目的:探讨血清基质金属蛋白酶-1(matrix metalloproteinase-1,MMP1)和纽约食管鳞状细胞癌抗原-1(New York esophageal squamous cell carcinoma 1,NY-ESO-1)自身抗体联合检测在食管鳞状细胞癌中的诊断意义。方法:应用酶联免疫吸附实验检...目的:探讨血清基质金属蛋白酶-1(matrix metalloproteinase-1,MMP1)和纽约食管鳞状细胞癌抗原-1(New York esophageal squamous cell carcinoma 1,NY-ESO-1)自身抗体联合检测在食管鳞状细胞癌中的诊断意义。方法:应用酶联免疫吸附实验检测120例食管鳞状细胞癌患者和120例正常对照血清中MMP1和NY-ESO-1自身抗体的表达水平,采用受试者工作特征(receiver operating characteristic,ROC)曲线评价诊断效能。结果:血清MMP1和NY-ESO-1自身抗体在食管鳞状细胞癌患者中的表达均明显高于正常对照[(8.070±5.738)ng/mL vs(4.331±3.137)ng/mL,Z=6.214,P<0.001;0.463±0.571 vs 0.156±0.086,Z=5.210,P<0.001]。ROC曲线显示,当血清MMP1为最佳诊断临界值10.586 ng/mL时,其在诊断食管鳞状细胞癌的曲线下面积(area under the ROC curve,AUC)为0.732(95%CI:0.671~0.787),敏感度为24.2%,特异度为95.0%。NY-ESO-1自身抗体诊断食管鳞状细胞癌AUC为0.695(95%CI:0.632~0.752),敏感度为33.0%,特异度为95.0%。MMP1和NY-ESO-1自身抗体联合检测诊断食管鳞状细胞癌的AUC为0.800(95%CI:0.744~0.849),敏感度为47.5%,特异度为95.0%。结论:血清MMP1和NY-ESO-1自身抗体联合检测可能有助于提高食管鳞状细胞癌的诊断效能。展开更多
BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),t...BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),the mechanisms are largely unknown.AIM To explore the role of BMI-1,a polycomb group protein family member,which is highly expressed in LM,and the interaction between aHSCs and CRC cells in promoting CRC liver metastasis(CRLM).METHODS Immunohistochemistry was carried out to examine BMI-1 expression in LM and matched liver specimens of CRC.The expression levels of BMI-1 in mouse liver during CRLM(0,7,14,21,and 28 d)were detected by Western blotting(WB)and the quantitative polymerase chain reaction(qPCR)assay.We overexpressed BMI-1 in HSCs(LX2)by lentivirus infection and tested the molecular markers of aHSCs by WB,qPCR,and the immunofluorescence assay.CRC cells(HCT116 and DLD1)were cultured in HSC-conditioned medium(LX2 NC CM or LX2 BMI-1 CM).CM-induced CRC cell proliferation,migration,epithelial-mesenchymal transition(EMT)phenotype,and transforming growth factor beta(TGF-β)/SMAD pathway changes were investigated in vitro.A mouse subcutaneous xenotransplantation tumor model was established by co-implantation of HSCs(LX2 NC or LX2 BMI-1)and CRC cells to investigate the effects of HSCs on tumor growth and the EMT phenotype in vivo.RESULTS Positive of BMI-1 expression in the liver of CRLM patients was 77.8%.The expression level of BMI-1 continued to increase during CRLM in mouse liver cells.LX2 overexpressed BMI-1 was activated,accompanied by increased expression level of alpha smooth muscle actin,fibronectin,TGF-β1,matrix metalloproteinases,and interleukin 6.CRC cells cultured in BMI-1 CM exhibited enhanced proliferation and migration ability,EMT phenotype and activation of the TGF-β/SMAD pathway.In addition,the TGF-βR inhibitor SB-505124 diminished the effect of BMI-1 CM on SMAD2/3 phosphorylation in CRC cells.Furthermore,BMI-1 overexpressed LX2 HSCs promoted tumor growth and the EMT phenotype in vivo.CONCLUSION High expression of BMI-1 in liver cells is associated with CRLM progression.BMI-1 activates HSCs to secrete factors to form a prometastatic environment in the liver,and aHSCs promote proliferation,migration,and the EMT in CRC cells partially through the TGF-β/SMAD pathway.展开更多
目的:研究微小RNA-218(microRNA-218,miR-218)靶向B细胞特性莫洛尼鼠白血病病毒插入位点1(B-cell specific moloney leukemia virus insertion site 1,Bmi-1)抑制肺腺癌细胞增殖的作用及机制。方法:培养肺腺癌细胞株A549,分别转染miR-...目的:研究微小RNA-218(microRNA-218,miR-218)靶向B细胞特性莫洛尼鼠白血病病毒插入位点1(B-cell specific moloney leukemia virus insertion site 1,Bmi-1)抑制肺腺癌细胞增殖的作用及机制。方法:培养肺腺癌细胞株A549,分别转染miR-阴性对照(NC)核苷酸序列、miR-218核苷酸序列、NC质粒、Bmi-1质粒,采用平板克隆形成实验、CCK8实验检测细胞增殖水平,采用荧光定量PCR实验检测miR-218的表达,采用Western blot实验检测Bmi-1的表达,采用双荧光素酶报告基因实验检测miR-218与Bmi-1的靶向关系。建立移植瘤裸鼠,慢病毒转染miR-NC或miR-218核苷酸序列,检测移植瘤体积、质量及Bmi-1的表达。结果:与转染miR-NC核苷酸序列的miR-NC组比较,转染miR-218核苷酸序列的miR-218组A549细胞miR-218表达水平、增殖抑制率增加,克隆形成率、Bmi-1表达水平及野生型Bmi-1质粒的荧光素酶活性降低(P<0.05);与共转染miR-218核苷酸序列及NC质粒的miR-218+NC质粒组比较,共转染miR-218核苷酸序列及Bmi-1质粒的miR-218+Bmi-1质粒组A549细胞增殖抑制率降低,克隆形成率、Bmi-1表达水平增加(P<0.05);与慢病毒转染miR-NC核苷酸序列的miR-NC组比较,慢病毒转染miR-218核苷酸序列的miR-218组裸鼠移植瘤体积、质量及Bmi-1表达水平均降低(P<0.05)。结论:miR-218靶向下调Bmi-1表达并抑制肺腺癌细胞增殖。展开更多
BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by th...BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by the infiltration of neutrophils in the inflamed mucosa and the accumulation of pathogens.Like neutrophils in the context of innate immune responses,immunoglobulin A(IgA)as an acquired immune response partakes in the defense of the intestinal epithelium.Under normal conditions,IgA contributes to the elimination of microbes,but in connection with the loss of tolerance to chitinase 3-like 1(CHI3L1)in IBD,IgA could participate in CHI3L1-mediated improved adhesion and invasion of potentially pathogenic microorganisms.The tolerance brake to CHI3L1 and the occurrence of IgA autoantibodies to this particular target,the exact role and underlying mechanisms of CHI3L1 in the pathogenesis of IBD are still unclear.AIM To determine the predictive potential of Ig subtypes of a novel serological marker,anti-CHI3L1 autoantibodies(aCHI3L1)in determining the disease phenotype,therapeutic strategy and long-term disease course in a prospective referral cohort of adult IBD patients.METHODS Sera of 257 Crohn’s disease(CD)and 180 ulcerative colitis(UC)patients from a tertiary IBD referral center of Hungary(Division of Gastroenterology,Department of Internal Medicine,Faculty of Medicine,University of Debrecen)were assayed for IgG,IgA,and secretory IgA(sIgA)type aCHI3L1 by enzyme-linked immunosorbent assay using recombinant CHI3L1,along with 86 healthy controls(HCONT).RESULTS The IgA type was more prevalent in CD than in UC(29.2%vs 11.1%)or HCONT(2.83%;P<0.0001 for both).However,sIgA subtype aCHI3L1 positivity was higher in both CD and UC patients than in HCONT(39.3%and 32.8%vs 4.65%,respectively;P<0.0001).The presence of both IgA and sIgA aCHI3L1 antibodies was associated with colonic involvement(P<0.0001 and P=0.038,respectively)in patients with CD.Complicated disease behavior at sample procurement was associated with aCHI3L1 sIgA positivity(57.1%vs 36.0%,P=0.009).IgA type aCH3L1 was more prevalent in patients with frequent relapse during the disease course in the CD group(46.9%vs 25.7%,P=0.005).In a group of patients with concomitant presence of pure inflammatory luminal disease and colon involvement at the time of diagnosis,positivity for IgA or sIgA type aCH3L1 predicted faster progression towards a complicated disease course in time-dependent models.This association disappeared after merging subgroups of different disease locations.CONCLUSION CHI3L1 is a novel neutrophil autoantigenic target in IBD.The consideration of antibody classes along with location-based prediction may transform the future of serology in IBD.展开更多
BACKGROUND In recent years,the emergence of multiplex technology that can simultaneously measure multiple anti-islet autoantibodies has become particularly valuable for the staging and early diagnosis of immune-mediat...BACKGROUND In recent years,the emergence of multiplex technology that can simultaneously measure multiple anti-islet autoantibodies has become particularly valuable for the staging and early diagnosis of immune-mediated type 1 diabetes(T1D).While it has been established that 20%-30%of T1D patients suffer from autoimmune thyroid disease(AITD),there is limited available data regarding the presence of anti-islet autoantibodies in AITD patients.Among commercially available anti-islet autoantibodies,glutamic acid decarboxylase 65 autoantibodies(GADAs)are often the first marker measured in general clinical practice.AIM To investigate the frequency of anti-islet autoantibodies in AITD patients.METHODS Our study involved four hundred ninety-five AITD patients,categorized into three distinct groups:AITD with T1D(n=18),AITD with phenotypic type 2 diabetes(T2D)(n=81),and AITD without diabetes(n=396),and the enzyme-linked immunosorbent assay(ELISA)was employed to determine the frequencies of 3 Screen Islet Cell Autoantibody(3 Screen ICA),GADA,insulinoma-associated antigen-2 autoantibodies(IA-2As),and zinc transporter 8 autoantibodies(ZnT8As)within these groups.RESULTS The frequency of 3 Screen ICA in AITD patients with T1D,T2D,and those without diabetes were 88.9%,6.2%,and 5.1%,respectively,with no significant difference seen between the latter two groups.Notably,the frequency of 3 Screen ICA was 11.1%higher in AITD patients with T1D,1.3%higher in AITD patients with T2D,and 1.1%higher in AITD patients without diabetes compared to GADA,respectively.Furthermore,12.5%,20.0%,and 20.0%of the 3 Screen ICA-positive patients were negative for GADA.Additionally,1.3%of the AITD patients who tested negative for 3 Screen ICA in both the AITD with T2D and non-diabetic AITD groups were found to be positive for individual autoantibodies.Among the 3 Screen ICA-positive patients,there was a significantly higher proportion of individuals with multiple autoantibodies in AITD patients with T1D compared to those without diabetes(37.5%vs 5.0%,P<0.05).However,this proportion was similar to that in AITD patients with T2D(20.0%).Nevertheless,there was no significant difference in 3 Screen ICA titers between AITD patients with T1D and those without diabetes(436.8±66.4 vs 308.1±66.4 index).Additionally,no significant difference in 3 Screen ICA titers was observed between Graves’disease and Hashimoto’s thyroiditis in any of the groups.CONCLUSION Our findings reveal that some AITD patients without diabetes exhibit 3 Screen ICA titers comparable to those in AITD patients with T1D.Thus,3 Screen ICA outperforms GADA in identifying latent anti-islet autoantibody-positive individuals among AITD patients.展开更多
文摘目的:探讨血清基质金属蛋白酶-1(matrix metalloproteinase-1,MMP1)和纽约食管鳞状细胞癌抗原-1(New York esophageal squamous cell carcinoma 1,NY-ESO-1)自身抗体联合检测在食管鳞状细胞癌中的诊断意义。方法:应用酶联免疫吸附实验检测120例食管鳞状细胞癌患者和120例正常对照血清中MMP1和NY-ESO-1自身抗体的表达水平,采用受试者工作特征(receiver operating characteristic,ROC)曲线评价诊断效能。结果:血清MMP1和NY-ESO-1自身抗体在食管鳞状细胞癌患者中的表达均明显高于正常对照[(8.070±5.738)ng/mL vs(4.331±3.137)ng/mL,Z=6.214,P<0.001;0.463±0.571 vs 0.156±0.086,Z=5.210,P<0.001]。ROC曲线显示,当血清MMP1为最佳诊断临界值10.586 ng/mL时,其在诊断食管鳞状细胞癌的曲线下面积(area under the ROC curve,AUC)为0.732(95%CI:0.671~0.787),敏感度为24.2%,特异度为95.0%。NY-ESO-1自身抗体诊断食管鳞状细胞癌AUC为0.695(95%CI:0.632~0.752),敏感度为33.0%,特异度为95.0%。MMP1和NY-ESO-1自身抗体联合检测诊断食管鳞状细胞癌的AUC为0.800(95%CI:0.744~0.849),敏感度为47.5%,特异度为95.0%。结论:血清MMP1和NY-ESO-1自身抗体联合检测可能有助于提高食管鳞状细胞癌的诊断效能。
基金Supported by National Natural Science Foundation of China,No.81472213the Health Commission of Zhejiang Province,No.2019ZD010 and No.2019ZD029+1 种基金the Science Technology Department of Zhejiang Province,No.LGF20H220001the Zhejiang Provincial Administration of Traditional Chinese Medicine,No.2021ZA088.
文摘BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),the mechanisms are largely unknown.AIM To explore the role of BMI-1,a polycomb group protein family member,which is highly expressed in LM,and the interaction between aHSCs and CRC cells in promoting CRC liver metastasis(CRLM).METHODS Immunohistochemistry was carried out to examine BMI-1 expression in LM and matched liver specimens of CRC.The expression levels of BMI-1 in mouse liver during CRLM(0,7,14,21,and 28 d)were detected by Western blotting(WB)and the quantitative polymerase chain reaction(qPCR)assay.We overexpressed BMI-1 in HSCs(LX2)by lentivirus infection and tested the molecular markers of aHSCs by WB,qPCR,and the immunofluorescence assay.CRC cells(HCT116 and DLD1)were cultured in HSC-conditioned medium(LX2 NC CM or LX2 BMI-1 CM).CM-induced CRC cell proliferation,migration,epithelial-mesenchymal transition(EMT)phenotype,and transforming growth factor beta(TGF-β)/SMAD pathway changes were investigated in vitro.A mouse subcutaneous xenotransplantation tumor model was established by co-implantation of HSCs(LX2 NC or LX2 BMI-1)and CRC cells to investigate the effects of HSCs on tumor growth and the EMT phenotype in vivo.RESULTS Positive of BMI-1 expression in the liver of CRLM patients was 77.8%.The expression level of BMI-1 continued to increase during CRLM in mouse liver cells.LX2 overexpressed BMI-1 was activated,accompanied by increased expression level of alpha smooth muscle actin,fibronectin,TGF-β1,matrix metalloproteinases,and interleukin 6.CRC cells cultured in BMI-1 CM exhibited enhanced proliferation and migration ability,EMT phenotype and activation of the TGF-β/SMAD pathway.In addition,the TGF-βR inhibitor SB-505124 diminished the effect of BMI-1 CM on SMAD2/3 phosphorylation in CRC cells.Furthermore,BMI-1 overexpressed LX2 HSCs promoted tumor growth and the EMT phenotype in vivo.CONCLUSION High expression of BMI-1 in liver cells is associated with CRLM progression.BMI-1 activates HSCs to secrete factors to form a prometastatic environment in the liver,and aHSCs promote proliferation,migration,and the EMT in CRC cells partially through the TGF-β/SMAD pathway.
文摘目的:研究微小RNA-218(microRNA-218,miR-218)靶向B细胞特性莫洛尼鼠白血病病毒插入位点1(B-cell specific moloney leukemia virus insertion site 1,Bmi-1)抑制肺腺癌细胞增殖的作用及机制。方法:培养肺腺癌细胞株A549,分别转染miR-阴性对照(NC)核苷酸序列、miR-218核苷酸序列、NC质粒、Bmi-1质粒,采用平板克隆形成实验、CCK8实验检测细胞增殖水平,采用荧光定量PCR实验检测miR-218的表达,采用Western blot实验检测Bmi-1的表达,采用双荧光素酶报告基因实验检测miR-218与Bmi-1的靶向关系。建立移植瘤裸鼠,慢病毒转染miR-NC或miR-218核苷酸序列,检测移植瘤体积、质量及Bmi-1的表达。结果:与转染miR-NC核苷酸序列的miR-NC组比较,转染miR-218核苷酸序列的miR-218组A549细胞miR-218表达水平、增殖抑制率增加,克隆形成率、Bmi-1表达水平及野生型Bmi-1质粒的荧光素酶活性降低(P<0.05);与共转染miR-218核苷酸序列及NC质粒的miR-218+NC质粒组比较,共转染miR-218核苷酸序列及Bmi-1质粒的miR-218+Bmi-1质粒组A549细胞增殖抑制率降低,克隆形成率、Bmi-1表达水平增加(P<0.05);与慢病毒转染miR-NC核苷酸序列的miR-NC组比较,慢病毒转染miR-218核苷酸序列的miR-218组裸鼠移植瘤体积、质量及Bmi-1表达水平均降低(P<0.05)。结论:miR-218靶向下调Bmi-1表达并抑制肺腺癌细胞增殖。
基金Supported by the German Federal Ministry of Education and Research(BMBF-Wachstumskern-PRAEMED.BIO),03WKDB2Csupported by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences,BO/00232/17/5+1 种基金Research Grants of National Research Development and Innovation Office,K115818/2015/1New National Excellence Program of the Ministry of Human Capacities,ÚNKP-18-4 Bolyai Plus.
文摘BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by the infiltration of neutrophils in the inflamed mucosa and the accumulation of pathogens.Like neutrophils in the context of innate immune responses,immunoglobulin A(IgA)as an acquired immune response partakes in the defense of the intestinal epithelium.Under normal conditions,IgA contributes to the elimination of microbes,but in connection with the loss of tolerance to chitinase 3-like 1(CHI3L1)in IBD,IgA could participate in CHI3L1-mediated improved adhesion and invasion of potentially pathogenic microorganisms.The tolerance brake to CHI3L1 and the occurrence of IgA autoantibodies to this particular target,the exact role and underlying mechanisms of CHI3L1 in the pathogenesis of IBD are still unclear.AIM To determine the predictive potential of Ig subtypes of a novel serological marker,anti-CHI3L1 autoantibodies(aCHI3L1)in determining the disease phenotype,therapeutic strategy and long-term disease course in a prospective referral cohort of adult IBD patients.METHODS Sera of 257 Crohn’s disease(CD)and 180 ulcerative colitis(UC)patients from a tertiary IBD referral center of Hungary(Division of Gastroenterology,Department of Internal Medicine,Faculty of Medicine,University of Debrecen)were assayed for IgG,IgA,and secretory IgA(sIgA)type aCHI3L1 by enzyme-linked immunosorbent assay using recombinant CHI3L1,along with 86 healthy controls(HCONT).RESULTS The IgA type was more prevalent in CD than in UC(29.2%vs 11.1%)or HCONT(2.83%;P<0.0001 for both).However,sIgA subtype aCHI3L1 positivity was higher in both CD and UC patients than in HCONT(39.3%and 32.8%vs 4.65%,respectively;P<0.0001).The presence of both IgA and sIgA aCHI3L1 antibodies was associated with colonic involvement(P<0.0001 and P=0.038,respectively)in patients with CD.Complicated disease behavior at sample procurement was associated with aCHI3L1 sIgA positivity(57.1%vs 36.0%,P=0.009).IgA type aCH3L1 was more prevalent in patients with frequent relapse during the disease course in the CD group(46.9%vs 25.7%,P=0.005).In a group of patients with concomitant presence of pure inflammatory luminal disease and colon involvement at the time of diagnosis,positivity for IgA or sIgA type aCH3L1 predicted faster progression towards a complicated disease course in time-dependent models.This association disappeared after merging subgroups of different disease locations.CONCLUSION CHI3L1 is a novel neutrophil autoantigenic target in IBD.The consideration of antibody classes along with location-based prediction may transform the future of serology in IBD.
文摘BACKGROUND In recent years,the emergence of multiplex technology that can simultaneously measure multiple anti-islet autoantibodies has become particularly valuable for the staging and early diagnosis of immune-mediated type 1 diabetes(T1D).While it has been established that 20%-30%of T1D patients suffer from autoimmune thyroid disease(AITD),there is limited available data regarding the presence of anti-islet autoantibodies in AITD patients.Among commercially available anti-islet autoantibodies,glutamic acid decarboxylase 65 autoantibodies(GADAs)are often the first marker measured in general clinical practice.AIM To investigate the frequency of anti-islet autoantibodies in AITD patients.METHODS Our study involved four hundred ninety-five AITD patients,categorized into three distinct groups:AITD with T1D(n=18),AITD with phenotypic type 2 diabetes(T2D)(n=81),and AITD without diabetes(n=396),and the enzyme-linked immunosorbent assay(ELISA)was employed to determine the frequencies of 3 Screen Islet Cell Autoantibody(3 Screen ICA),GADA,insulinoma-associated antigen-2 autoantibodies(IA-2As),and zinc transporter 8 autoantibodies(ZnT8As)within these groups.RESULTS The frequency of 3 Screen ICA in AITD patients with T1D,T2D,and those without diabetes were 88.9%,6.2%,and 5.1%,respectively,with no significant difference seen between the latter two groups.Notably,the frequency of 3 Screen ICA was 11.1%higher in AITD patients with T1D,1.3%higher in AITD patients with T2D,and 1.1%higher in AITD patients without diabetes compared to GADA,respectively.Furthermore,12.5%,20.0%,and 20.0%of the 3 Screen ICA-positive patients were negative for GADA.Additionally,1.3%of the AITD patients who tested negative for 3 Screen ICA in both the AITD with T2D and non-diabetic AITD groups were found to be positive for individual autoantibodies.Among the 3 Screen ICA-positive patients,there was a significantly higher proportion of individuals with multiple autoantibodies in AITD patients with T1D compared to those without diabetes(37.5%vs 5.0%,P<0.05).However,this proportion was similar to that in AITD patients with T2D(20.0%).Nevertheless,there was no significant difference in 3 Screen ICA titers between AITD patients with T1D and those without diabetes(436.8±66.4 vs 308.1±66.4 index).Additionally,no significant difference in 3 Screen ICA titers was observed between Graves’disease and Hashimoto’s thyroiditis in any of the groups.CONCLUSION Our findings reveal that some AITD patients without diabetes exhibit 3 Screen ICA titers comparable to those in AITD patients with T1D.Thus,3 Screen ICA outperforms GADA in identifying latent anti-islet autoantibody-positive individuals among AITD patients.
