Background:This study was designed to investigate the feasibility of tumor-infiltrating immune cells with different phenotypic characteristics for predicting short-term clinical responses in patients with locally adva...Background:This study was designed to investigate the feasibility of tumor-infiltrating immune cells with different phenotypic characteristics for predicting short-term clinical responses in patients with locally advanced cervical cancer(LACC).Methods:Thirty-four patients who received concurrent chemoradiotherapy and twenty-one patients who merely underwent radiotherapy were enrolled in this study.We retrospectively analyzed the T cell markers(i.e.,CD3,CD4,CD8),memory markers(i.e.,CD45,CCR7),and differentiation markers(i.e.,CD27)in the peripheral blood and tumor tissues of patients with LACC before treatment based on flow cytometry.We also analyzed the relationship of T cell subsets between peripheral blood and tumor tissues,and their correlation with complete response or partial response.Results:The percentage of central memory CD8^(+)TCM(CD8^(+)CD45RA^(−)CD27^(+)CCR7^(+))cells in LACC patients was significantly lower than that of the control group.The percentage of CD8^(+)TN in the peripheral blood of LACC patients was significantly higher than that of tumor tissues.CD8^(+)TEM in the peripheral blood was significantly lower than that of tumor tissues.The percentage of CD8^(+)TN and CD8^(+)TCM in human papillomavirus(HPV)positive samples was significantly higher than that of HPV-negative samples.Similarly,the percentage of CD8^(+)TCM in tumor tissues was significantly higher in cancer tissue samples with lymph nodes compared with those without.Conclusion:A higher proportion of CD4^(+)TCM and a lower proportion of CD8^(+)TN in the tumor microenvironment of LACC may contribute to the therapy response prediction.展开更多
Background:Cervical squamous cell carcinoma(CESC),the most common subtype of cervical cancer,is primarily caused by the high-risk human papillomavirus(HPV)infection and genetic susceptibility.Single-cell RNA sequencin...Background:Cervical squamous cell carcinoma(CESC),the most common subtype of cervical cancer,is primarily caused by the high-risk human papillomavirus(HPV)infection and genetic susceptibility.Single-cell RNA sequencing(scRNA-seq)has been widely used in CESC research to uncover the diversity of cell types and states within tumor tissues,enabling a detailed study of the tumor microenvironment(TME).This technology allows precise mapping of HPV infection in cervical tissues,providing valuable insights into the initiation and progression of HPV-mediated malignant transformation.Methods:We performed the scRNA-seq to characterize gene expression in tumor tissues and paired adjacent para-cancerous tissues from four patients with early-stage CESC using the 10×Genomics platform.The HPV infection and its subtypes were identified using the scRNA data and viral sequence mapping,and trajectory analyses were performed using HPV+or HPV-cells.Interactions between different types of keratinized cells and their interactions with other cell types were identified,and pathways and specificity markers were screened for proliferating keratinized cells.The Cancer Genome Atlas(TCGA)dataset was used to verify the prognostic correlation between tumor-specific PI3+S100A7+keratinocyte infiltration and CESC,and the localization relationship between PI3+S100A7+keratinocytes and macrophages was verified by immunofluorescence staining.Results:Various types of keratinocytes and fibroblasts were the two cell types with the most significant differences in percentage between the tumor tissue samples and paired adjacent non-cancerous tissue samples in the early stages of CESC.We found that PI3+S100A7+keratinocytes were associated with early HPV-positive CESC,and PI3+S100A7+keratinocytes were more abundant in tumors than in adjacent normal tissues in the TCGA-CESC dataset.Analysis of clinical information revealed that the infiltration of PI3+S100A7+keratinocytes was notably higher in tumors with poor prognosis than in those with good prognosis.Additionally,multiplex immunofluorescence analysis showed a specific increase in PI3+S100A7+expression within tumor tissues,with PI3+S100A7+keratinocytes and CD163+macrophages being spatially very close to each other.In the analysis of cell-cell interactions,macrophages exhibited strong crosstalk with PI3+S100A7+proliferating keratinocytes in HPV-positive CESC tumors,mediated by tumor necrosis factor(TNF),CCL2,CXCL8,and IL10,highlighting the dynamic and tumor-specific enhancement of macrophage-keratinocyte interactions,which are associated with poor prognosis and immune modulation.Using CIBERSORTx,we discovered that patients with high infiltration of both PI3+S100A7+proliferating keratinocytes and macrophages had the shortest overall survival.In the analysis of cell-cell interactions,PI3+S100A7+proliferating keratinocytes and macrophages were found to be involved in highly active pathways that promote differentiation and structure formation,including cytokine receptor interactions,the Nuclear Factor kappa-light-chain-enhancer of activated B cells(NF-κB)signaling pathway,and TNF signaling pathway regulation.Further subtyping of fibroblast populations identified four subtypes.The C1 group,characterized by its predominance in tumor tissues,is a subtype enriched with cancer-associated fibroblasts(CAFs),whereas the C3 group is primarily enriched in adjacent non-cancerous tissues and consists of undifferentiated cells.Moreover,the distinct molecular and cellular differences between HPV16-and HPV66-associated tumors were demonstrated,emphasizing the unique tumor-promoting mechanisms and microenvironmental influences driven by each HPV subtype.Conclusions:We discovered a heterogeneous population of keratinocytes between tumor and adjacent non-cancerous tissues caused by HPV infection and identified macrophages and specific CAFs that play a crucial role during the early stage in promoting the inflammatory response and remodeling the cancer-promoting TME.Our findings provide new insights into the transcriptional landscape of early-stage CESC to understand the mechanism of HPV-mediated malignant transformation in cervical cancer.展开更多
For patients with lung disease,dry powder inhalers(DPI)are profoundly beneficial.The current study introduces and develops a series of dry powder inhalers(DPIs).A capsule-based(size 0)active DPI was considered.The stu...For patients with lung disease,dry powder inhalers(DPI)are profoundly beneficial.The current study introduces and develops a series of dry powder inhalers(DPIs).A capsule-based(size 0)active DPI was considered.The study aims to investigate whether swirling flow and outlet capillary diameter(dc_out)affect the percentage of emitted doses(ED)released from the capsule.Spiral vanes were added to the capillary inlet to produce a swirling flow.Computational fluid dynamics(CFD)was applied to simulate the problem.The results were compared with previous in vitro and numerical studies to validate the results.Based on the derived results,the small swirl parameter(SP)enhances the secondary flow and recirculation zone.It increases the central jet flow,which increases the ED value by about 5–20%compared to no-swirl flow.However,as the airflow rate increases,the recirculation zone enlarges,vorticities become dominant,and asymmetrical flow patterns emerge.Consequently,ED%drops significantly(more than 50%).As d_(c_out)decreases,the vorticities around the outlet capillary become more potent,which is undesirable.Indeed,the emptying of the capsule does not happen ideally.The research provides a perspective on the device's design and DPI performance.展开更多
基金the Project of the Central Government Guiding Local Science and Technology under Grant Number ZYYD2022B18the Institutional Ethics Committee of Affiliated Cancer Hospital of Xinjiang Medical University(No.K-2019001).
文摘Background:This study was designed to investigate the feasibility of tumor-infiltrating immune cells with different phenotypic characteristics for predicting short-term clinical responses in patients with locally advanced cervical cancer(LACC).Methods:Thirty-four patients who received concurrent chemoradiotherapy and twenty-one patients who merely underwent radiotherapy were enrolled in this study.We retrospectively analyzed the T cell markers(i.e.,CD3,CD4,CD8),memory markers(i.e.,CD45,CCR7),and differentiation markers(i.e.,CD27)in the peripheral blood and tumor tissues of patients with LACC before treatment based on flow cytometry.We also analyzed the relationship of T cell subsets between peripheral blood and tumor tissues,and their correlation with complete response or partial response.Results:The percentage of central memory CD8^(+)TCM(CD8^(+)CD45RA^(−)CD27^(+)CCR7^(+))cells in LACC patients was significantly lower than that of the control group.The percentage of CD8^(+)TN in the peripheral blood of LACC patients was significantly higher than that of tumor tissues.CD8^(+)TEM in the peripheral blood was significantly lower than that of tumor tissues.The percentage of CD8^(+)TN and CD8^(+)TCM in human papillomavirus(HPV)positive samples was significantly higher than that of HPV-negative samples.Similarly,the percentage of CD8^(+)TCM in tumor tissues was significantly higher in cancer tissue samples with lymph nodes compared with those without.Conclusion:A higher proportion of CD4^(+)TCM and a lower proportion of CD8^(+)TN in the tumor microenvironment of LACC may contribute to the therapy response prediction.
基金supported by grants from the Shanghai Cooperation Organization Science and Technology Partnership Program and the International Science and Technology Cooperation Program(No.2020E01056)the Tianshan Talent Program-Science and Technology Innovation Team(No.2023TSYCTD0015).
文摘Background:Cervical squamous cell carcinoma(CESC),the most common subtype of cervical cancer,is primarily caused by the high-risk human papillomavirus(HPV)infection and genetic susceptibility.Single-cell RNA sequencing(scRNA-seq)has been widely used in CESC research to uncover the diversity of cell types and states within tumor tissues,enabling a detailed study of the tumor microenvironment(TME).This technology allows precise mapping of HPV infection in cervical tissues,providing valuable insights into the initiation and progression of HPV-mediated malignant transformation.Methods:We performed the scRNA-seq to characterize gene expression in tumor tissues and paired adjacent para-cancerous tissues from four patients with early-stage CESC using the 10×Genomics platform.The HPV infection and its subtypes were identified using the scRNA data and viral sequence mapping,and trajectory analyses were performed using HPV+or HPV-cells.Interactions between different types of keratinized cells and their interactions with other cell types were identified,and pathways and specificity markers were screened for proliferating keratinized cells.The Cancer Genome Atlas(TCGA)dataset was used to verify the prognostic correlation between tumor-specific PI3+S100A7+keratinocyte infiltration and CESC,and the localization relationship between PI3+S100A7+keratinocytes and macrophages was verified by immunofluorescence staining.Results:Various types of keratinocytes and fibroblasts were the two cell types with the most significant differences in percentage between the tumor tissue samples and paired adjacent non-cancerous tissue samples in the early stages of CESC.We found that PI3+S100A7+keratinocytes were associated with early HPV-positive CESC,and PI3+S100A7+keratinocytes were more abundant in tumors than in adjacent normal tissues in the TCGA-CESC dataset.Analysis of clinical information revealed that the infiltration of PI3+S100A7+keratinocytes was notably higher in tumors with poor prognosis than in those with good prognosis.Additionally,multiplex immunofluorescence analysis showed a specific increase in PI3+S100A7+expression within tumor tissues,with PI3+S100A7+keratinocytes and CD163+macrophages being spatially very close to each other.In the analysis of cell-cell interactions,macrophages exhibited strong crosstalk with PI3+S100A7+proliferating keratinocytes in HPV-positive CESC tumors,mediated by tumor necrosis factor(TNF),CCL2,CXCL8,and IL10,highlighting the dynamic and tumor-specific enhancement of macrophage-keratinocyte interactions,which are associated with poor prognosis and immune modulation.Using CIBERSORTx,we discovered that patients with high infiltration of both PI3+S100A7+proliferating keratinocytes and macrophages had the shortest overall survival.In the analysis of cell-cell interactions,PI3+S100A7+proliferating keratinocytes and macrophages were found to be involved in highly active pathways that promote differentiation and structure formation,including cytokine receptor interactions,the Nuclear Factor kappa-light-chain-enhancer of activated B cells(NF-κB)signaling pathway,and TNF signaling pathway regulation.Further subtyping of fibroblast populations identified four subtypes.The C1 group,characterized by its predominance in tumor tissues,is a subtype enriched with cancer-associated fibroblasts(CAFs),whereas the C3 group is primarily enriched in adjacent non-cancerous tissues and consists of undifferentiated cells.Moreover,the distinct molecular and cellular differences between HPV16-and HPV66-associated tumors were demonstrated,emphasizing the unique tumor-promoting mechanisms and microenvironmental influences driven by each HPV subtype.Conclusions:We discovered a heterogeneous population of keratinocytes between tumor and adjacent non-cancerous tissues caused by HPV infection and identified macrophages and specific CAFs that play a crucial role during the early stage in promoting the inflammatory response and remodeling the cancer-promoting TME.Our findings provide new insights into the transcriptional landscape of early-stage CESC to understand the mechanism of HPV-mediated malignant transformation in cervical cancer.
基金This work was supported the National Natural Science Foundation of China(grant No.12172146).
文摘For patients with lung disease,dry powder inhalers(DPI)are profoundly beneficial.The current study introduces and develops a series of dry powder inhalers(DPIs).A capsule-based(size 0)active DPI was considered.The study aims to investigate whether swirling flow and outlet capillary diameter(dc_out)affect the percentage of emitted doses(ED)released from the capsule.Spiral vanes were added to the capillary inlet to produce a swirling flow.Computational fluid dynamics(CFD)was applied to simulate the problem.The results were compared with previous in vitro and numerical studies to validate the results.Based on the derived results,the small swirl parameter(SP)enhances the secondary flow and recirculation zone.It increases the central jet flow,which increases the ED value by about 5–20%compared to no-swirl flow.However,as the airflow rate increases,the recirculation zone enlarges,vorticities become dominant,and asymmetrical flow patterns emerge.Consequently,ED%drops significantly(more than 50%).As d_(c_out)decreases,the vorticities around the outlet capillary become more potent,which is undesirable.Indeed,the emptying of the capsule does not happen ideally.The research provides a perspective on the device's design and DPI performance.
