The glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney that causes kidney diseases. The reason of glomerulonephritis disease is to deposit the anti-GBM auto a...The glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney that causes kidney diseases. The reason of glomerulonephritis disease is to deposit the anti-GBM auto antibody in the glomerular basement membrane. The type IV collagen is the main component of glomerular basement membrane that has α3 chain of type (IV) collagen of non-collagenous domain which contains N-terminal 7S domain, a triple helical collagenous domain and C-terminal non-collagenous glomerular domain (NC1). The amino terminal of α3 (IV) NC1 that induces the Experimental Autoimmuno Glomerulonephritis (EAG) in rat model has been identified. The recombinant rat α3 (IV) NC1 antigen has nine amino acid spans that are consistent with antibody or T cell epitope that induces in EAG. The research is carried out on the recombinant rat α3 (IV) NC1 production, purification, quantification, and characterization. The circulation of anti-GBM antibody in glomerular basement membrane can be measured by the ELISA assay. In addition, the recombinant rat antigen is secreted in HEK293 cell supernatant that is purified by Anti-FLAG M2 monoclonal IgG antibody affinity column and characterized and quantified by SDS-PAGE gel electrophoresis and Western blotting techniques.展开更多
Aims: We evaluated whether urinary excretion of type IV collagen (U-COL) may predict an increase in the urinary albumin-to-creatinine ratio (ACR) and what factors regulate U-COL in 145 normoalbuminuric patients with t...Aims: We evaluated whether urinary excretion of type IV collagen (U-COL) may predict an increase in the urinary albumin-to-creatinine ratio (ACR) and what factors regulate U-COL in 145 normoalbuminuric patients with type 2 diabetes. Methods: We measured HbA1c, systolic blood pressure (SBP), urinary 8-hydroxydeoxyguanosine (8-OHdG) and monocyte chemoattractant protein (MCP)-1 at start of this study (Baseline), ACR and U-COL in addition to these measurements at one year later (Evaluation-1), and ACR and SBP after two years of the Evaluation-1 (Evaluation-2). The relationships were investigated between the increase of ACR and the U-COL. The effect of angiotensin receptor blockers (ARB) treatment on the correlations between U-COL and ACR at Evaluation-2 on one hand, and between U-COL and percent change of ACR on the other, was also analyzed. Furthermore, we investigated whether the increase in 8-OHdG and in MCP-1 in a year prior to the Evaluation-1 were risk factors of the rise in U-COL levels. Results: Both U-COL and SBP at Evaluation-1, but not ARB treatment, were independent risk factors for an increased ACR after 2 years. ARB treatment significantly suppressed the increase in ACR after 2 years in patients with higher U-COL excretion. The percentage changes in 8-OHdG (%8-OHdG) and MCP-1 (%MCP-1) in one year prior to Evaluation-1 measurements are independent risk factors for U-COL. HbA1c and SBP values one year prior to Evaluation-1 are independent risk factors not only for %8-OHdG but also, for baseline U-COL. The %8-OHdG is an independent risk factor for %MCP-1. Conclusions: U-COL may predict an increase in the ACR. The U-COL seems to be increased with oxidative stress and inflammation induced by past hyperglycemia.展开更多
In this work, the influence of different substrate adhesion during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of THP-1 monocytic cell line was studied. In particular, by morphocytochemical and cytom...In this work, the influence of different substrate adhesion during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of THP-1 monocytic cell line was studied. In particular, by morphocytochemical and cytometric approaches, the influence of type I and type IV collagens in an experimental model representative of three phases (initial, intermediate and terminal) of monocyte-macrophage transition was analyzed. The cells in these three phases of differentiation were obtained by using 6, 30 e 60 nM PMA. In this experimental model, referring to adhesion to glass as control, by using the azo-dye coupling method, we have considered the analysis of Acid Phosphatase (AcP) activity as a marker of differentiated status expression, in relation to the acquisition of macrophagic phenotype. Endosomal/lysosomal system was further characterized by taking into account the uptake of fluorescent probe LysoTracker Red. Fluorochromization in the various experimental conditions was analyzed morphologically (fluorescence microscopy) and quantitatively (static cytometry). Data related to lysosome compartment were integrated, from a cytokinetic point of view, by flow cytometry measurements of DNA/protein content. Our results have indicated that type I and type IV collagens were able to influence, with respect to glass adhesion, various differentiation phases. Type I collagen showed the higher effects in the condition of high differentiation (60 nM PMA), causing an increase in AcP activity and lysosomal system. Type IV collagen, besides determining effects on lysosomal compartment of intermediate and terminally differentiated cells, influenced mainly proliferative activity of cells with initial differentiation level (6 nM PMA).展开更多
Cancer cell metabolism reprogramming is one of the hallmarks of cancer.Cancer cells preferentially utilize aerobic glycolysis,which is regulated by activated oncogenes and the tumor microenvironment.Extracellular matr...Cancer cell metabolism reprogramming is one of the hallmarks of cancer.Cancer cells preferentially utilize aerobic glycolysis,which is regulated by activated oncogenes and the tumor microenvironment.Extracellular matrix(ECM)in the tumor microenvironment,including the basement membranes(BMs),is dynamically remodeled.However,whether and how ECM regulates tumor glycolysis is largely unknown.We show that type IV collagens,components of BMs essential for the tissue integrity and proper function,are differentially expressed in breast cancer subtypes thatα5 chain(α5(IV))is preferentially expressed in the luminal-type breast cancer and is regulated by estrogen receptor-α.α5(IV)is indispensable for luminal breast cancer development.Ablation ofα5(IV)significantly reduces the growth of luminal-type breast cancer cells and impedes the development of luminal-type breast cancer.Impaired cell growth and tumor development capability ofα5(IV)-ablated luminal breast cancer cells is attributed to the reduced expression of glucose transporter and glycolytic enzymes and impaired glycolysis in luminal breast cancer cells.Non-integrin collagen receptor discoidin domain receptor-1(DDR1)expression and p38 mitogen-activated protein kinase activation are attenuated inα5(IV)-ablated luminal breast cancer cells,resulting in reduced c-Myc oncogene expression and phosphorylation.Ectopic expression of constitutively active DDR1 or c-Myc restores the expression of glucose transporter and glycolytic enzymes,and thereafter restores aerobic glycolysis,cell proliferation,and tumor growth of luminal breast cancer.Thus,type IV collagenα5 chain is a luminal-type breast cancer-specific microenvironmental regulator modulating cancer cell metabolism.展开更多
BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced li...BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced liver fibrosis is known to be poor.It is difficult to assess disease progression in all patients with NAFLD;thus,it is necessary to identify patients who will show poor prognosis.AIM To investigate the efficacy of non-invasive biomarkers for predicting disease progression in patients with NAFLD.METHODS We investigated biomarkers associated with mortality in patients with NAFLD who visited the Kawasaki Medical School General Medical Center from 1996 to 2018 and underwent liver biopsy and had been followed-up for>1 year.Cumulative overall mortality and liver-related events during follow-up were calculated using the Kaplan-Meier analysis and compared using log-rank testing.We calculated the odds ratio and performed receiver operating characteristic curve analysis with logistic regression analysis to determine the optimal cut-off value with the highest prognostic ability.RESULTS We enrolled 489 patients who were followed-up for a period of 1-22.2 years.In total,13 patients died(2.7%of total patients enrolled);7 patients died due to liverrelated causes.Poor prognosis was associated with liver fibrosis on histological examination but not with inflammation or steatosis.Blood biomarkers associated with mortality were platelet counts,albumin levels,and type IV collagen 7S levels.The optimal cutoff index for predicting total mortality was a platelet count of 15×10^(4)/μL,albumin level of 3.5 g/dL,and type IV collagen 7S level of 5 mg/dL.In particular,only one-factor patients with NAFLD presenting with platelet counts≤15×10^(4)/μL,albumin levels≤3.5 g/dL,or type IV collagen 7S≥5 mg/dL showed 5-year,10-year,and 15-year survival rates of 99.7%,98.3%,and 94%,respectively.However,patients with two factors had lower 5-year and 10-year survival rates of 98%and 43%,respectively.Similarly,patients with all three factors showed the lowest 5-year and 10-year survival rates of 53%and 26%,respectively.CONCLUSION A combination of the three non-invasive biomarkers is a useful predictor of NAFLD prognosis and can help identify patients with NAFLD who are at a high risk of all-cause mortality.展开更多
BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments o...BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.展开更多
Some organic phosphines labeled with ^117mSn(IV) have been proved promising for imaging and pain palliation of bone tumor.In this paper,a preliminary investigation on the adsorption characteristics of EDTMP(ethelenedi...Some organic phosphines labeled with ^117mSn(IV) have been proved promising for imaging and pain palliation of bone tumor.In this paper,a preliminary investigation on the adsorption characteristics of EDTMP(ethelenediaminetetramethylene phosphoric acid)labeled with ^117Sn(IV) on HA( hydroxyapatitie)and collagen,and an investigation on the adsorption mechanism of ^117mSn(IV)-EDTMP on HA was presented.展开更多
文摘The glomerulonephritis disease is characterized by inflammation of glomeruli or small blood vessels in the kidney that causes kidney diseases. The reason of glomerulonephritis disease is to deposit the anti-GBM auto antibody in the glomerular basement membrane. The type IV collagen is the main component of glomerular basement membrane that has α3 chain of type (IV) collagen of non-collagenous domain which contains N-terminal 7S domain, a triple helical collagenous domain and C-terminal non-collagenous glomerular domain (NC1). The amino terminal of α3 (IV) NC1 that induces the Experimental Autoimmuno Glomerulonephritis (EAG) in rat model has been identified. The recombinant rat α3 (IV) NC1 antigen has nine amino acid spans that are consistent with antibody or T cell epitope that induces in EAG. The research is carried out on the recombinant rat α3 (IV) NC1 production, purification, quantification, and characterization. The circulation of anti-GBM antibody in glomerular basement membrane can be measured by the ELISA assay. In addition, the recombinant rat antigen is secreted in HEK293 cell supernatant that is purified by Anti-FLAG M2 monoclonal IgG antibody affinity column and characterized and quantified by SDS-PAGE gel electrophoresis and Western blotting techniques.
文摘Aims: We evaluated whether urinary excretion of type IV collagen (U-COL) may predict an increase in the urinary albumin-to-creatinine ratio (ACR) and what factors regulate U-COL in 145 normoalbuminuric patients with type 2 diabetes. Methods: We measured HbA1c, systolic blood pressure (SBP), urinary 8-hydroxydeoxyguanosine (8-OHdG) and monocyte chemoattractant protein (MCP)-1 at start of this study (Baseline), ACR and U-COL in addition to these measurements at one year later (Evaluation-1), and ACR and SBP after two years of the Evaluation-1 (Evaluation-2). The relationships were investigated between the increase of ACR and the U-COL. The effect of angiotensin receptor blockers (ARB) treatment on the correlations between U-COL and ACR at Evaluation-2 on one hand, and between U-COL and percent change of ACR on the other, was also analyzed. Furthermore, we investigated whether the increase in 8-OHdG and in MCP-1 in a year prior to the Evaluation-1 were risk factors of the rise in U-COL levels. Results: Both U-COL and SBP at Evaluation-1, but not ARB treatment, were independent risk factors for an increased ACR after 2 years. ARB treatment significantly suppressed the increase in ACR after 2 years in patients with higher U-COL excretion. The percentage changes in 8-OHdG (%8-OHdG) and MCP-1 (%MCP-1) in one year prior to Evaluation-1 measurements are independent risk factors for U-COL. HbA1c and SBP values one year prior to Evaluation-1 are independent risk factors not only for %8-OHdG but also, for baseline U-COL. The %8-OHdG is an independent risk factor for %MCP-1. Conclusions: U-COL may predict an increase in the ACR. The U-COL seems to be increased with oxidative stress and inflammation induced by past hyperglycemia.
文摘In this work, the influence of different substrate adhesion during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of THP-1 monocytic cell line was studied. In particular, by morphocytochemical and cytometric approaches, the influence of type I and type IV collagens in an experimental model representative of three phases (initial, intermediate and terminal) of monocyte-macrophage transition was analyzed. The cells in these three phases of differentiation were obtained by using 6, 30 e 60 nM PMA. In this experimental model, referring to adhesion to glass as control, by using the azo-dye coupling method, we have considered the analysis of Acid Phosphatase (AcP) activity as a marker of differentiated status expression, in relation to the acquisition of macrophagic phenotype. Endosomal/lysosomal system was further characterized by taking into account the uptake of fluorescent probe LysoTracker Red. Fluorochromization in the various experimental conditions was analyzed morphologically (fluorescence microscopy) and quantitatively (static cytometry). Data related to lysosome compartment were integrated, from a cytokinetic point of view, by flow cytometry measurements of DNA/protein content. Our results have indicated that type I and type IV collagens were able to influence, with respect to glass adhesion, various differentiation phases. Type I collagen showed the higher effects in the condition of high differentiation (60 nM PMA), causing an increase in AcP activity and lysosomal system. Type IV collagen, besides determining effects on lysosomal compartment of intermediate and terminally differentiated cells, influenced mainly proliferative activity of cells with initial differentiation level (6 nM PMA).
基金supported by grants from the Ministry of Science and Technology of China(2020YFA0803203)the National Natural Science Foundation of China(81430067,32070789,and 31900514).
文摘Cancer cell metabolism reprogramming is one of the hallmarks of cancer.Cancer cells preferentially utilize aerobic glycolysis,which is regulated by activated oncogenes and the tumor microenvironment.Extracellular matrix(ECM)in the tumor microenvironment,including the basement membranes(BMs),is dynamically remodeled.However,whether and how ECM regulates tumor glycolysis is largely unknown.We show that type IV collagens,components of BMs essential for the tissue integrity and proper function,are differentially expressed in breast cancer subtypes thatα5 chain(α5(IV))is preferentially expressed in the luminal-type breast cancer and is regulated by estrogen receptor-α.α5(IV)is indispensable for luminal breast cancer development.Ablation ofα5(IV)significantly reduces the growth of luminal-type breast cancer cells and impedes the development of luminal-type breast cancer.Impaired cell growth and tumor development capability ofα5(IV)-ablated luminal breast cancer cells is attributed to the reduced expression of glucose transporter and glycolytic enzymes and impaired glycolysis in luminal breast cancer cells.Non-integrin collagen receptor discoidin domain receptor-1(DDR1)expression and p38 mitogen-activated protein kinase activation are attenuated inα5(IV)-ablated luminal breast cancer cells,resulting in reduced c-Myc oncogene expression and phosphorylation.Ectopic expression of constitutively active DDR1 or c-Myc restores the expression of glucose transporter and glycolytic enzymes,and thereafter restores aerobic glycolysis,cell proliferation,and tumor growth of luminal breast cancer.Thus,type IV collagenα5 chain is a luminal-type breast cancer-specific microenvironmental regulator modulating cancer cell metabolism.
文摘BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced liver fibrosis is known to be poor.It is difficult to assess disease progression in all patients with NAFLD;thus,it is necessary to identify patients who will show poor prognosis.AIM To investigate the efficacy of non-invasive biomarkers for predicting disease progression in patients with NAFLD.METHODS We investigated biomarkers associated with mortality in patients with NAFLD who visited the Kawasaki Medical School General Medical Center from 1996 to 2018 and underwent liver biopsy and had been followed-up for>1 year.Cumulative overall mortality and liver-related events during follow-up were calculated using the Kaplan-Meier analysis and compared using log-rank testing.We calculated the odds ratio and performed receiver operating characteristic curve analysis with logistic regression analysis to determine the optimal cut-off value with the highest prognostic ability.RESULTS We enrolled 489 patients who were followed-up for a period of 1-22.2 years.In total,13 patients died(2.7%of total patients enrolled);7 patients died due to liverrelated causes.Poor prognosis was associated with liver fibrosis on histological examination but not with inflammation or steatosis.Blood biomarkers associated with mortality were platelet counts,albumin levels,and type IV collagen 7S levels.The optimal cutoff index for predicting total mortality was a platelet count of 15×10^(4)/μL,albumin level of 3.5 g/dL,and type IV collagen 7S level of 5 mg/dL.In particular,only one-factor patients with NAFLD presenting with platelet counts≤15×10^(4)/μL,albumin levels≤3.5 g/dL,or type IV collagen 7S≥5 mg/dL showed 5-year,10-year,and 15-year survival rates of 99.7%,98.3%,and 94%,respectively.However,patients with two factors had lower 5-year and 10-year survival rates of 98%and 43%,respectively.Similarly,patients with all three factors showed the lowest 5-year and 10-year survival rates of 53%and 26%,respectively.CONCLUSION A combination of the three non-invasive biomarkers is a useful predictor of NAFLD prognosis and can help identify patients with NAFLD who are at a high risk of all-cause mortality.
文摘BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.
基金Supported by the Chinese Academy of Engineering Physics JM fund and Institute of Nuclear Physics and Chemistry Master fund
文摘Some organic phosphines labeled with ^117mSn(IV) have been proved promising for imaging and pain palliation of bone tumor.In this paper,a preliminary investigation on the adsorption characteristics of EDTMP(ethelenediaminetetramethylene phosphoric acid)labeled with ^117Sn(IV) on HA( hydroxyapatitie)and collagen,and an investigation on the adsorption mechanism of ^117mSn(IV)-EDTMP on HA was presented.