HDAC7, a member of class IIa HDACs, plays a pivotal regulatory role in tumor, immune, fibrosis, and angiogenesis, rendering it a potential therapeutic target. Nevertheless, due to the high similarity in the enzyme active sites of class IIa HDACs, inhibitors encounter challenges in discerning differences among them. Furthermore, the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes, leading to a limited impact of enzymatic inhibitors on their function. In this study, proteolysis targeting chimera (PROTAC) technology was employed to develop HDAC7 drugs. We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma (DLBCL) and acute myeloid leukemia (AML) cells. Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7, thereby exerting proliferative inhibition in DLBCL. Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies, particularly in DLBCL and AML.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

Objective To observe the value of parallel reverse enhance attention network module based on Kolmogorov-Arnold networks(KAN-PrdaModule)for segmenting polyps showed on colonoscopy images.Methods Nine hundred colonoscopy images in Kvasir-SEG dataset and 550 colonoscopy images in CVC-ClinicDB dataset were selected as training set(n=1 450),while 196 colonoscopy images in ETIS dataset,62 in CVC-ClinicDB dataset,380 in CVC-ColonDB dataset and 100 in Kvasir-SEG dataset were enrolled as test set(n=738).KAN-PrdaModule was proposed through improving U-Net,which improved detection accuracy through multi-scale feature fusion,and the value of KAN-PrdaModule for segmenting polyps showed on colonoscopy images was analyzed according to mean Dice similarity coefficient(mDSC),mean intersection over union(mIoU),weighted metric(Fωβ)and structural metric(Sα),and compared with U-Net,U-Net++,stochastic frontier analysis(SFA)and PraNet models.Results Among the above 5 models,the performance of SFA model for segmenting polyps on colonoscopy images was poor,with blurry edges of polyps and some ones were missed.U-Net and U-Net++models had decent performance,which could roughly identify polyps.PraNet model performed well,and the segmented edges of polyps were clear.KAN-PrdaModule had the best performance,showed high similarity to the true value images,with the best overall mDSC,mIoU,Fωβand Sα.Conclusion KAN-PrdaModule could effectively segment polyps showed on colonoscopy images,with segmenting effect better than U-Net,U-Net++,SFA and PraNet models.

Objective　To investigate the effect of andrographolide on enterovirus 71 (EV71) and coxsackievirus A16 (CA16) in vitro. Methods Cytopathic CPE assay and thiazolyl blue tetrazolium bromide (MTT) assay were used to determine the maximum non-toxic dose of andrographolide on RD cells and the inhibitory effect of andrographolide on EV71 and CA16 infection in vitro. The effects of andrographolide on the virus VP1 gene and cellular inflammatory factors (IL-1β, IL-6, and TNF-α) at gene levels were detected by quantitative fluorescence PCR. Results MTT results showed that the maximum non-toxic dose of andrographolide on RD cells was 78.00 μg/mL. Upon infection with hand-foot-mouth disease viruses, cells treated with andrographolide solutions at concentrations of 78.00, 39.00, 19.50, 9.75, 4.88, and 2.44 μg/mL showed increased survival rates to (82.41±1.76)%, (79.54±2.91)%, (81.02±1.99)%, (71.81±2.26)%, (52.87±1.51)%, and (50.41±0.93)% for EV71 and (81.00±0.64)%, (79.72±1.38)%, (61.59±3.47)%, (53.37±0.53)%, (52.41±1.37)%, and (43.69±0.40)% for CA16, respectively, indicating a significant reduction in the cytopathic effect caused by EV71 and CA16. When infected with hand-foot-mouth viruses and treated with andrographolide solutions at concentrations of 78.00, 39.00, 19.50, and 9.75 μg/mL, the expression levels of VP1 in EV71 and CA16, along with pro-inflammatory factors IL-1β, IL-6, and TNF-α in RD cells, were significantly lower compared to the virus control group. These results indicated that after infection with hand-foot-mouth viruses, treatment with andrographolide solution significantly inhibits the expression of the VP1 gene and reduces the mRNA levels of IL-1β, IL-6, and TNF-α. Conclusions Andrographolide exhibits obvious in vitro antiviral effects against EV71 and, potentially through the inhibition of the expression of pro-inflammatory factors IL-1β, TNF-α, and IL-6, thereby exerting its antiviral effects.

Guided by ZHANG Jiebin's "water-fire of life gate theory"， this paper explored the etiology， pathoge-nesis， and syndrome differentiation-based treatment approach for esophageal cancer. It is proposed that the core pathogenesis of esophageal cancer， derived from the imbalanced state of water and fire in life gate， can be summarized into three manifestations. Firstly， decline of life gate fire leads to failure of qi transformation and accumulation of phlegm and blood stasis； secondly， fire fails to warm the earth resulting in dysfunctional ascending and descending， and qi stagnation and blood stasis； thirdly， deficiency of life gate water contributs to proliferation of cancerous toxins and physical deterioration. In terms of treatment， guided by ZHANG's treatment principles for dysphagia， three treatment methods are proposed， warming and tonifying kidney yang to promote qi transformation through steaming action， warming and nourishing the spleen and stomach to restore central pivot movement， and nourishing kidney water to replenish essence and blood， then support bodily nourishment. Clinical case examples are provided to illustrate the application of these methods.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

Objective To observe the value of parallel reverse enhance attention network module based on Kolmogorov-Arnold networks(KAN-PrdaModule)for segmenting polyps showed on colonoscopy images.Methods Nine hundred colonoscopy images in Kvasir-SEG dataset and 550 colonoscopy images in CVC-ClinicDB dataset were selected as training set(n=1 450),while 196 colonoscopy images in ETIS dataset,62 in CVC-ClinicDB dataset,380 in CVC-ColonDB dataset and 100 in Kvasir-SEG dataset were enrolled as test set(n=738).KAN-PrdaModule was proposed through improving U-Net,which improved detection accuracy through multi-scale feature fusion,and the value of KAN-PrdaModule for segmenting polyps showed on colonoscopy images was analyzed according to mean Dice similarity coefficient(mDSC),mean intersection over union(mIoU),weighted metric(Fωβ)and structural metric(Sα),and compared with U-Net,U-Net++,stochastic frontier analysis(SFA)and PraNet models.Results Among the above 5 models,the performance of SFA model for segmenting polyps on colonoscopy images was poor,with blurry edges of polyps and some ones were missed.U-Net and U-Net++models had decent performance,which could roughly identify polyps.PraNet model performed well,and the segmented edges of polyps were clear.KAN-PrdaModule had the best performance,showed high similarity to the true value images,with the best overall mDSC,mIoU,Fωβand Sα.Conclusion KAN-PrdaModule could effectively segment polyps showed on colonoscopy images,with segmenting effect better than U-Net,U-Net++,SFA and PraNet models.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.