In situ direct reprogramming technology can directly convert endogenous glial cells into functional neurons in vivo for central nervous system repair. Polypyrimidine tract-binding protein 1(PTB) knockdown has been sho...In situ direct reprogramming technology can directly convert endogenous glial cells into functional neurons in vivo for central nervous system repair. Polypyrimidine tract-binding protein 1(PTB) knockdown has been shown to reprogram astrocytes to functional neurons in situ. In this study, we used AAV-PHP.e B-GFAP-sh PTB to knockdown PTB in a mouse model of ischemic stroke induced by endothelin-1, and investigated the effects of GFAP-sh PTB-mediated direct reprogramming to neurons. Our results showed that in the mouse model of ischemic stroke, PTB knockdown effectively reprogrammed GFAP-positive cells to neurons in ischemic foci, restored neural tissue structure, reduced inflammatory response, and improved behavioral function. These findings validate the effectiveness of in situ transdifferentiation of astrocytes, and suggest that the approach may be a promising strategy for stroke treatment.展开更多
AIM： To study the effect of senescence marker protein 30（SMP30） on the proliferation and apoptosis of human lens epithelial cell（HLEC） SRA01/04.METHODS： SMP30 overexpression（OE） and knock down（KD） type cell ...AIM： To study the effect of senescence marker protein 30（SMP30） on the proliferation and apoptosis of human lens epithelial cell（HLEC） SRA01/04.METHODS： SMP30 overexpression（OE） and knock down（KD） type cell lines were cultivated by using two groups regucalcin（RGN; SMP30） lentiviral vectors（LVRGN, LV-RGN-RNAi） and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction（q-PCR） analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8（CCK8） assay to measure cell viability and 5-bromodeoxyuridine（Brd U） assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04.RESULTS： We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation（P〈0.05） compared with the control group, and the KD group inhibited cell proliferation（P〈0.05）. The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group（P〈0.05） but lower in OE group（P〈0.01）. The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group. CONCLUSION： Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.展开更多
The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated...The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated through Genefishing analysis during defense responses to Ralstonia solanacearum.The cDNA of RPS 30 contained a 189 base pair(bp)open-reading frame encoding 62 amino acids.The genomic DNA consists of 272 bp containing two exons and one 83 bp intron.The RPS 30 mRNA transcript was mainly expressed in roots and leaves.The expression level of the RPS 30 mRNA transcripts was up-regulated sharply 6 h after bacterial challenge and was 12 times greater than that of the control group.The phylogenetic analysis for genes encoding proteins showed that RPS30 were conserved within dicotyledonous and monocotyledonous plants.d S extremely exceeded d N in all branches of the tree(d N/d S<1.0),indicating that functional constraint have acted on RPS 30 throughout evolution.展开更多
4PU—30[N—phenyl—’N—(2—chloro—4—pyridyl) urea] is a new type of plant growth regulator with cytokinin properties. It has been confirmed to delay rice leaf senescence effectively. In order to elucidate the physi...4PU—30[N—phenyl—’N—(2—chloro—4—pyridyl) urea] is a new type of plant growth regulator with cytokinin properties. It has been confirmed to delay rice leaf senescence effectively. In order to elucidate the physiological role of 4PU—30 in delaying senescence, the changes of protein, nucleic acid contents, and the related activities of degradative enzymes were studied. Shanyou 63, an indica hybrid rice was used for this experiment. In the in vitro experiment, two full—developed leaves from the top during heading stage were collected and cut into 5.0cm segments, They were floated on the surface of distilled water containing 0.1mg/14PU—30 and incubated in darkness at 30 C. The leaves floated on distilled water were used as control.It was observed that chlorophyll content in controlled leaves declined rapidly started from the second day and dropped by 93.4% on the 6th day while that in leaves treated with 4PU—30 declined by 41.4% only. During senescence, specific activities of hemoglobin—digesting展开更多
目的:研究Tat作用蛋白30(Tat-interactive protein 30,TIP30)基因在胃癌组织中的表达及其对胃癌血管生成的影响。方法:运用免疫组织化学方法检测52例胃癌组织及47例癌旁正常组织中TIP30蛋白的表达水平,同时检测CD34标记的微血管密度(mic...目的:研究Tat作用蛋白30(Tat-interactive protein 30,TIP30)基因在胃癌组织中的表达及其对胃癌血管生成的影响。方法:运用免疫组织化学方法检测52例胃癌组织及47例癌旁正常组织中TIP30蛋白的表达水平,同时检测CD34标记的微血管密度(microvessel density,MVD)。结果:TIP30在胃癌和癌旁正常组织中的阳性率分别为53.8%和85.1%,两者相比较差异有统计学意义(χ2=11.22,P﹤0.01),TIP30表达水平与患者年龄、性别、病理分化程度及肿瘤大小无关,与有无淋巴结转移及TNM分期有关。胃癌组织和癌旁正常组织中MVD值分别为27.37±2.68和21.87±4.11,两者比较差异有统计学意义(t=7.95,P<0.01),胃癌组织中TIP30表达阳性者较阴性者MVD值显著降低(t=-3.18,P=0.003)。结论:TIP30在胃癌组织中表达降低,其可能通过抑制胃癌血管生成影响胃癌的发生发展。展开更多
目的观察高表达的衰老标记蛋白30(SMP30)对过氧化氢(H_2O_2)所致的正常人皮肤成纤维细胞(NHSF)衰老表型的影响。方法实验分为转染H_2O_2处理组(NHSF+SMP30+H_2O_2组)、转空载体H_2O_2处理组(NHSF+空载体+H_2O_2组)和转空载体组(NHSF+空...目的观察高表达的衰老标记蛋白30(SMP30)对过氧化氢(H_2O_2)所致的正常人皮肤成纤维细胞(NHSF)衰老表型的影响。方法实验分为转染H_2O_2处理组(NHSF+SMP30+H_2O_2组)、转空载体H_2O_2处理组(NHSF+空载体+H_2O_2组)和转空载体组(NHSF+空载体组)。NHSF细胞分别转染人SMP30 c DNA或空载体pc DNA3.1(分别命名为NHSF+SMP30、NHSF+空载体),然后150μmol·L-1H_2O_2处理2 h。应用反转录-聚合酶链反应(RT-PCR)和蛋白印迹法分析SMP30的表达,显微镜观察细胞形态,并检测与衰老相关的β-半乳糖苷酶(SA-β-gal)、细胞活性氧(ROS)及超氧化物歧化酶(SOD)的活性。结果人SMP30 c DNA明显上调了NHSF中SMP30的表达。NHSF+空载体+H_2O_2组SMP30 mRNA和蛋白表达水平明显低于NHSF+空载体组(P<0.05);NHSF+SMP30+H_2O_2组SMP30 mRNA和蛋白表达水平高于NHSF+空载体+H_2O_2组(P<0.05)。NHSF+空载体+H_2O_2组较NHSF+空载体组SA-β-gal染色率明显增高(P<0.05);NHSF+SMP30+H_2O_2组较NHSF+空载体+H_2O_2组SA-β-gal染色率明显下降(P<0.05)。NHSF+空载体+H_2O_2组较NHSF+空载体组ROS活性明显增高(P<0.05),而SOD水平明显降低(P<0.05);NHSF+SMP30+H_2O_2组较NHSF+空载体+H_2O_2组ROS活性明显下降,而SOD水平明显上升(P<0.05)。结论高表达的SMP30能够抑制细胞SA-β-gal和ROS的表达,同时上调SOD的活性。展开更多
基金supported by the National Natural Science Foundation of China,No.82071418the Natural Science Foundation of Fujian Province,No.2020J01612 (both to EH)。
文摘In situ direct reprogramming technology can directly convert endogenous glial cells into functional neurons in vivo for central nervous system repair. Polypyrimidine tract-binding protein 1(PTB) knockdown has been shown to reprogram astrocytes to functional neurons in situ. In this study, we used AAV-PHP.e B-GFAP-sh PTB to knockdown PTB in a mouse model of ischemic stroke induced by endothelin-1, and investigated the effects of GFAP-sh PTB-mediated direct reprogramming to neurons. Our results showed that in the mouse model of ischemic stroke, PTB knockdown effectively reprogrammed GFAP-positive cells to neurons in ischemic foci, restored neural tissue structure, reduced inflammatory response, and improved behavioral function. These findings validate the effectiveness of in situ transdifferentiation of astrocytes, and suggest that the approach may be a promising strategy for stroke treatment.
基金Supported by the National Natural Science Foundation of China(No.81360146)
文摘AIM： To study the effect of senescence marker protein 30（SMP30） on the proliferation and apoptosis of human lens epithelial cell（HLEC） SRA01/04.METHODS： SMP30 overexpression（OE） and knock down（KD） type cell lines were cultivated by using two groups regucalcin（RGN; SMP30） lentiviral vectors（LVRGN, LV-RGN-RNAi） and the respective negative control virus infect SRA01/04 cells. Western blot and real-time quantitative polymerase chain reaction（q-PCR） analysis were used to determine RGN overexpression and knock down efficiency. We use cell counting kit-8（CCK8） assay to measure cell viability and 5-bromodeoxyuridine（Brd U） assay to test cell proliferation. Cell cycle was measured by PI FACS assay and cell apoptosis was tested by Annexin V-APC assay through flow cytometry. We use Western blot to measure the content of caspase-3 in SRA01/04.RESULTS： We used PCR and Western blot techniques to determine the successful transfection of SMP30 OE and KD SRA01/04 cell lines. By CCK8, Brdu and PI FACS cell cycle assay, it was found that the SMP30 OE group promoted cell proliferation（P〈0.05） compared with the control group, and the KD group inhibited cell proliferation（P〈0.05）. The results of Annexin V-APC signal staining detection indicated that compared with respective control group, the cell apoptosis rate was higher in KD group（P〈0.05） but lower in OE group（P〈0.01）. The expression of caspase-3 was down-regulated in OE group through Western blot assay and up-regulated in KD group compared with respective control group. CONCLUSION： Proliferation of SRA01/04 was promoted by SMP30 OE and apoptosis was suppressed. Increasing the expression of SMP30 may protect HLEC SRA01/04 against apoptosis in cataract.
基金Natural Science Funds for Young Scholar of Shandong Academy of Agricultural Sciences,China(No.2015YQN13)Natural Science Foundation of Shandong Province,China(ZR2015YL064)+2 种基金Qingdao Science and Technology Plan Basic Research Project,China(No.12-1-4-11-(1)-jch)China Agricultural Research System(No.CARS-13)Agricultural Scientific and Technological Innovation Project of Shandong Academy of Agricultural Sciences,China(No.CXGC2018E21)
文摘The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated through Genefishing analysis during defense responses to Ralstonia solanacearum.The cDNA of RPS 30 contained a 189 base pair(bp)open-reading frame encoding 62 amino acids.The genomic DNA consists of 272 bp containing two exons and one 83 bp intron.The RPS 30 mRNA transcript was mainly expressed in roots and leaves.The expression level of the RPS 30 mRNA transcripts was up-regulated sharply 6 h after bacterial challenge and was 12 times greater than that of the control group.The phylogenetic analysis for genes encoding proteins showed that RPS30 were conserved within dicotyledonous and monocotyledonous plants.d S extremely exceeded d N in all branches of the tree(d N/d S<1.0),indicating that functional constraint have acted on RPS 30 throughout evolution.
文摘4PU—30[N—phenyl—’N—(2—chloro—4—pyridyl) urea] is a new type of plant growth regulator with cytokinin properties. It has been confirmed to delay rice leaf senescence effectively. In order to elucidate the physiological role of 4PU—30 in delaying senescence, the changes of protein, nucleic acid contents, and the related activities of degradative enzymes were studied. Shanyou 63, an indica hybrid rice was used for this experiment. In the in vitro experiment, two full—developed leaves from the top during heading stage were collected and cut into 5.0cm segments, They were floated on the surface of distilled water containing 0.1mg/14PU—30 and incubated in darkness at 30 C. The leaves floated on distilled water were used as control.It was observed that chlorophyll content in controlled leaves declined rapidly started from the second day and dropped by 93.4% on the 6th day while that in leaves treated with 4PU—30 declined by 41.4% only. During senescence, specific activities of hemoglobin—digesting
文摘目的:研究Tat作用蛋白30(Tat-interactive protein 30,TIP30)基因在胃癌组织中的表达及其对胃癌血管生成的影响。方法:运用免疫组织化学方法检测52例胃癌组织及47例癌旁正常组织中TIP30蛋白的表达水平,同时检测CD34标记的微血管密度(microvessel density,MVD)。结果:TIP30在胃癌和癌旁正常组织中的阳性率分别为53.8%和85.1%,两者相比较差异有统计学意义(χ2=11.22,P﹤0.01),TIP30表达水平与患者年龄、性别、病理分化程度及肿瘤大小无关,与有无淋巴结转移及TNM分期有关。胃癌组织和癌旁正常组织中MVD值分别为27.37±2.68和21.87±4.11,两者比较差异有统计学意义(t=7.95,P<0.01),胃癌组织中TIP30表达阳性者较阴性者MVD值显著降低(t=-3.18,P=0.003)。结论:TIP30在胃癌组织中表达降低,其可能通过抑制胃癌血管生成影响胃癌的发生发展。
文摘目的观察高表达的衰老标记蛋白30(SMP30)对过氧化氢(H_2O_2)所致的正常人皮肤成纤维细胞(NHSF)衰老表型的影响。方法实验分为转染H_2O_2处理组(NHSF+SMP30+H_2O_2组)、转空载体H_2O_2处理组(NHSF+空载体+H_2O_2组)和转空载体组(NHSF+空载体组)。NHSF细胞分别转染人SMP30 c DNA或空载体pc DNA3.1(分别命名为NHSF+SMP30、NHSF+空载体),然后150μmol·L-1H_2O_2处理2 h。应用反转录-聚合酶链反应(RT-PCR)和蛋白印迹法分析SMP30的表达,显微镜观察细胞形态,并检测与衰老相关的β-半乳糖苷酶(SA-β-gal)、细胞活性氧(ROS)及超氧化物歧化酶(SOD)的活性。结果人SMP30 c DNA明显上调了NHSF中SMP30的表达。NHSF+空载体+H_2O_2组SMP30 mRNA和蛋白表达水平明显低于NHSF+空载体组(P<0.05);NHSF+SMP30+H_2O_2组SMP30 mRNA和蛋白表达水平高于NHSF+空载体+H_2O_2组(P<0.05)。NHSF+空载体+H_2O_2组较NHSF+空载体组SA-β-gal染色率明显增高(P<0.05);NHSF+SMP30+H_2O_2组较NHSF+空载体+H_2O_2组SA-β-gal染色率明显下降(P<0.05)。NHSF+空载体+H_2O_2组较NHSF+空载体组ROS活性明显增高(P<0.05),而SOD水平明显降低(P<0.05);NHSF+SMP30+H_2O_2组较NHSF+空载体+H_2O_2组ROS活性明显下降,而SOD水平明显上升(P<0.05)。结论高表达的SMP30能够抑制细胞SA-β-gal和ROS的表达,同时上调SOD的活性。
