AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelia...AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity.METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both were added to the system. The wells (n = 8/group) were digitally photographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated.RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina. The angiogenic effects of bFGF or VEGF were dose-clependent in bhe range from 10 to 40 ng/mL. At d 1, 10 ng/mL of bFGF and VEGF induced angiogenesis with an ALS of 32.13±16.6 μm and 43.75±27.92 μm, respectively, which were significantly higher than that of the control (5.88±4.45 μm, P<0.01),and the differences became more significant as the time increased. In addition, the combination of 10 ng/mL of bFGF and VEGF each induced a more significant effect than the summed effects of bFGF (10 ng/mL) alone and VEGF (10 ng/mL) alone when analyzed using SPSS system for general linear model (GLM) (P= 0.011), and bhat also exceeded the effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION: A microcarrier-based in vitro threedimensional angiogenesis model can be developed in fibrin.It offers a unique system for quantitative analysis of angiogenesis. Both bFGF and VEGF exert their angiogenic effects on HMVECs synergistically and in a dose-dependent manner.展开更多
AIM:To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogeness system with affymetrix oligonucleotide array. METHOD...AIM:To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogeness system with affymetrix oligonucleotide array. METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed. RESULTS: A total of 1 961 genes were up- or downreg-ulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-α, CXCL2/GRO-β, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, OOCL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis. CONCLUSION: The present study demonstrates a global profile of gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be conducted.展开更多
AIM Terminalia Catappa L. is a combretaceous plant broadly distributed on tropical and subtropical areas. The leaves of this plant have been used as a folk medicine for treating hepatitis in Indian, the Philippines et...AIM Terminalia Catappa L. is a combretaceous plant broadly distributed on tropical and subtropical areas. The leaves of this plant have been used as a folk medicine for treating hepatitis in Indian, the Philippines et al. Our previous studies showed that the extract of Terminalia Catappa L. leaves (TCCE) exerted antioxidative, anti - inflammatory and hepatoprotective activities, but the active substitutes and mechanisms remain unknown. The present research focused on investigating the antihepatotoxic activity and the new mitochondrial mechanisms of asiatic acid (AA), an important active component of TCCE,on acute liver injury. METHODS LPS^+D-GaIN induced-hepatic injury models was adopted to study the antihepatotoxic role of AA. The activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected, and the morphological change was observed. At the same time, the level of mRNA and protein of voltage dependent anion channel (VDAC), the most important componentprotein of mitochondrial permeability transition pore (PTP) in insulted liver was analyzed using RT-PCR and Western blot. Furthermore,展开更多
NF-κB is a transcription factor of eukaryote,whose family comprises five members in mammals and three in drosophila.Transcription factors of the NF-κB family are activated in response to signals that lead to cell gr...NF-κB is a transcription factor of eukaryote,whose family comprises five members in mammals and three in drosophila.Transcription factors of the NF-κB family are activated in response to signals that lead to cell growth,differentiation,apoptosis and other events.NF-κB takes part in expression of numerous cytokines and adhesion molecules which are critical elements involved in the regulation of immune responses.In this review, we focus on our current understanding of NF-κB signal pathway and its role in the innate and adaptive immune responses in which these transcription factors have a key regulatory function.Furthermore we review what is currently known about their effects associated with apoptosis.Cellular & Molecular Immunology.2004; 1(5):343-350.展开更多
Manipulation of mouse genome has merged as one of the most important approaches for studying genefunction and establishing the disease model because of the high homology between human genome and mouse genome. In this ...Manipulation of mouse genome has merged as one of the most important approaches for studying genefunction and establishing the disease model because of the high homology between human genome and mouse genome. In this study, the chemical mutagen ethylnitrosourea (ENU) was employed for inducing germ cell mutations in maleC57BL/6J mice. The first generation (G1) of the backcrossof these mutated mice, totally 3172, was screened for abnor-mal phenotypes on gross morphology, behavior, learning and memory, auditory brainstem response (ABR), electrocardio-gram (ECG), electroretinogram (ERG), flash-visual evoked potential (F-VEP), bone mineral density, and blood sugarlevel. 595 mice have been identified with specific dominantabnormalities. Fur color changes, eye defects and hearing loss occurred at the highest frequency. Abnormalities related to metabolism alteration are least frequent. Interestingly, eye defects displayed significant left-right asymmetry and sexpreference. Sex preference is also observed in mice with ab-normal bone mineral density. Among 104 G1 generation mutant mice examined for inheritability, 14 of them have been confirmed for passing abnormal phenotypes to their progenies. However, we did not observe behavior abnormali-ties of G1 mice to be inheritable, suggesting multi-gene con-trol for these complicated functions in mice. In conclusion, the generation of these mutants paves the way for under-standing molecular and cellular mechanisms of these ab-normal phenotypes, and accelerates the cloning of disease-related genes.展开更多
文摘AIM: To develop an in vitro three-dimensional (3-D) angiogenesis system to analyse the capillary sprouts induced in response to the concentration ranges of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) and to quantify their synergistic activity.METHODS: Microcarriers (MCs) coated with human microvascular endothelial cells (HMVECs) were embedded in fibrin gel and cultured in 24-well plates with assay media. The growth factors bFGF, or VEGF, or both were added to the system. The wells (n = 8/group) were digitally photographed and the average length of capillary-like sprouts (ALS) from each microcarrier was quantitated.RESULTS: In aprotinin-stabilized fibrin matrix, human microvascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina. The angiogenic effects of bFGF or VEGF were dose-clependent in bhe range from 10 to 40 ng/mL. At d 1, 10 ng/mL of bFGF and VEGF induced angiogenesis with an ALS of 32.13±16.6 μm and 43.75±27.92 μm, respectively, which were significantly higher than that of the control (5.88±4.45 μm, P<0.01),and the differences became more significant as the time increased. In addition, the combination of 10 ng/mL of bFGF and VEGF each induced a more significant effect than the summed effects of bFGF (10 ng/mL) alone and VEGF (10 ng/mL) alone when analyzed using SPSS system for general linear model (GLM) (P= 0.011), and bhat also exceeded the effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION: A microcarrier-based in vitro threedimensional angiogenesis model can be developed in fibrin.It offers a unique system for quantitative analysis of angiogenesis. Both bFGF and VEGF exert their angiogenic effects on HMVECs synergistically and in a dose-dependent manner.
文摘AIM:To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogeness system with affymetrix oligonucleotide array. METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed. RESULTS: A total of 1 961 genes were up- or downreg-ulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-α, CXCL2/GRO-β, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, OOCL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis. CONCLUSION: The present study demonstrates a global profile of gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be conducted.
文摘AIM Terminalia Catappa L. is a combretaceous plant broadly distributed on tropical and subtropical areas. The leaves of this plant have been used as a folk medicine for treating hepatitis in Indian, the Philippines et al. Our previous studies showed that the extract of Terminalia Catappa L. leaves (TCCE) exerted antioxidative, anti - inflammatory and hepatoprotective activities, but the active substitutes and mechanisms remain unknown. The present research focused on investigating the antihepatotoxic activity and the new mitochondrial mechanisms of asiatic acid (AA), an important active component of TCCE,on acute liver injury. METHODS LPS^+D-GaIN induced-hepatic injury models was adopted to study the antihepatotoxic role of AA. The activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were detected, and the morphological change was observed. At the same time, the level of mRNA and protein of voltage dependent anion channel (VDAC), the most important componentprotein of mitochondrial permeability transition pore (PTP) in insulted liver was analyzed using RT-PCR and Western blot. Furthermore,
文摘NF-κB is a transcription factor of eukaryote,whose family comprises five members in mammals and three in drosophila.Transcription factors of the NF-κB family are activated in response to signals that lead to cell growth,differentiation,apoptosis and other events.NF-κB takes part in expression of numerous cytokines and adhesion molecules which are critical elements involved in the regulation of immune responses.In this review, we focus on our current understanding of NF-κB signal pathway and its role in the innate and adaptive immune responses in which these transcription factors have a key regulatory function.Furthermore we review what is currently known about their effects associated with apoptosis.Cellular & Molecular Immunology.2004; 1(5):343-350.
基金supported by the State“863”High-Tech Project and the National Gongguan Project
文摘Manipulation of mouse genome has merged as one of the most important approaches for studying genefunction and establishing the disease model because of the high homology between human genome and mouse genome. In this study, the chemical mutagen ethylnitrosourea (ENU) was employed for inducing germ cell mutations in maleC57BL/6J mice. The first generation (G1) of the backcrossof these mutated mice, totally 3172, was screened for abnor-mal phenotypes on gross morphology, behavior, learning and memory, auditory brainstem response (ABR), electrocardio-gram (ECG), electroretinogram (ERG), flash-visual evoked potential (F-VEP), bone mineral density, and blood sugarlevel. 595 mice have been identified with specific dominantabnormalities. Fur color changes, eye defects and hearing loss occurred at the highest frequency. Abnormalities related to metabolism alteration are least frequent. Interestingly, eye defects displayed significant left-right asymmetry and sexpreference. Sex preference is also observed in mice with ab-normal bone mineral density. Among 104 G1 generation mutant mice examined for inheritability, 14 of them have been confirmed for passing abnormal phenotypes to their progenies. However, we did not observe behavior abnormali-ties of G1 mice to be inheritable, suggesting multi-gene con-trol for these complicated functions in mice. In conclusion, the generation of these mutants paves the way for under-standing molecular and cellular mechanisms of these ab-normal phenotypes, and accelerates the cloning of disease-related genes.
