Objective:To investigate the efficacy and safety of bendamustine combined with anti-CD20 monoclonal antibody in the first-line treatment of older patients with indolent B-cell non-Hodgkin lymphoma (B-iNHL) .Methods:The clinical data of 159 patients with B-iNHL enrolled in 16 hospitals from Jiangsu Cooperative Lymphoma Group from December 1, 2019, to April 20, 2024, were analyzed for regimen efficacy and safety. Bendamustine plus rituximab (BR) and bendamustine plus obinutuzumab (BG) were administered to 139 (87.4% ) and 20 (12.6% ) patients, respectively.Results:Among the 159 patients, 101 (63.5% ) were male and 58 (36.5% ) were female, with a median age of 69 years (range: 60–84). Efficacy could be assessed in 138 (86.8% ) patients. The efficacy assessment demonstrated that the overall response rate was 92.0% with complete and partial remissions in 75 (54.3% ) and 52 (37.7% ) cases, respectively. With a median follow-up of 24 months (range: 4–64), the progression-free survival rate was (87.5 ± 3.0) % and the overall survival rate was (83.2 ± 3.3) %. Of the 27 patients who died, 6 (22.2% ) died due to disease progression. The mean applied dose of bendamustine per cycle was 73.0 (50.8–89.7) mg/m 2 per day, administered on days 1 and 2. Adverse events of grade 3 or higher were reported in 53 (33.3% ) patients, with infection (30 cases,18.9% ) and neutropenia (24 cases, 15.1% ) demonstrating the highest incidence. Conclusion:Bendamustine combined with anti-CD20 monoclonal antibody demonstrated good efficacy and is well-tolerated in the first-line treatment of elderly patients with B-iNHL.

Objective To investigate the expression and significance of 25-hydroxyvitamin D[25(OH)D],immune cell subsets,and helper T cell(Th)1 and Th2 in patients with unexplained recurrent spontaneous abortion(URSA).Methods A total of 307 URSA patients were enrolled as URSA group,which was further divided into non-maintenance of pregnancy group(n=127)and ma-intenance of pregnancy group(n=180)based on pregnancy maintenance status.Another 307 healthy pregnant women in the same period were selected as control group.The levels of 25(OH)D,immune cell subsets,and Th1/Th2 expression were compared among these groups.Pearson correlation analy-sis was used to explore the correlations of 25(OH)D and immune cell subsets with Th1/Th2 expres-sion in URSA patients.Results Compared with the control group,the URSA group had significantly lower levels of 25(OH)D and interleukin-10(IL-10),and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,interleukin-2(IL-2),and tumor necrosis fac-tor-α(TNF-α)(P＜0.001).Compared with the maintenance of pregnancy group,the non-maintenance of pregnancy group had significantly lower levels of 25(OH)D and IL-10,and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,IL-2,and TNF-α(P＜0.001).Pearson analysis revealed that 25(OH)D in URSA patients was significantly negatively correlated with IL-2 and TNF-α(P＜0.05),while positively correlated with IL-10(P＜0.05).Immune cell sub-sets,including CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,and CD19+were significantly positively correlated with IL-2 and TNF-α(P＜0.05),while negatively correlated with IL-10 in URSA patients(P＜0.05).Conclusion There are abnormalities in 25(OH)D,im-mune cell subsets,and Th1/Th2 cytokine expression in URSA patients,and these expressions differ in URSA patients with different pregnancy outcomes.The underlying mechanism may be related to the regulation of Th1/Th2 cytokine balance.

Strangles,caused by Streptococcus equi subsp.equi,remains one of the most prevalent and high-incidence infectious diseases in intensive donkey farms,posing a significant threat to the healthy development of the donkey industry.Vaccination serves as an effective measure for the pre-vention and control of the disease,however,there is currently no attenuated vaccine against this disease in China.To provide a candidate strain for the development of a live attenuated strangles vaccine,this study focused on a wild-type S.equi subsp.equi strain isolated from donkeys.Using homologous recombination gene knockout technology,the aroA gene(encoding 5-enolpyru-vylshikimate-3-phosphate synthase)and the sag A gene(encoding the precursor of streptolysin S toxin)were sequentially deleted to construct a double-gene-deletion strain(ΔsagA/aroA).The virulence and key biological characteristics of the mutant strain were systematically evaluated.TheΔsagA/aroA strain was successfully generated,exhibiting complete loss of hemolytic activity and maintaining stable genetic inheritance over 60 consecutive passages.Electron microscopy revealed that the mutant retained morphological characteristics compared to the wild-type strain,and its growth rate was significantly slower(P＜0.000 1).Virulence assessment using a challenge dose of 1× 105 CFU/0.2 mL(the minimum fully lethal dose of the wild-type strain)demonstrated markedly attenuated virulence in the mutant.Immunization trials with 1 ×104 CFU/0.2 mL of theΔsagA/aroA strain revealed a increase in ELISA antibody titers by day 7 post-vaccination,and higher levels at days 14 and 21.Notably,antibody levels in the experimental group were significant-ly higher than those in the control group(P＜0.000 1).These findings confirm that the double-gene-deletion strain S.equi subsp.equi ΔsagA/aroA exhibits reduced virulence while retaining im-munogenicity,which suggested it can be used as a promising candidate strain for further develop-ment of a live attenuated strangles vaccine.

Strangles,caused by Streptococcus equi subsp.equi,remains one of the most prevalent and high-incidence infectious diseases in intensive donkey farms,posing a significant threat to the healthy development of the donkey industry.Vaccination serves as an effective measure for the pre-vention and control of the disease,however,there is currently no attenuated vaccine against this disease in China.To provide a candidate strain for the development of a live attenuated strangles vaccine,this study focused on a wild-type S.equi subsp.equi strain isolated from donkeys.Using homologous recombination gene knockout technology,the aroA gene(encoding 5-enolpyru-vylshikimate-3-phosphate synthase)and the sag A gene(encoding the precursor of streptolysin S toxin)were sequentially deleted to construct a double-gene-deletion strain(ΔsagA/aroA).The virulence and key biological characteristics of the mutant strain were systematically evaluated.TheΔsagA/aroA strain was successfully generated,exhibiting complete loss of hemolytic activity and maintaining stable genetic inheritance over 60 consecutive passages.Electron microscopy revealed that the mutant retained morphological characteristics compared to the wild-type strain,and its growth rate was significantly slower(P＜0.000 1).Virulence assessment using a challenge dose of 1× 105 CFU/0.2 mL(the minimum fully lethal dose of the wild-type strain)demonstrated markedly attenuated virulence in the mutant.Immunization trials with 1 ×104 CFU/0.2 mL of theΔsagA/aroA strain revealed a increase in ELISA antibody titers by day 7 post-vaccination,and higher levels at days 14 and 21.Notably,antibody levels in the experimental group were significant-ly higher than those in the control group(P＜0.000 1).These findings confirm that the double-gene-deletion strain S.equi subsp.equi ΔsagA/aroA exhibits reduced virulence while retaining im-munogenicity,which suggested it can be used as a promising candidate strain for further develop-ment of a live attenuated strangles vaccine.

Objective:To investigate the efficacy and safety of bendamustine combined with anti-CD20 monoclonal antibody in the first-line treatment of older patients with indolent B-cell non-Hodgkin lymphoma (B-iNHL) .Methods:The clinical data of 159 patients with B-iNHL enrolled in 16 hospitals from Jiangsu Cooperative Lymphoma Group from December 1, 2019, to April 20, 2024, were analyzed for regimen efficacy and safety. Bendamustine plus rituximab (BR) and bendamustine plus obinutuzumab (BG) were administered to 139 (87.4% ) and 20 (12.6% ) patients, respectively.Results:Among the 159 patients, 101 (63.5% ) were male and 58 (36.5% ) were female, with a median age of 69 years (range: 60–84). Efficacy could be assessed in 138 (86.8% ) patients. The efficacy assessment demonstrated that the overall response rate was 92.0% with complete and partial remissions in 75 (54.3% ) and 52 (37.7% ) cases, respectively. With a median follow-up of 24 months (range: 4–64), the progression-free survival rate was (87.5 ± 3.0) % and the overall survival rate was (83.2 ± 3.3) %. Of the 27 patients who died, 6 (22.2% ) died due to disease progression. The mean applied dose of bendamustine per cycle was 73.0 (50.8–89.7) mg/m 2 per day, administered on days 1 and 2. Adverse events of grade 3 or higher were reported in 53 (33.3% ) patients, with infection (30 cases,18.9% ) and neutropenia (24 cases, 15.1% ) demonstrating the highest incidence. Conclusion:Bendamustine combined with anti-CD20 monoclonal antibody demonstrated good efficacy and is well-tolerated in the first-line treatment of elderly patients with B-iNHL.

Pancreatic cancer is among the most malignant cancers,and thus early intervention is the key to better survival outcomes.However,no methods have been derived that can reliably identify early precursors of development into malignancy.Therefore,it is urgent to discover early molecular changes during pancreatic tumorigenesis.As aberrant glycosylation is closely associated with cancer progression,numerous efforts have been made to mine glycosylation changes as biomarkers for diagnosis;however,detailed glycoproteomic information,especially site-specific N-glycosylation changes in pancreatic cancer with and without drug treatment,needs to be further explored.Herein,we used comprehensive solid-phase chemoenzymatic glycoproteomics to analyze glycans,glycosites,and intact glycopeptides in pancreatic cancer cells and patient sera.The profiling of N-glycans in cancer cells revealed an increase in the secreted glycoproteins from the primary tumor of MIA PaCa-2 cells,whereas human sera,which contain many secreted glycoproteins,had significant changes of glycans at their specific glycosites.These results indicated the potential role for tumor-specific glycosylation as disease biomarkers.We also found that AMG-510,a small molecule inhibitor against Kirsten rat sarcoma viral oncogene homolog(KRAS)G12C mutation,profoundly reduced the glycosylation level in MIA PaCa-2 cells,suggesting that KRAS plays a role in the cellular glycosylation process,and thus glycosylation inhibition contributes to the anti-tumor effect of AMG-510.

Objective The aim of this study was to use bioinformatics methods to integrate gene expression profiles and spli-cing profiles,and identify potential biomarkers that play a role in endocrine therapy resistance.Methods The gene datasets of endo-crine therapy sensitive and resistant cells from the Gene expression omnibus(GEO)database were downloaded to screen for differenti-ally expressed genes and differential splicing genes related to endocrine therapy resistance,and perform Hallmark enrichment analysis and Gene set enrichment analysis(GSEA)on these genes.The cancer genome atlas(TCGA)and the breast cancer data in the Molecu-lar taxonomy of breast cancer international consortium(METABRIC)were downloaded,and compared the expression of differential genes in tumor and adjacent tissues,as well as the differences between different tumor stages and grades.The relationship between dif-ferential expressed genes and overall survival(OS),alternative splicing,and immune infiltration were explored in patients receiving endocrine therapy.Results GEO data analysis showed that differentially expressed genes were mainly enriched in breast cancer treat-ment response related to pathways and immune-related pathways,and differentially spliced genes were mainly enriched in breast canc-er treatment response related to pathways.The genes that undergo differential expression and splicing simultaneously included CAMK2B,EVL,MZB1and PTPRG.Among them,the expression of MZB1 was associated with OS and immune cell infiltration in pa-tients receiving endocrine therapy,and the precursor mRNA of MZB1 underwent more alternative 3'splicing in drug-resistant cells.Conclusion The expression of MZB1 is related to the effect on endocrine therapy in breast cancer,and its mechanism may be the change of immune related function caused by alternative splicing.MZB1 can serve as a potential biomarker for endocrine therapy to improve prognosis.

Objective The aim of this study was to investigate the regulatory role and mechanism of TRIM24 in endocrine therapy resistance of breast cancer.Methods The relationship between TRIM24 expression and prognosis in breast cancer patients receiving endocrine therapy was analyzed by the Kaplan-Meier Plotter database.After treatment of breast cancer MCF7 cells or T47D cells with tamoxifen(TAM),the expression of TRIM24 was detected by qRT-PCR and Western blot.After knocking out TRIM24 in TAM sensitive MCF7/S0.5 cells and resistant cells MCF7/TAMR-1 cells,cell proliferation was measured by CCK 8 assay.After knocking out the estrogen receptor(ER)or treating with the selective progesterone receptor(PR)antagonist UPA(Ulipristal acetate)in MCF7 cells,the expression of TRIM24 was measured by qRT-PCR and Western blot.After PR was overexpressed in MCF7 cells,the expression of TRIM24 was detected by Western blot.The expression of human epidermal growth factor receptor 2(HER2)was detected by qRT PCR and Western blot after the overexpression of TRIM24.Results Among breast cancer patients receiving endocrine thera-py,patients with high expression of TRIM24 had a shorter recurrence free survival(RFS)(P=0.027).TAM treatment could induce an increase of TRIM24 expression(P<0.001);Knocking out TRIM24 significantly inhibited the proliferation of drug-resistant MCF7/TAMR-1 cells(P<0.001),but had no effect on sensitive MCF7/S0.5 cells;After knocking out ER or treatment of PR antagonist,the expression of TRIM24 could be upregulated in MCF7 cells(P<0.01),while overexpression of PR could down-regulate the expression of TRIM24.TRIM24 could activate the expression of HER2(P<0.05).Conclusion PR negatively regulation of TRIM24 plays an important role in endocrine therapy resistance of breast cancer.

Objective The aim of this study was to use bioinformatics methods to integrate gene expression profiles and spli-cing profiles,and identify potential biomarkers that play a role in endocrine therapy resistance.Methods The gene datasets of endo-crine therapy sensitive and resistant cells from the Gene expression omnibus(GEO)database were downloaded to screen for differenti-ally expressed genes and differential splicing genes related to endocrine therapy resistance,and perform Hallmark enrichment analysis and Gene set enrichment analysis(GSEA)on these genes.The cancer genome atlas(TCGA)and the breast cancer data in the Molecu-lar taxonomy of breast cancer international consortium(METABRIC)were downloaded,and compared the expression of differential genes in tumor and adjacent tissues,as well as the differences between different tumor stages and grades.The relationship between dif-ferential expressed genes and overall survival(OS),alternative splicing,and immune infiltration were explored in patients receiving endocrine therapy.Results GEO data analysis showed that differentially expressed genes were mainly enriched in breast cancer treat-ment response related to pathways and immune-related pathways,and differentially spliced genes were mainly enriched in breast canc-er treatment response related to pathways.The genes that undergo differential expression and splicing simultaneously included CAMK2B,EVL,MZB1and PTPRG.Among them,the expression of MZB1 was associated with OS and immune cell infiltration in pa-tients receiving endocrine therapy,and the precursor mRNA of MZB1 underwent more alternative 3'splicing in drug-resistant cells.Conclusion The expression of MZB1 is related to the effect on endocrine therapy in breast cancer,and its mechanism may be the change of immune related function caused by alternative splicing.MZB1 can serve as a potential biomarker for endocrine therapy to improve prognosis.

Objective The aim of this study was to investigate the regulatory role and mechanism of TRIM24 in endocrine therapy resistance of breast cancer.Methods The relationship between TRIM24 expression and prognosis in breast cancer patients receiving endocrine therapy was analyzed by the Kaplan-Meier Plotter database.After treatment of breast cancer MCF7 cells or T47D cells with tamoxifen(TAM),the expression of TRIM24 was detected by qRT-PCR and Western blot.After knocking out TRIM24 in TAM sensitive MCF7/S0.5 cells and resistant cells MCF7/TAMR-1 cells,cell proliferation was measured by CCK 8 assay.After knocking out the estrogen receptor(ER)or treating with the selective progesterone receptor(PR)antagonist UPA(Ulipristal acetate)in MCF7 cells,the expression of TRIM24 was measured by qRT-PCR and Western blot.After PR was overexpressed in MCF7 cells,the expression of TRIM24 was detected by Western blot.The expression of human epidermal growth factor receptor 2(HER2)was detected by qRT PCR and Western blot after the overexpression of TRIM24.Results Among breast cancer patients receiving endocrine thera-py,patients with high expression of TRIM24 had a shorter recurrence free survival(RFS)(P=0.027).TAM treatment could induce an increase of TRIM24 expression(P<0.001);Knocking out TRIM24 significantly inhibited the proliferation of drug-resistant MCF7/TAMR-1 cells(P<0.001),but had no effect on sensitive MCF7/S0.5 cells;After knocking out ER or treatment of PR antagonist,the expression of TRIM24 could be upregulated in MCF7 cells(P<0.01),while overexpression of PR could down-regulate the expression of TRIM24.TRIM24 could activate the expression of HER2(P<0.05).Conclusion PR negatively regulation of TRIM24 plays an important role in endocrine therapy resistance of breast cancer.