BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the un...BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the unclear mechanisms of proliferation and self-renewal in humans.AIM To investigate the role and mechanism of SPOC domain-containing protein 1(SPOCD1)in human SSC proliferation.METHODS We analyzed publicly available human testis single-cell RNA sequencing(RNAseq)data and found that SPOCD1 is predominantly expressed in SSCs in the early developmental stages.Small interfering RNA was applied to suppress SPOCD1 expression to detect the impacts of SPOCD1 inhibition on SSC proliferation and apoptosis.Subsequently,we explored the target genes of SPOCD1 using RNA-seq and confirmed their role by restoring the expression of the target genes.In addition,we examined SPOCD1 expression in some non-obstructive azoospermia(NOA)patients to explore the correlation between SPOCD1 and NOA.RESULTS The uniform manifold approximation and projection clustering and pseudotime analysis showed that SPOCD1 was highly expressed in the early stages of SSC,and immunohistological results showed that SPOCD1 was mainly localized in glial cell line-derived neurotrophic factor family receptor alpha-1 positive SSCs.SPOCD1 knockdown significantly inhibited cell proliferation and promoted apoptosis.RNA-seq results showed that SPOCD1 knockdown significantly downregulated genes such as adenylate kinase 4(AK4).Overexpression of AK4 in SPOCD1 knockdown cells partially reversed the phenotypic changes,indicating that AK4 is a functional target gene of SPOCD1.In addition,we found a significant downregulation of SPOCD1 expression in some NOA patients,suggesting that the downregulation of SPOCD1 may be relevant for NOA.CONCLUSION Our study broadens the understanding of human SSC fate determination and may offer new theories on the etiology of male infertility.展开更多
BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity...BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity response and hold extreme potential as efficient therapies for certain malignancies.However,ICIs with a single target exhibit poor overall response rate in hepatocellular carcinoma(HCC)patients due to the complex pathological mechanisms of HCC.AIM To investigate the effects of combined TIM-3 and PD-1 blockade on tumor development in an HCC mouse model,aiming to identify more effective immunotherapies and provide more treatment options for HCC patients.METHODS The levels of PD-1 and TIM-3 on CD4+and CD8+T cells from tumor tissues,ascites,and matched adjacent tissues from HCC patients were determined with flow cytometry.An HCC xenograft mouse model was established and treated with anti-TIM-3 monoclonal antibody(mAb)and/or anti-PD-1 mAb.Tumor growth in each group was measured.Hematoxylin and eosin staining and immunohistochemical staining were used to evaluate T cell infiltration in tumors.The percentage of CD4+and CD8+T cells in tissue samples from mice was tested with flow cytometry.The percentages of PD-1+CD8+,TIM-3+CD8+,and PD-1+TIM-3+CD8+T cells was accessed by flow cytometry.The levels of the cytokines including tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),interleukin(IL)-6,and IL-10 in tumor tissues were gauged with enzyme-linked immunosorbent assay kits.RESULTS We confirmed that PD-1 and TIM-3 expression was substantially upregulated in CD4+and CD8+T cells isolated from tumor tissues and ascites of HCC patients.TIM-3 mAb and PD-1 mAb treatment both reduced tumor volume and weight,while combined blockade had more substantial anti-tumor effects than individual treatment.Then we showed that combined therapy increased T cell infiltration into tumor tissues,and downregulated PD-1 and TIM-3 expression on CD8+T cells in tumor tissues.Moreover,combined treatment facilitated the production of T cell effector cytokines TNF-α and IFN-γ,and reduced the production of immunosuppressive cytokines IL-10 and IL-6 in tumor tissues.Thus,we implicated that combined blockade could ameliorate T cell exhaustion in HCC mouse model.CONCLUSION Combined TIM-3 and PD-1 blockade restrains HCC development by facilitating CD4+ and CD8+T cell-mediated antitumor immune responses.展开更多
AIM: To investigate if the presence of relevant genetic polymorphisms has effect on the effectual clearance of bacteria by monocytes and granulocytes in patients with Crohn’s disease (CD).
OBJECTIVE:To investigate the relationship between antiviral restriction factor Sterile Alpha Motif and Histidine-Aspartic acid domain-containing protein 1(SAMHD1)expression and T cell activation,furthermore,identifyin...OBJECTIVE:To investigate the relationship between antiviral restriction factor Sterile Alpha Motif and Histidine-Aspartic acid domain-containing protein 1(SAMHD1)expression and T cell activation,furthermore,identifying objective indexes of lung-spleen Qi deficiency symptom pattern.METHODS:We assessed the profile of T lymphocyte subsets,characteristics of SAMHD1 and human leukocyte antigen DR(HLA-DR)expression in lungspleen Qi deficiency patients.At the same time,people living with human immunodeficiency virus/acquired immune deficiency syndrome(HIV/AIDS)(PLWHA)without obvious clinical symptoms and healthy donors in this area were used as controls.RESULTS:Immunohematologic indexes lower CD4 count,lower CD4/CD8 ratio and higher SAMHD1 level were found in lung-spleen Qi deficiency patients.Furthermore,we demonstrated a positive relationship between SAMHD1 and HLA-DR level as well as with interferon factor in lung-spleen Qi deficiency syndrome and patients without obvious clinical signs and symptoms groups.CONCLUSIONS:These data indicated the positive relationship between SAMHD1 and T cell activation which further elucidated the role of SAMHD1 in cellular immune response.Furthermore,combination of T lymphocyte subsets counts and SAMHD1 level may be used as clinical and biological reference basis for the differentiation and diagnosis of HIV/AIDS traditional Chinese medicine syndromes.展开更多
目的探讨急性缺血性脑卒中患者静脉溶栓前后血浆信号肽Cub-Egf结构域蛋白1(Plasma signal peptide Cub Egf domain protein 1,SCUBE1)水平变化及血浆SCUBE1水平对患者预后不良的预测价值。方法纳入2020年1月-2022年1月本院收治的116例...目的探讨急性缺血性脑卒中患者静脉溶栓前后血浆信号肽Cub-Egf结构域蛋白1(Plasma signal peptide Cub Egf domain protein 1,SCUBE1)水平变化及血浆SCUBE1水平对患者预后不良的预测价值。方法纳入2020年1月-2022年1月本院收治的116例急性缺血性脑卒中患者作为研究对象,另选取同期体检健康者120例作为对照组,收集患者临床资料,测定研究对象血浆SCUBE1水平,比较患者与对照组以及患者治疗不同时间点(治疗前、治疗2、12、24 h、3 d后)血浆SCUBE1水平,比较不同预后情况患者临床资料以及血浆SCUBE1水平差异,用受试者工作特征(Receiver operating characteristic,ROC)曲线分析血浆SCUBE1水平对急性缺血性脑卒中患者静脉溶栓后预后不良的预测价值。结果与对照组比较,急性脑卒中组患者血浆SCUBE1水平升高(P<0.05);与治疗前比较,治疗2、12、24 h后血浆SCUBE1水平升高(P<0.05),治疗3 d后血浆SCUBE1水平降低(P<0.05);与治疗12 h后比较,治疗24 h、3 d后血浆SCUBE1水平降低(P<0.05);预后不良组患者高血压病、高脂血症、糖尿病、心房颤动比例、基线美国国立卫生院卒中量表(National institutes of health stroke scale,NIHSS)评分、血浆SCUBE1水平高于预后良好组(P<0.05);基线NIHSS评分、血浆SCUBE1水平是患者预后不良的独立危险因素(P<0.05);血浆SCUBE1水平预测急性脑卒中患者静脉溶栓后预后不良的曲线下面积(Area under curve,AUC)为0.860(敏感度为73.0%,特异性为88.6%)。结论急性缺血性脑卒中患者血浆SCUBE1水平升高,经静脉溶栓后水平降低,患者治疗前血浆SCUBE1水平有望成为患者短期预后不良的预测因子。展开更多
目的探讨CUB结构域蛋白1（CUB domain-containing protein 1,CDCP1）在肺癌中的表达及临床意义。方法肺癌患者78例,其中40例采用Western blot方法检测肺癌及手术切缘癌旁组织中CDCP1蛋白表达;应用免疫组织化学法检测78例肺癌及切缘组织...目的探讨CUB结构域蛋白1（CUB domain-containing protein 1,CDCP1）在肺癌中的表达及临床意义。方法肺癌患者78例,其中40例采用Western blot方法检测肺癌及手术切缘癌旁组织中CDCP1蛋白表达;应用免疫组织化学法检测78例肺癌及切缘组织病理切片中CDCP1的表达,并分析其与肺癌生物学行为的关系。结果肺癌组织中CDCP1高表达率（67.5%）高于癌旁组织（32.5%）,随TNM分期增加,CDCP1在肺癌组织中表达量逐渐增加（P〈0.05）;免疫组织化学结果显示,CDCP1在肺癌组织中高表达率（57.7%）高于癌旁组织（14.1%）（P〈0.05）;有淋巴结转移者CDCP1表达率（67.3%）高于无淋巴结转移者（38.5%）（P〈0.05）;CDCP1表达与肺癌患者生存期呈负相关（r=0.804,P=0.031）;Cox分析显示TNM分期（OR=1.216,95%CI：0.649～5.525,P=0.012）和CDCP1（OR=0.526,95%CI：0.153～0.426,P=0.009）是影响肺癌患者预后的独立因素。结论 CDCP1可能是肺癌多阶段演变过程中的重要变化分子之一,可能成为肺癌分子诊断的生物标志物。展开更多
Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogen...Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.展开更多
基金the National Natural Science Foundation for Young Scholars of China,No.82201771National Natural Science Foundation of China,No.32270912+2 种基金Natural Science Foundation of Changsha,No.kq2202491Research Grant of CITIC-Xiangya,No.YNXM202109 and No.YNXM202115Hunan Provincial Grant for Innovative Province Construction,No.2019SK4012。
文摘BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the unclear mechanisms of proliferation and self-renewal in humans.AIM To investigate the role and mechanism of SPOC domain-containing protein 1(SPOCD1)in human SSC proliferation.METHODS We analyzed publicly available human testis single-cell RNA sequencing(RNAseq)data and found that SPOCD1 is predominantly expressed in SSCs in the early developmental stages.Small interfering RNA was applied to suppress SPOCD1 expression to detect the impacts of SPOCD1 inhibition on SSC proliferation and apoptosis.Subsequently,we explored the target genes of SPOCD1 using RNA-seq and confirmed their role by restoring the expression of the target genes.In addition,we examined SPOCD1 expression in some non-obstructive azoospermia(NOA)patients to explore the correlation between SPOCD1 and NOA.RESULTS The uniform manifold approximation and projection clustering and pseudotime analysis showed that SPOCD1 was highly expressed in the early stages of SSC,and immunohistological results showed that SPOCD1 was mainly localized in glial cell line-derived neurotrophic factor family receptor alpha-1 positive SSCs.SPOCD1 knockdown significantly inhibited cell proliferation and promoted apoptosis.RNA-seq results showed that SPOCD1 knockdown significantly downregulated genes such as adenylate kinase 4(AK4).Overexpression of AK4 in SPOCD1 knockdown cells partially reversed the phenotypic changes,indicating that AK4 is a functional target gene of SPOCD1.In addition,we found a significant downregulation of SPOCD1 expression in some NOA patients,suggesting that the downregulation of SPOCD1 may be relevant for NOA.CONCLUSION Our study broadens the understanding of human SSC fate determination and may offer new theories on the etiology of male infertility.
基金Supported by the First-Class Discipline Construction Founded Project of Ningxia Medical University and the School of Clinical Medicine,No.2020008.
文摘BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity response and hold extreme potential as efficient therapies for certain malignancies.However,ICIs with a single target exhibit poor overall response rate in hepatocellular carcinoma(HCC)patients due to the complex pathological mechanisms of HCC.AIM To investigate the effects of combined TIM-3 and PD-1 blockade on tumor development in an HCC mouse model,aiming to identify more effective immunotherapies and provide more treatment options for HCC patients.METHODS The levels of PD-1 and TIM-3 on CD4+and CD8+T cells from tumor tissues,ascites,and matched adjacent tissues from HCC patients were determined with flow cytometry.An HCC xenograft mouse model was established and treated with anti-TIM-3 monoclonal antibody(mAb)and/or anti-PD-1 mAb.Tumor growth in each group was measured.Hematoxylin and eosin staining and immunohistochemical staining were used to evaluate T cell infiltration in tumors.The percentage of CD4+and CD8+T cells in tissue samples from mice was tested with flow cytometry.The percentages of PD-1+CD8+,TIM-3+CD8+,and PD-1+TIM-3+CD8+T cells was accessed by flow cytometry.The levels of the cytokines including tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),interleukin(IL)-6,and IL-10 in tumor tissues were gauged with enzyme-linked immunosorbent assay kits.RESULTS We confirmed that PD-1 and TIM-3 expression was substantially upregulated in CD4+and CD8+T cells isolated from tumor tissues and ascites of HCC patients.TIM-3 mAb and PD-1 mAb treatment both reduced tumor volume and weight,while combined blockade had more substantial anti-tumor effects than individual treatment.Then we showed that combined therapy increased T cell infiltration into tumor tissues,and downregulated PD-1 and TIM-3 expression on CD8+T cells in tumor tissues.Moreover,combined treatment facilitated the production of T cell effector cytokines TNF-α and IFN-γ,and reduced the production of immunosuppressive cytokines IL-10 and IL-6 in tumor tissues.Thus,we implicated that combined blockade could ameliorate T cell exhaustion in HCC mouse model.CONCLUSION Combined TIM-3 and PD-1 blockade restrains HCC development by facilitating CD4+ and CD8+T cell-mediated antitumor immune responses.
文摘AIM: To investigate if the presence of relevant genetic polymorphisms has effect on the effectual clearance of bacteria by monocytes and granulocytes in patients with Crohn’s disease (CD).
基金Supported by the National Natural Science Foundation of China(Construction of HIV Pseudovirus Infected Cell Model and Application of Yiaikang in the Study of T Cell Immune Response,No.82004207)the National Special Science and Technology Program on Major Infectious Diseases(Study on the Scheme of TCM to Reduce the Incidence of Adverse Reactions to AIDS Antiviral Therapy,No.2017ZX10205502002+6 种基金Study on the Scheme of TCM to Reduce the Incidence of AIDS Opportunistic Infection,No.2017ZX10205502003)the Science and Technology Tackling Plan of Henan Province(Exploring the Activation Mechanism of Yiaikang on CD8+T Lymphocytes in HIV/AIDS Patients based on STING/SAMHD1 Pathway,No.202102310178Regulation of HIV/AIDS Treg CellDerived Exosomeso on Activation of CD4+T Cells and the Intervention of Yiaikang,No.212102311126)Subject Construction Project of Henan Characteristic Backbone Subject of TCM of Henan University of TCM(Deep Sequencing/Molecular Docking/Gene Editing Network System to Analyze The Mechanism of TCM Regulating the Immune Function of HIV Patients,No.STG-ZYXKY-2020029)Henan Province TCM Research Project(Explore the Mechanism of Yiaikang in the Treatment of AIDS Immunity Based on HIV gag-CTL Response,No.2018JDZX106,2019AZB003Study on TCM Treatment of Aged AIDS,No.20-21ZYZD03)Key Scientific Research Projects of Colleges and Universities in Henan Province(to Explore the Regulation Mechanism of Yiaikang on AIDS Lung Qi Deficiency Syndrome from TGF-β/Smad/ID,No.20A360010)
文摘OBJECTIVE:To investigate the relationship between antiviral restriction factor Sterile Alpha Motif and Histidine-Aspartic acid domain-containing protein 1(SAMHD1)expression and T cell activation,furthermore,identifying objective indexes of lung-spleen Qi deficiency symptom pattern.METHODS:We assessed the profile of T lymphocyte subsets,characteristics of SAMHD1 and human leukocyte antigen DR(HLA-DR)expression in lungspleen Qi deficiency patients.At the same time,people living with human immunodeficiency virus/acquired immune deficiency syndrome(HIV/AIDS)(PLWHA)without obvious clinical symptoms and healthy donors in this area were used as controls.RESULTS:Immunohematologic indexes lower CD4 count,lower CD4/CD8 ratio and higher SAMHD1 level were found in lung-spleen Qi deficiency patients.Furthermore,we demonstrated a positive relationship between SAMHD1 and HLA-DR level as well as with interferon factor in lung-spleen Qi deficiency syndrome and patients without obvious clinical signs and symptoms groups.CONCLUSIONS:These data indicated the positive relationship between SAMHD1 and T cell activation which further elucidated the role of SAMHD1 in cellular immune response.Furthermore,combination of T lymphocyte subsets counts and SAMHD1 level may be used as clinical and biological reference basis for the differentiation and diagnosis of HIV/AIDS traditional Chinese medicine syndromes.
文摘目的探讨急性缺血性脑卒中患者静脉溶栓前后血浆信号肽Cub-Egf结构域蛋白1(Plasma signal peptide Cub Egf domain protein 1,SCUBE1)水平变化及血浆SCUBE1水平对患者预后不良的预测价值。方法纳入2020年1月-2022年1月本院收治的116例急性缺血性脑卒中患者作为研究对象,另选取同期体检健康者120例作为对照组,收集患者临床资料,测定研究对象血浆SCUBE1水平,比较患者与对照组以及患者治疗不同时间点(治疗前、治疗2、12、24 h、3 d后)血浆SCUBE1水平,比较不同预后情况患者临床资料以及血浆SCUBE1水平差异,用受试者工作特征(Receiver operating characteristic,ROC)曲线分析血浆SCUBE1水平对急性缺血性脑卒中患者静脉溶栓后预后不良的预测价值。结果与对照组比较,急性脑卒中组患者血浆SCUBE1水平升高(P<0.05);与治疗前比较,治疗2、12、24 h后血浆SCUBE1水平升高(P<0.05),治疗3 d后血浆SCUBE1水平降低(P<0.05);与治疗12 h后比较,治疗24 h、3 d后血浆SCUBE1水平降低(P<0.05);预后不良组患者高血压病、高脂血症、糖尿病、心房颤动比例、基线美国国立卫生院卒中量表(National institutes of health stroke scale,NIHSS)评分、血浆SCUBE1水平高于预后良好组(P<0.05);基线NIHSS评分、血浆SCUBE1水平是患者预后不良的独立危险因素(P<0.05);血浆SCUBE1水平预测急性脑卒中患者静脉溶栓后预后不良的曲线下面积(Area under curve,AUC)为0.860(敏感度为73.0%,特异性为88.6%)。结论急性缺血性脑卒中患者血浆SCUBE1水平升高,经静脉溶栓后水平降低,患者治疗前血浆SCUBE1水平有望成为患者短期预后不良的预测因子。
文摘Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.
