AIM:To elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)mod...AIM:To elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)model was established before and after intravitreous injection of anti-VEGF-165b antibody.The expression of VEGF-165b and zonula occludens-1(ZO-1)in rat retina was detected by double immunofluorescence staining and Western blotting,and the breakdown of BRB was detected by Evans blue(EB)dye.RESULTS:The intact retina of rats expressed VEGF-165b and ZO-1 protein,which were mainly located in the retinal ganglion cell layer and the inner nuclear layer and were both co-expressed with vascular endothelial cell markers CD31.After acute HIOP,the expression of VEGF-165b was up-regulated;the expression of ZO-1 was down-regulated at 12h and then recovered at 3d;EB leakage increased,peaking at 12h.After intravitreous injection of anti-VEGF-165b antibody,the expression of VEGF-165b protein was no significantly changed;and the down-regulation of the expression of ZO-1 was more obvious;EB leakage became more serious,peaking at 3d.EB analysis also showed that EB leakage in the peripheral retina was greater than that in the central retina.CONCLUSION:The endogenous VEGF-165b protein may protect the BRB from acute HIOP by regulating the expression of ZO-1.The differential destruction of BRB after acute HIOP may be related to the selective loss of retinal ganglion cells.展开更多
In the early stages of traumatic spinal cord injury,extensive accumulation of autophagosomes creates a neurotoxic microenvironment,exacerbating neuronal cell death and worsening tissue damage,ultimately hindering neur...In the early stages of traumatic spinal cord injury,extensive accumulation of autophagosomes creates a neurotoxic microenvironment,exacerbating neuronal cell death and worsening tissue damage,ultimately hindering neurofunctional recovery.Activin A is a critical growth factor necessary for the development of the embryonic nervous system and for maintaining neuronal function in the adult cerebral cortex.It can inhibit excessive autophagy in ischemic stroke to reduce neuronal damage.However,the specific mechanism through which Activin A functions in the spinal cord remains poorly understood.In this study,we administered different concentrations of Activin A to neural stem cells from the spinal cord and found that Activin A stimulated the proliferation and neuronal differentiation of neural stem cells.Then,we established an in vitro oxidative stress model by using hydrogen peroxide to stimulate the neural stem cells-induced neurons.We found that Activin A could reduce apoptosis caused by oxidative stress.Subsequently,we treated a mouse model of spinal cord contusion with intrathecal injection of Activin A.Behavioral and electrophysiological results showed that Activin A promoted recovery of motor function and reconstruction of neural circuits in the model mice.Finally,RNA sequencing indicated that Activin A inhibited autophagy by activating the PI3K/AKT/mTOR pathway and upregulating the expression of synaptogenesis-related factor Sema3A in the spinal cord.These results suggest that Activin A may mediate the excessive autophagic response after spinal cord injury,promote the reconstruction of damaged neural circuits,and restore neurological function in the injured spinal cord.展开更多
Multiple sclerosis is a severe autoimmune disorder that is mainly mediated by pathogenic cluster of CD4^(+)T cell subsets.Despite advancements in the management of multiple sclerosis,there is a critical need for more ...Multiple sclerosis is a severe autoimmune disorder that is mainly mediated by pathogenic cluster of CD4^(+)T cell subsets.Despite advancements in the management of multiple sclerosis,there is a critical need for more effective and safer treatments.In the present study,we administered Lycium barbarum glycopeptide to a mouse model of experimental autoimmune encephalomyelitis-an animal model of multiple sclerosis-and evaluated its effects on pathogenic CD4^(+)T cell activation both in vivo and in vitro.Lycium barbarum glycopeptide significantly mitigated the clinical severity of experimental autoimmune encephalomyelitis,as demonstrated by reduced demyelination and neuroinflammation.Moreover,Lycium barbarum glycopeptide treatment decreased the infiltration of peripheral leukocytes into the central nervous system and suppressed pro-inflammatory cytokine expression.Lycium barbarum glycopeptide also modulated pathogenic CD4^(+)T cell activation by inhibiting T helper 1/T helper 17 cell differentiation while promoting regulatory T cell expansion.Notably,no side effects were observed,suggesting the long-term safety and tolerability of Lycium barbarum glycopeptide.Furthermore,RNA sequencing data indicated that Lycium barbarum glycopeptide inhibits activator protein-1,an essential regulator of T cell activation and differentiation.This finding was supported by the reversal of T helper/T helper 17 cell response suppression upon AP-1 blockade.Collectively,these results highlight the potential of Lycium barbarum glycopeptide as an innovative therapeutic agent for CD4^(+)T cell-associated autoimmune or inflammatory diseases,such as multiple sclerosis.展开更多
目的研究VEGF165b和VEGF165在胃癌及癌旁正常组织中的表达及胃癌组织中VEGF165b m RNA/VEGF165 m RNA与预后的关系。方法分别应用实时RT-PCR和免疫组织化学法检测胃癌及癌旁正常组织中VEGF165b和VEGF165的基因及蛋白表达,并对患者进行2...目的研究VEGF165b和VEGF165在胃癌及癌旁正常组织中的表达及胃癌组织中VEGF165b m RNA/VEGF165 m RNA与预后的关系。方法分别应用实时RT-PCR和免疫组织化学法检测胃癌及癌旁正常组织中VEGF165b和VEGF165的基因及蛋白表达,并对患者进行2年的随访,比较VEGF165b m RNA/VEGF165 m RNA比值及这两种蛋白表达水平在胃癌及癌旁组织中的变化;同时分析VEGF165b与VEGF165蛋白表达的相关性以及胃癌组织中VEGF165b m RNA/VEGF165 m RNA与预后的关系。结果在胃癌组织中VEGF165b m RNA/VEGF165 m RNA低于癌旁正常组织中的水平;在癌旁正常组织中VEGF165b基因及蛋白表达强于VEGF165,在胃癌组织中VEGF165b基因及蛋白的表达则低于VEGF165,而且两者表达呈负相关关系;此外,在两年内死亡的患者中VEGF165b m RNA/VEGF165 m RNA明显低于生存患者中的比值水平。结论与正常组织相比,胃癌组织中存在VEGF165b向VEGF165基因及蛋白水平上的优势转换,这种变化有望成为治疗胃癌的新靶点。展开更多
目的:对比观察VEGF165b重组蛋白(recombinant human VEGF165b protein,rhVEGF165b)与贝伐珠单抗对人胃癌裸鼠移植瘤生长的影响及作用机制.方法:用人胃癌细胞BGC-823接种于裸鼠皮下,建立裸鼠移植瘤模型,随机分为3组,每组21只:rhVEGF165b...目的:对比观察VEGF165b重组蛋白(recombinant human VEGF165b protein,rhVEGF165b)与贝伐珠单抗对人胃癌裸鼠移植瘤生长的影响及作用机制.方法:用人胃癌细胞BGC-823接种于裸鼠皮下,建立裸鼠移植瘤模型,随机分为3组,每组21只:rhVEGF165b组(腹腔注射rhVEGF165b 10?g/kg)、贝伐珠单抗组(腹腔注射贝伐珠单抗5 mg/kg)、对照组(腹腔注射生理盐水10 mL/kg),分别于1、2、3 wk测量裸鼠移植瘤体积及瘤质量,免疫组织化学方法检测肿瘤组织中CD34表达(以阳性细胞数计算肿瘤微血管密度),TUNEL方法检测肿瘤组织细胞凋亡.结果:贝伐珠单抗组移植瘤体积(cm3)及瘤质量(g)在第1、2、3周均小于对照组(1 wk:0.453±0.119 vs 0.637±0.084,0.32±0.097vs 0.46±0.093;2 wk:1.691±0.381 vs 2.238±0.29,1.168±0.524 vs 2.013±0.833;3 wk:1.709±0.474 vs 4.872±0.594,1.747±0.557 vs3.463±0.986,均P<0.05),而rhVEGF165b组仅在第1、2周较对照组小(1 wk:0.546±0.132 vs0.637±0.084,1.894±0.599 vs 0.46±0.093;2wk:1.894±0.599 vs 2.238±0.29,1.537±0.568vs 2.013±0.833,均P<0.05),第3周其体积及瘤质量大于贝伐珠单抗组(3 wk:3.843±1.339 vs1.709±0.474,3.066±1.281 vs 1.747±0.557,P<0.05),与对照组无明显差异.rhVEGF165b组、贝伐珠单抗组CD34的表达水平在第1、2、3周均低于对照组(P<0.05,P<0.01),凋亡指数高于对照组(P<0.05,P<0.01);与贝伐珠单抗组相比,第1周rhVEGF165b组CD34的表达水平更低(P<0.05),但第2、3周却高于贝伐珠单抗组(P<0.01);rhVEGF165b组的凋亡指数在第1周明显高于贝伐珠单抗组(P<0.01),而第2、3周却低于贝伐珠单抗组(P<0.05).结论:rhVEGF165b、贝伐珠单抗对人胃癌移植瘤的生长有明显抑制作用,rhVEGF165b早期抑制血管生成、促进细胞凋亡表现更为明显.展开更多
目的:初步探讨VEGF165b与膀胱移行细胞癌(Transitional cell carcinoma of the bladder,TCCB)发生发展的关系。方法:用S-P免疫组化法检测38例TCCB组织和36例正常膀胱组织中VEGF165b蛋白表达;以RT-PCR法检测55例TCCB组织和43例正常膀胱...目的:初步探讨VEGF165b与膀胱移行细胞癌(Transitional cell carcinoma of the bladder,TCCB)发生发展的关系。方法:用S-P免疫组化法检测38例TCCB组织和36例正常膀胱组织中VEGF165b蛋白表达;以RT-PCR法检测55例TCCB组织和43例正常膀胱组织中VEGF165bmRNA表达。结果:正常膀胱组织中VEGF165b蛋白表达率为97.22%(35/36),TCCB组织中VEGF165b蛋白表达率为21.05%(8/38),表达差异有显著性(P<0.05);正常膀胱组织中VEGF165bmRNA阳性表达率为95.35%(41/43),明显高于TCCB组织中VEGF165bmRNA阳性表达率18.18%(10/55)(P<0.05);TCCB中VEGF165b蛋白和mRNA的表达与TCCB不同分期、分级间的表达差异具有统计学意义(P<0.05)。结论:VEGF165b在TCCB组织中的表达明显低于正常膀胱组织中的表达,这种表达差异可能与VEGF165b抑制膀胱癌的发生、发展作用有关。展开更多
基金Supported by the National Natural Science Foundation of China(No.81660217)Youth Foundation of the First Affiliated Hospital of Hainan Medical University(No.HYYFYPY201922)。
文摘AIM:To elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)model was established before and after intravitreous injection of anti-VEGF-165b antibody.The expression of VEGF-165b and zonula occludens-1(ZO-1)in rat retina was detected by double immunofluorescence staining and Western blotting,and the breakdown of BRB was detected by Evans blue(EB)dye.RESULTS:The intact retina of rats expressed VEGF-165b and ZO-1 protein,which were mainly located in the retinal ganglion cell layer and the inner nuclear layer and were both co-expressed with vascular endothelial cell markers CD31.After acute HIOP,the expression of VEGF-165b was up-regulated;the expression of ZO-1 was down-regulated at 12h and then recovered at 3d;EB leakage increased,peaking at 12h.After intravitreous injection of anti-VEGF-165b antibody,the expression of VEGF-165b protein was no significantly changed;and the down-regulation of the expression of ZO-1 was more obvious;EB leakage became more serious,peaking at 3d.EB analysis also showed that EB leakage in the peripheral retina was greater than that in the central retina.CONCLUSION:The endogenous VEGF-165b protein may protect the BRB from acute HIOP by regulating the expression of ZO-1.The differential destruction of BRB after acute HIOP may be related to the selective loss of retinal ganglion cells.
基金supported by the National Natural Science Foundation of China,Nos.82271419,81901902(to YZ),82202702(to ZW),82202351(to XH),82301550(to LYang),82271418(to XX)the Shanghai Rising-Star Program,No.22QA1408200(to YZ)the Fundamental Research Fundsfor the Central Universities,No.22120220555(to YZ).
文摘In the early stages of traumatic spinal cord injury,extensive accumulation of autophagosomes creates a neurotoxic microenvironment,exacerbating neuronal cell death and worsening tissue damage,ultimately hindering neurofunctional recovery.Activin A is a critical growth factor necessary for the development of the embryonic nervous system and for maintaining neuronal function in the adult cerebral cortex.It can inhibit excessive autophagy in ischemic stroke to reduce neuronal damage.However,the specific mechanism through which Activin A functions in the spinal cord remains poorly understood.In this study,we administered different concentrations of Activin A to neural stem cells from the spinal cord and found that Activin A stimulated the proliferation and neuronal differentiation of neural stem cells.Then,we established an in vitro oxidative stress model by using hydrogen peroxide to stimulate the neural stem cells-induced neurons.We found that Activin A could reduce apoptosis caused by oxidative stress.Subsequently,we treated a mouse model of spinal cord contusion with intrathecal injection of Activin A.Behavioral and electrophysiological results showed that Activin A promoted recovery of motor function and reconstruction of neural circuits in the model mice.Finally,RNA sequencing indicated that Activin A inhibited autophagy by activating the PI3K/AKT/mTOR pathway and upregulating the expression of synaptogenesis-related factor Sema3A in the spinal cord.These results suggest that Activin A may mediate the excessive autophagic response after spinal cord injury,promote the reconstruction of damaged neural circuits,and restore neurological function in the injured spinal cord.
基金supported by the National Natural Science Foundational of China,Nos.U24A20692(to CJZ),82371355(to CJZ),and 82101414(to MH)National NaturalScience Foundational of China for Excellent Young Scholars,No.82022019(to CJZ)+5 种基金Sichuan Special Fund for Distinguished Young Scholars,No.24NSFJQ0052(to CJZ)The Innovationand Entrepreneurial Team of Sichuan Tianfu Emei Program,No.CZ2024018(to CJZ)Funding for Distinguished Young Scholars of Sichuan Provincial People’sHospital,No.30420230005Funding for Distinguished Young Scholars of University of Electronic Science and Technology of China,No.A1098531023601381(toCJZ)Sichuan Science and Technology Support Project,No.2023YFS0212(to BH)Project of Sichuan Provincial Health Commission,No.19PJ265(to LD).
文摘Multiple sclerosis is a severe autoimmune disorder that is mainly mediated by pathogenic cluster of CD4^(+)T cell subsets.Despite advancements in the management of multiple sclerosis,there is a critical need for more effective and safer treatments.In the present study,we administered Lycium barbarum glycopeptide to a mouse model of experimental autoimmune encephalomyelitis-an animal model of multiple sclerosis-and evaluated its effects on pathogenic CD4^(+)T cell activation both in vivo and in vitro.Lycium barbarum glycopeptide significantly mitigated the clinical severity of experimental autoimmune encephalomyelitis,as demonstrated by reduced demyelination and neuroinflammation.Moreover,Lycium barbarum glycopeptide treatment decreased the infiltration of peripheral leukocytes into the central nervous system and suppressed pro-inflammatory cytokine expression.Lycium barbarum glycopeptide also modulated pathogenic CD4^(+)T cell activation by inhibiting T helper 1/T helper 17 cell differentiation while promoting regulatory T cell expansion.Notably,no side effects were observed,suggesting the long-term safety and tolerability of Lycium barbarum glycopeptide.Furthermore,RNA sequencing data indicated that Lycium barbarum glycopeptide inhibits activator protein-1,an essential regulator of T cell activation and differentiation.This finding was supported by the reversal of T helper/T helper 17 cell response suppression upon AP-1 blockade.Collectively,these results highlight the potential of Lycium barbarum glycopeptide as an innovative therapeutic agent for CD4^(+)T cell-associated autoimmune or inflammatory diseases,such as multiple sclerosis.
文摘目的研究VEGF165b和VEGF165在胃癌及癌旁正常组织中的表达及胃癌组织中VEGF165b m RNA/VEGF165 m RNA与预后的关系。方法分别应用实时RT-PCR和免疫组织化学法检测胃癌及癌旁正常组织中VEGF165b和VEGF165的基因及蛋白表达,并对患者进行2年的随访,比较VEGF165b m RNA/VEGF165 m RNA比值及这两种蛋白表达水平在胃癌及癌旁组织中的变化;同时分析VEGF165b与VEGF165蛋白表达的相关性以及胃癌组织中VEGF165b m RNA/VEGF165 m RNA与预后的关系。结果在胃癌组织中VEGF165b m RNA/VEGF165 m RNA低于癌旁正常组织中的水平;在癌旁正常组织中VEGF165b基因及蛋白表达强于VEGF165,在胃癌组织中VEGF165b基因及蛋白的表达则低于VEGF165,而且两者表达呈负相关关系;此外,在两年内死亡的患者中VEGF165b m RNA/VEGF165 m RNA明显低于生存患者中的比值水平。结论与正常组织相比,胃癌组织中存在VEGF165b向VEGF165基因及蛋白水平上的优势转换,这种变化有望成为治疗胃癌的新靶点。
文摘目的:对比观察VEGF165b重组蛋白(recombinant human VEGF165b protein,rhVEGF165b)与贝伐珠单抗对人胃癌裸鼠移植瘤生长的影响及作用机制.方法:用人胃癌细胞BGC-823接种于裸鼠皮下,建立裸鼠移植瘤模型,随机分为3组,每组21只:rhVEGF165b组(腹腔注射rhVEGF165b 10?g/kg)、贝伐珠单抗组(腹腔注射贝伐珠单抗5 mg/kg)、对照组(腹腔注射生理盐水10 mL/kg),分别于1、2、3 wk测量裸鼠移植瘤体积及瘤质量,免疫组织化学方法检测肿瘤组织中CD34表达(以阳性细胞数计算肿瘤微血管密度),TUNEL方法检测肿瘤组织细胞凋亡.结果:贝伐珠单抗组移植瘤体积(cm3)及瘤质量(g)在第1、2、3周均小于对照组(1 wk:0.453±0.119 vs 0.637±0.084,0.32±0.097vs 0.46±0.093;2 wk:1.691±0.381 vs 2.238±0.29,1.168±0.524 vs 2.013±0.833;3 wk:1.709±0.474 vs 4.872±0.594,1.747±0.557 vs3.463±0.986,均P<0.05),而rhVEGF165b组仅在第1、2周较对照组小(1 wk:0.546±0.132 vs0.637±0.084,1.894±0.599 vs 0.46±0.093;2wk:1.894±0.599 vs 2.238±0.29,1.537±0.568vs 2.013±0.833,均P<0.05),第3周其体积及瘤质量大于贝伐珠单抗组(3 wk:3.843±1.339 vs1.709±0.474,3.066±1.281 vs 1.747±0.557,P<0.05),与对照组无明显差异.rhVEGF165b组、贝伐珠单抗组CD34的表达水平在第1、2、3周均低于对照组(P<0.05,P<0.01),凋亡指数高于对照组(P<0.05,P<0.01);与贝伐珠单抗组相比,第1周rhVEGF165b组CD34的表达水平更低(P<0.05),但第2、3周却高于贝伐珠单抗组(P<0.01);rhVEGF165b组的凋亡指数在第1周明显高于贝伐珠单抗组(P<0.01),而第2、3周却低于贝伐珠单抗组(P<0.05).结论:rhVEGF165b、贝伐珠单抗对人胃癌移植瘤的生长有明显抑制作用,rhVEGF165b早期抑制血管生成、促进细胞凋亡表现更为明显.
文摘目的:初步探讨VEGF165b与膀胱移行细胞癌(Transitional cell carcinoma of the bladder,TCCB)发生发展的关系。方法:用S-P免疫组化法检测38例TCCB组织和36例正常膀胱组织中VEGF165b蛋白表达;以RT-PCR法检测55例TCCB组织和43例正常膀胱组织中VEGF165bmRNA表达。结果:正常膀胱组织中VEGF165b蛋白表达率为97.22%(35/36),TCCB组织中VEGF165b蛋白表达率为21.05%(8/38),表达差异有显著性(P<0.05);正常膀胱组织中VEGF165bmRNA阳性表达率为95.35%(41/43),明显高于TCCB组织中VEGF165bmRNA阳性表达率18.18%(10/55)(P<0.05);TCCB中VEGF165b蛋白和mRNA的表达与TCCB不同分期、分级间的表达差异具有统计学意义(P<0.05)。结论:VEGF165b在TCCB组织中的表达明显低于正常膀胱组织中的表达,这种表达差异可能与VEGF165b抑制膀胱癌的发生、发展作用有关。
