the essential agreements of the three methods and the correction of K-B to the errors of A TB. Results The essential agreement of K-B or ATB with broth dilution method were 77.19%,70.18% respec-tively,combination of K-B and ATB could make the essential agreement increase up to 86.0%,which was sig-nificantly higher than ATB(χ2=16.600,P=0.000).Major error of ATB(42.0%)was higher than that of K-B(10.0%)(χ2=13.306,P=0.001),but very major error and minor error showed no significant difference be-tween the two methods(χ2=1.208,P=0.272;χ2=1.182,P=0.227),meanwhile,76.19% of major error of ATB could be corrected by K-B.For the very major error of ATB,53.57% could be corrected by K-B.Howev-er,the corrective capability of K-B to minor error of ATB was relative low.Conclusion K-B test could correct some errors generated by ATB.For the β-lactamase negative strains which were judged as ampicillin resistance by A TB,K-B test should be used to correct the errors by ATB.Moreover,it is necessary to apply K-B to confirm am-picillin sensitivity of the β-lactamase positive strains which were judged as ampicillin susceptible by ATB.