The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These syst...The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.展开更多
目的:探讨微环境中Ⅸ型胶原α1(collagen type IX alpha 1 chain,COL9A1)在结直肠腺癌中的表达及其与肿瘤进展的相关性和临床意义。方法:收集2012年1月至2021年1月手术切除的结直肠癌标本408例,采用免疫组织化学检测结直肠腺癌肿瘤组织...目的:探讨微环境中Ⅸ型胶原α1(collagen type IX alpha 1 chain,COL9A1)在结直肠腺癌中的表达及其与肿瘤进展的相关性和临床意义。方法:收集2012年1月至2021年1月手术切除的结直肠癌标本408例,采用免疫组织化学检测结直肠腺癌肿瘤组织及癌旁正常组织中COL9A1表达,同时检测肿瘤组织中肿瘤蛋白53(tumor protein 53,P53)和错配修复(mismatch repair,MMR)蛋白MLH1、MSH6和PMS2的表达,统计分析COL9A1的表达与各临床病理特征参数的关系,以及与P53突变和MMR状态的相关性,并分析COL9A1阳性表达患者的预后情况。结果:COL9A1在结直肠腺癌肿瘤组织中表达显著低于癌旁正常组织(P<0.001);COL9A1的表达与肿瘤浸润深度、临床分期和肠系膜淋巴结转移有关(χ^(2)=16.943、89.031和84.814;均P<0.001),而与P53突变和MMR状态无关(χ^(2)=0.677、1.260,均P>0.05);Log-rank检验显示COL9A1阴性表达患者的无进展生存期(progression free survival,PFS)和总体生存期(overall survival,OS)显著低于COL9A1阳性表达患者(分别P<0.001,P=0.040)。结论:结直肠腺癌中COL9A1蛋白的表达缺失与肿瘤浸润及转移密切相关,并提示不良预后,这可为结直肠癌预后评估、药物筛选等提供可能的分子标志物和治疗策略。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
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
AIM: To investigate the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen I synthesis of human keratocyte on titanium(Ti) surface. · METHODS: The chimeric peptide RKLPDAPRGDN(minTBP-1-PRGDN...AIM: To investigate the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen I synthesis of human keratocyte on titanium(Ti) surface. · METHODS: The chimeric peptide RKLPDAPRGDN(minTBP-1-PRGDN) was synthesized by connecting RKLPDA(minTBP-1) to the N-terminal of PRGDN,the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen I synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1 as controls. The keratocytes attached to the surface of Ti were either stained with FITC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim-ethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide uptaking methods. The secretion of type I collagen was determined using an ELISA kit. ·RESULTS: The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti(1.40±0.03 folds,P =0.003),to promote the proliferation(1.26 ±0.05 folds,P =0.014) and the synthesis of type I collagen(1.530 ±0.128,P =0.008). MinTBP-1 at the same concentration could only promote the attachment(1.13±0.04 folds,P =0.020) and proliferation(1.15±0.06 folds,P =0.021),while PRGDN had no significant influence(P 】0.05). ·CONCLUSION: Our data show that the novel chimericpeptide minTBP-1-PRGDN could promote the attachment,proliferation and type I collagen synthesis of human keratocytes on the surface of titanium.展开更多
AIM: To investigate the interfering effect of Y-27632, a ROCK-I selective inhibitor, on the signal transduction pathway of transforming growth factor-beta 1 (TGF-beta 1) in ocular Tenon capsule fibroblasts (OTFS) in v...AIM: To investigate the interfering effect of Y-27632, a ROCK-I selective inhibitor, on the signal transduction pathway of transforming growth factor-beta 1 (TGF-beta 1) in ocular Tenon capsule fibroblasts (OTFS) in vitro. METHODS: After OTFS from passages 4 to 6 47 vitro were induced by TGF-beta 1 and then treated by Y-27632, the changes of the OTFS cell cycles were analyzed via flow cytometry, and the proteins expression of the alpha -smooth muscular actin (alpha -SMA), connective tissue growth factor (CTGF), collagen I were calculated by Western blot. After OTFS treated by the different concentrations of Y-27632, the expression levels of the alpha -SMA, CTGF and collagen I mRNA were assayed by RT-PCR. RESULTS: Y-27632 had no markedly effect on the OTFS cell cycles. After treated by TGF-beta 1, OTFS in G1 period significantly increased. The cell cycles distribution by both TGF-beta 1 and Y-27632 had no remarkable difference from that in control group. Y-27632 significantly inhibited the proteins expressions of both alpha -SMA and CTGF, while to some extent inhibited that of collagen I. TGF-beta 1 significantly promoted the proteins expressions of alpha -SMA, CTGF and collagen I. After OTFS treated by both TGF-beta 1 and Y-27632, of alpha -SMA, the protein expression was similar with that in control group (P=0.066>0.05), but the protein expression of CTGF or collagen I, respectively, was significantly different from that in control group (P=0.000<0.01). The differences of expressions of the alpha -SMA, CTGF and collagen I mRNA in 30, 150, 750 mu mol/L Y-27632 group were statistically significant, compared with those in control group, respectively (alpha -SMA, P=0.002, 0.000, 0.000; CTGF, P=0.014, 0.002, 0.001; collagen I,P=0.003, 0.002, 0.000). CONCLUSION: Blocking the Rho/ROCK signaling pathway by using of Y-27632 could inhibit the cellular proliferation and the expression of both CTGF and alpha -SMA whatever OTFS induced by TGF-beta 1 or not. Y-27632 suppressed the expression of collagen I mRNA without induction.展开更多
目的分析北京地区健康绝经期前女性不同年龄阶段血清I型原胶原氨基端肽(procollagen type 1 N-terminal propeptide,P1NP)和I型胶原羧基端肽交联(βcross-linked C-telopeptide of type 1 collagen,β-CTX)水平的分布趋势差异并初步建...目的分析北京地区健康绝经期前女性不同年龄阶段血清I型原胶原氨基端肽(procollagen type 1 N-terminal propeptide,P1NP)和I型胶原羧基端肽交联(βcross-linked C-telopeptide of type 1 collagen,β-CTX)水平的分布趋势差异并初步建立两者的参考区间。方法以北京地区健康绝经期前女性作为研究对象,应用罗氏电化学发光免疫分析技术,对符合入组标准的272名30~54岁女性血清P1NP和β-CTX水平进行检测。以5岁为一年龄段进行分组:30~34岁,35~39岁,40~44岁,45~49岁,50~54岁;运用局部加权回归散点平滑法和Kolmogorov-Smirnov Z检验比较不同年龄段两者的组间分布趋势差异,确定参考人群的特异年龄段,并应用非参数方法建立参考区间。结果 272名入组受试者的平均年龄为(39.51±5.85)岁,总体P1NP与β-CTX水平呈非正态分布。35~39岁与40~44岁的血清P1NP与β-CTX水平分布趋势比较差异无统计学意义(P>0.05);进一步将30~34岁及45~49岁水平分别与35~44岁水平的分布趋势比较,差异有统计学意义(P<0.05)。因此将35~44岁年龄段的健康绝经期前女性作为参考人群,由此所建立的血清P1NP参考区间为:17.95~65.60 ng/m L,血清β-CTX参考区间为0.10~0.49 ng/m L。结论北京地区35~44岁健康绝经期前女性血清骨转换标志物P1NP和β-CTX水平分布趋势相对平稳,受变异因素影响最小,两者在此年龄段人群的测定结果适宜作为建立参考区间的参考值。展开更多
文摘The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.
文摘目的:探讨微环境中Ⅸ型胶原α1(collagen type IX alpha 1 chain,COL9A1)在结直肠腺癌中的表达及其与肿瘤进展的相关性和临床意义。方法:收集2012年1月至2021年1月手术切除的结直肠癌标本408例,采用免疫组织化学检测结直肠腺癌肿瘤组织及癌旁正常组织中COL9A1表达,同时检测肿瘤组织中肿瘤蛋白53(tumor protein 53,P53)和错配修复(mismatch repair,MMR)蛋白MLH1、MSH6和PMS2的表达,统计分析COL9A1的表达与各临床病理特征参数的关系,以及与P53突变和MMR状态的相关性,并分析COL9A1阳性表达患者的预后情况。结果:COL9A1在结直肠腺癌肿瘤组织中表达显著低于癌旁正常组织(P<0.001);COL9A1的表达与肿瘤浸润深度、临床分期和肠系膜淋巴结转移有关(χ^(2)=16.943、89.031和84.814;均P<0.001),而与P53突变和MMR状态无关(χ^(2)=0.677、1.260,均P>0.05);Log-rank检验显示COL9A1阴性表达患者的无进展生存期(progression free survival,PFS)和总体生存期(overall survival,OS)显著低于COL9A1阳性表达患者(分别P<0.001,P=0.040)。结论:结直肠腺癌中COL9A1蛋白的表达缺失与肿瘤浸润及转移密切相关,并提示不良预后,这可为结直肠癌预后评估、药物筛选等提供可能的分子标志物和治疗策略。
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).
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.
基金Supported by National Natural Science Foundation of China(No.81200661)Nature Science Foundation of Hubei Province(No.2010CDB09802)Wuhan Chenguang Plan Grant(No.201150431124)
文摘AIM: To investigate the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen I synthesis of human keratocyte on titanium(Ti) surface. · METHODS: The chimeric peptide RKLPDAPRGDN(minTBP-1-PRGDN) was synthesized by connecting RKLPDA(minTBP-1) to the N-terminal of PRGDN,the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen I synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1 as controls. The keratocytes attached to the surface of Ti were either stained with FITC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim-ethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide uptaking methods. The secretion of type I collagen was determined using an ELISA kit. ·RESULTS: The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti(1.40±0.03 folds,P =0.003),to promote the proliferation(1.26 ±0.05 folds,P =0.014) and the synthesis of type I collagen(1.530 ±0.128,P =0.008). MinTBP-1 at the same concentration could only promote the attachment(1.13±0.04 folds,P =0.020) and proliferation(1.15±0.06 folds,P =0.021),while PRGDN had no significant influence(P 】0.05). ·CONCLUSION: Our data show that the novel chimericpeptide minTBP-1-PRGDN could promote the attachment,proliferation and type I collagen synthesis of human keratocytes on the surface of titanium.
基金Shaanxi Province Science and Technology Gongguan Program, China (No.2011-K14-02-03)
文摘AIM: To investigate the interfering effect of Y-27632, a ROCK-I selective inhibitor, on the signal transduction pathway of transforming growth factor-beta 1 (TGF-beta 1) in ocular Tenon capsule fibroblasts (OTFS) in vitro. METHODS: After OTFS from passages 4 to 6 47 vitro were induced by TGF-beta 1 and then treated by Y-27632, the changes of the OTFS cell cycles were analyzed via flow cytometry, and the proteins expression of the alpha -smooth muscular actin (alpha -SMA), connective tissue growth factor (CTGF), collagen I were calculated by Western blot. After OTFS treated by the different concentrations of Y-27632, the expression levels of the alpha -SMA, CTGF and collagen I mRNA were assayed by RT-PCR. RESULTS: Y-27632 had no markedly effect on the OTFS cell cycles. After treated by TGF-beta 1, OTFS in G1 period significantly increased. The cell cycles distribution by both TGF-beta 1 and Y-27632 had no remarkable difference from that in control group. Y-27632 significantly inhibited the proteins expressions of both alpha -SMA and CTGF, while to some extent inhibited that of collagen I. TGF-beta 1 significantly promoted the proteins expressions of alpha -SMA, CTGF and collagen I. After OTFS treated by both TGF-beta 1 and Y-27632, of alpha -SMA, the protein expression was similar with that in control group (P=0.066>0.05), but the protein expression of CTGF or collagen I, respectively, was significantly different from that in control group (P=0.000<0.01). The differences of expressions of the alpha -SMA, CTGF and collagen I mRNA in 30, 150, 750 mu mol/L Y-27632 group were statistically significant, compared with those in control group, respectively (alpha -SMA, P=0.002, 0.000, 0.000; CTGF, P=0.014, 0.002, 0.001; collagen I,P=0.003, 0.002, 0.000). CONCLUSION: Blocking the Rho/ROCK signaling pathway by using of Y-27632 could inhibit the cellular proliferation and the expression of both CTGF and alpha -SMA whatever OTFS induced by TGF-beta 1 or not. Y-27632 suppressed the expression of collagen I mRNA without induction.
