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 effects of picroside Ⅱ(PⅡ)on proliferation,apoptosis and immune escape of lung cancer cells by regulating C-C motif chemokine ligand 2(CCL2)/C-C motif chemokine receptor 2(CCR2)signaling axis.Methods:Human lung cancer cells NCI-H292 were cultured and treated with 0,5,10,20,40 and 80 μmol/L PⅡ,MTT method was applied to detect cell viability.Experiment was separated into control group,low,medium and high concentrations PⅡ groups(PⅡ-L,PⅡ-M,PⅡ-H,10,20 and 40 μmol/L PⅡ),high concentration PⅡ+CCL2 overexpression negative control group(PⅡ-H+pcDNA-NC,40 μmol/L PⅡ+pcDNA-NC)and high concentration PⅡ+CCL2 overexpression group(PⅡ-H+CCL2,40 μmol/L PⅡ+pcDNA-CCL2).EdU method was applied to measure cell proliferation;flow cytometry was applied to measure cell apoptosis;immunoblotting was applied to determine expressions of CCL2,CCR2,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax).Lung can-cer cells in each group were co-cultured with CD8+T cells,Trypan blue staining was applied to measure CD8+T cell viability;ELISA was applied to determine levels of programmed death receptor-ligand 1(PD-L1),IL-10,IFN-γ and TGF-β.Results:Compared with 0 μmol/L,cell viability treated with 10,20,40 and 80 μmol/L PⅡ were significantly reduced(P<0.05),and 10,20 and 40 μmol/L PⅡ were selected for subsequent experiments.Compared with control group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while apoptosis rate and expression of Bax were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while apoptosis rate and expression of Bax were de-creased significantly(P<0.05).After co-culturing with CD8+T cells,compared with control group,levels of IL-10,TGF-β and PD-L1 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while CD8+T cell viability and level of IFN-γ were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,levels of IL-10,TGF-β and PD-L1 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while CD8+T cell viability and level of IFN-γ were reduced significantly(P<0.05).Conclu-sion:PⅡ may inhibit proliferation and immune escape of lung cancer cells,and promote cell apoptosis by inhibiting CCL2-CCR2 sig-naling axis.

Objective To investigate the effects of asiaticoside(AS)on epithelial-mesenchymal transition(EMT)and radiotherapy sensitivity of esophageal cancer(EC)cells by its mechanism of regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods EC9706 cells were subjected to different concentrations of AS or different doses of radiation.Methyl thiazolyl tetrazolium method was used to detect cell proliferation and calculate the half-maximal inhibitory con-centration.EC9706 cells were divided into a control group,radiology group(X-ray irradiation),AS group,combined group(AS+X-ray irradiation),and activator group(AS+X-ray irradiation+HIF-1α/VEGF pathway activator dimethyloxallyl glycine).Plate cloning experi-ments were conducted to detect sensitivity,and Transwell assays were used to detect cell migration and invasion.Flow cytometry helped detect apoptosis,and real-time quantitative polymerase chain reaction detected the expression of HIF-1α and VEGF mRNA.Western blotting method was used to detect the expression of matrix metalloproteinase-2(MMP-2),vimentin,E-cadherin,Bcl-2 associated X protein(Bax),HIF-1α,and VEGF proteins.Results With the increase of AS concentration and radiation dose,the cell viability of EC9706 cells gradually decreased;compared with the control group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1α and VEGF mRNA;and the expression of MMP-2,vimentin,HIF-1α,and VEGF in the radiology group and AS group were reduced;further,the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).Compared with the radiology group and AS group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1αand VEGF mRNA;and the expressions of MMP-2,vimentin,HIF-1α,and VEGF in the combined group were reduced;the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).In comparison with the combined group,the changes in the above indicators in the activator group were reversed(P＜0.05).Conclusion AS may inhibit EMT by inhibiting the HIF-1α/VEGF signaling pathway,thus enhancing the radiotherapy sensitivity of EC cells.

Background and purpose:The aberrant activation of pyruvate dehydrogenase kinase 1(PDK1)drives tumor microenvironment remodeling and metastasis through mediating the Warburg effect.As a critical tumor-suppressive phosphatase,phosphatase and tensin homolog deleted on chromoseme ten(PTEN)activates PDK1 via loss of expression to induce aerobic glycolysis and accelerate tumor progression.The molecular interplay between PDK1 and PTEN in kidney renal clear cell carcinoma(KIRC)urgently requires systematic elucidation.This study aimed to clarify how PTEN regulates PDK1 to inhibit malignant phenotypes in KIRC.Methods:Bioinformatics analysis was conducted to compare PTEN and PDK1 expression levels as well as their prognostic correlations in the Cancer Genome Atlas(TCGA)-KIRC datasets.KIRC cell models was established by either silencing PDK1 or enhancing its expression,subsequently evaluating their malignancy characteristics through cell counting kit-8(CCK-8)proliferation,colony formation,cell migration,and invasion assays.To validate the regulatory interactions,we used PDK1-overexpressing cells treated with a PTEN-specific inhibitor.Western blot was used to dectect the protein expression.Results:The TCGA-KIRC analysis found significantly higher mRNA levels of PTEN and PDK1 in tumor tissues compared to normal controls(P＜0.05),yet this high expression was associated with improved overall survival(P＜0.01).Besides,a strong positive correlation was observed between PTEN and PDK1 expressions(r=0.52,P＜0.001).Functional assays demonstrated that PDK1 knockdown markedly promoted cell proliferation,migration,and invasion,whereas PDK1 overexpression exhibited opposing effects.Mechanistically,inhibiting PTEN worsened malignant behaviors(P＜0.01),however,these effects were reversed by overexpressing PDK1.Conclusion:This study presents the first evidence of the dual tumor-suppressive function of the PTEN-PDK1 biological axis in renal cancer,which supports the development of precision treatment strategies based on novel targets.

Objective:To investigate effects of picroside Ⅱ(PⅡ)on proliferation,apoptosis and immune escape of lung cancer cells by regulating C-C motif chemokine ligand 2(CCL2)/C-C motif chemokine receptor 2(CCR2)signaling axis.Methods:Human lung cancer cells NCI-H292 were cultured and treated with 0,5,10,20,40 and 80 μmol/L PⅡ,MTT method was applied to detect cell viability.Experiment was separated into control group,low,medium and high concentrations PⅡ groups(PⅡ-L,PⅡ-M,PⅡ-H,10,20 and 40 μmol/L PⅡ),high concentration PⅡ+CCL2 overexpression negative control group(PⅡ-H+pcDNA-NC,40 μmol/L PⅡ+pcDNA-NC)and high concentration PⅡ+CCL2 overexpression group(PⅡ-H+CCL2,40 μmol/L PⅡ+pcDNA-CCL2).EdU method was applied to measure cell proliferation;flow cytometry was applied to measure cell apoptosis;immunoblotting was applied to determine expressions of CCL2,CCR2,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax).Lung can-cer cells in each group were co-cultured with CD8+T cells,Trypan blue staining was applied to measure CD8+T cell viability;ELISA was applied to determine levels of programmed death receptor-ligand 1(PD-L1),IL-10,IFN-γ and TGF-β.Results:Compared with 0 μmol/L,cell viability treated with 10,20,40 and 80 μmol/L PⅡ were significantly reduced(P<0.05),and 10,20 and 40 μmol/L PⅡ were selected for subsequent experiments.Compared with control group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while apoptosis rate and expression of Bax were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while apoptosis rate and expression of Bax were de-creased significantly(P<0.05).After co-culturing with CD8+T cells,compared with control group,levels of IL-10,TGF-β and PD-L1 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while CD8+T cell viability and level of IFN-γ were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,levels of IL-10,TGF-β and PD-L1 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while CD8+T cell viability and level of IFN-γ were reduced significantly(P<0.05).Conclu-sion:PⅡ may inhibit proliferation and immune escape of lung cancer cells,and promote cell apoptosis by inhibiting CCL2-CCR2 sig-naling axis.

Objective To investigate the effects of asiaticoside(AS)on epithelial-mesenchymal transition(EMT)and radiotherapy sensitivity of esophageal cancer(EC)cells by its mechanism of regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods EC9706 cells were subjected to different concentrations of AS or different doses of radiation.Methyl thiazolyl tetrazolium method was used to detect cell proliferation and calculate the half-maximal inhibitory con-centration.EC9706 cells were divided into a control group,radiology group(X-ray irradiation),AS group,combined group(AS+X-ray irradiation),and activator group(AS+X-ray irradiation+HIF-1α/VEGF pathway activator dimethyloxallyl glycine).Plate cloning experi-ments were conducted to detect sensitivity,and Transwell assays were used to detect cell migration and invasion.Flow cytometry helped detect apoptosis,and real-time quantitative polymerase chain reaction detected the expression of HIF-1α and VEGF mRNA.Western blotting method was used to detect the expression of matrix metalloproteinase-2(MMP-2),vimentin,E-cadherin,Bcl-2 associated X protein(Bax),HIF-1α,and VEGF proteins.Results With the increase of AS concentration and radiation dose,the cell viability of EC9706 cells gradually decreased;compared with the control group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1α and VEGF mRNA;and the expression of MMP-2,vimentin,HIF-1α,and VEGF in the radiology group and AS group were reduced;further,the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).Compared with the radiology group and AS group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1αand VEGF mRNA;and the expressions of MMP-2,vimentin,HIF-1α,and VEGF in the combined group were reduced;the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).In comparison with the combined group,the changes in the above indicators in the activator group were reversed(P＜0.05).Conclusion AS may inhibit EMT by inhibiting the HIF-1α/VEGF signaling pathway,thus enhancing the radiotherapy sensitivity of EC cells.

Background and purpose:The aberrant activation of pyruvate dehydrogenase kinase 1(PDK1)drives tumor microenvironment remodeling and metastasis through mediating the Warburg effect.As a critical tumor-suppressive phosphatase,phosphatase and tensin homolog deleted on chromoseme ten(PTEN)activates PDK1 via loss of expression to induce aerobic glycolysis and accelerate tumor progression.The molecular interplay between PDK1 and PTEN in kidney renal clear cell carcinoma(KIRC)urgently requires systematic elucidation.This study aimed to clarify how PTEN regulates PDK1 to inhibit malignant phenotypes in KIRC.Methods:Bioinformatics analysis was conducted to compare PTEN and PDK1 expression levels as well as their prognostic correlations in the Cancer Genome Atlas(TCGA)-KIRC datasets.KIRC cell models was established by either silencing PDK1 or enhancing its expression,subsequently evaluating their malignancy characteristics through cell counting kit-8(CCK-8)proliferation,colony formation,cell migration,and invasion assays.To validate the regulatory interactions,we used PDK1-overexpressing cells treated with a PTEN-specific inhibitor.Western blot was used to dectect the protein expression.Results:The TCGA-KIRC analysis found significantly higher mRNA levels of PTEN and PDK1 in tumor tissues compared to normal controls(P＜0.05),yet this high expression was associated with improved overall survival(P＜0.01).Besides,a strong positive correlation was observed between PTEN and PDK1 expressions(r=0.52,P＜0.001).Functional assays demonstrated that PDK1 knockdown markedly promoted cell proliferation,migration,and invasion,whereas PDK1 overexpression exhibited opposing effects.Mechanistically,inhibiting PTEN worsened malignant behaviors(P＜0.01),however,these effects were reversed by overexpressing PDK1.Conclusion:This study presents the first evidence of the dual tumor-suppressive function of the PTEN-PDK1 biological axis in renal cancer,which supports the development of precision treatment strategies based on novel targets.

Objective:To establish a carbohydrate metabolism-related genes(CRGs)prognostic model for clear cell renal cell carcinoma(ccRCC)and investigate its clinical value.Methods:ccRCC mRNA expression data were sourced from The Cancer Genome Atlas(TCGA)data-base.CRGs were retrieved from the MSigDB and KEGG databases.A prognostic model based on CRGs was constructed using the LASSO lin-ear regression model,and the risk score(RS)was calculated.Patients were assigned into high-and low-risk groups according to the median RS.Differences in survival,immune infiltration,mutation,and immune response between the two groups were analyzed using Kaplan-Meier curves and bioinformatics methods.Constructing a nomogram based on the RS and clinical features and validating its accuracy of prognostic predictions.The expression of CRGs in the ccRCC samples was detected using RT-qPCR.Results:A total of eight key genes were utilized to construct a prognostic risk model for ccRCC.Survival analysis revealed that patients in the low-risk group had a better prognosis(P<0.001).Bioinformatics analysis showed that the RS correlated with immune cell infiltration,mutation,and immune responses.The nomogram based on the RS and clinical features demonstrated a strong predictive ability for prognosis.In vitro experiments confirmed notable differences in the expression of the eight CRGs between ccRCC and adjacent non-malignant tissues.Conclusions:A prognostic model based on CRGs can effectively predict the prognosis of patients with ccRCC.

OBJECTIVE To explore the intestinal absorption characteristics of saikosaponins. METHODS Based on everted intestinal sac model， using accumulative absorption amount （Q） and absorption rate constant （Ka） as indexes， UHPLC-MS/MS technique as a method， the absorption of saikosaponin A， B2， C， D and F from total saponins of Bupleurum chinense （8 g/mL， by crude drug） in the duodenum， jejunum and ileum was detected. RESULTS The correlation coefficients （r） of the regression equations for the absorption of saikosaponins A， B2， C and F in the duodenum， jejunum and ileum were all higher than 0.95， while the r of saikosaponin D in the above intestinal segments was lower than 0.95； compared with the absorption of the same composition in the duodenum， the Q and Ka of saikosaponin A and C circulating in jejunum and ileum for 120 min， as well as the Q and Ka of saikosaponin F circulating in the ileum for 120 min were significantly decreased （P＜0.05）. CONCLUSIONS Saikosaponin A and the other 4 saikosaponins are all absorbed in the duodenum， jejunum and ileum； among them， saikosaponin A， B2， C and F are linearly absorbed， which conforms to the zero-order absorption characteristics， but saikosaponin D shows non- linear absorption.

Objective To evaluate the manifestations and diagnostic value of CT enterography (CTE) in primary intestinal T-cell lymphoma (PITCL).Methods Eighteen patients with PITCL confirmed by pathology were reviewed retrospec tively.The characteristics of lesion site,amount of foci,pattern and degree of contrast enhancement,lymphadenopathy,involvement of other organs and complications were recorded.Results In all of the 18 patients with PITCL,multiple lesions were seen in 13 cases (13/18,72.22％),and solitary involvement was seen in 5 cases (5/18,27.78％).Twelve ca ses were located at jejunum/ileum,3 of them were also involved in the colon.Five cases were located only in the colon,and 1 in the duodenum.Six cases were complicated with intestinal perforation.The patients were categorized into 6 types according to the CT manifestation:infiltration type (n=7),diffuse jejunum mucosa ileum metaplasia type (n =3),luminal aneurismal dilatation type (n =3),polypoid mass type (n =2),mesentery type (n=1),mixed type (n =2).Conclusion CTE can clearly display the imaging of PITCL and it has high value for the diagnosis of PITCL.