AIM: To facilitate close contacts between transplanted cardiomyocytes and host skeletal muscle using cell fusion mediated by hemagglutinating virus of Japan envelope(HVJ-E) and tissue maceration. METHODS: Cardiomyocyt...AIM: To facilitate close contacts between transplanted cardiomyocytes and host skeletal muscle using cell fusion mediated by hemagglutinating virus of Japan envelope(HVJ-E) and tissue maceration. METHODS: Cardiomyocytes(1.5 × 106) from fetal rats were first cultured. After proliferation, some cells were used for fusion with adult muscle fibers using HVJ-E. Other cells were used to create cardiomyocyte sheets(area: about 3.5 cm2 including 2.1 × 106 cells), which were then treated with Nile blue, separated, and transplanted between the latissimus dorsi and intercostal muscles of adult rats with four combinations of HVJ-E and/or Na OH maceration: G1: HVJ-E(+), Na OH(+), Cardiomyocytes(+); G2: HVJ-E(-), NaO H(+), Cardiomyocytes(+); G3: HVJ-E(+),Na OH(-), Cardiomyocytes(+); G4: HVJ-E(-), Na OH(-), Cardiomyocytes(-). At 1 and 2 wk after transplantation, the four groups were compared by detection of beating domains, motion images using moving target analysis software, action potentials, gene expression of MLC-2v and Mesp1 by reverse transcription-polymerase chain reaction, hematoxylin-eosin staining, and immunostaining for cardiac troponin and skeletal myosin.RESULTS: In vitro cardiomyocytes were fused with skeletal muscle fibers using HVJ-E. Cardiomyocyte sheets remained in the primary transplanted sites for 2 wk. Although beating domains were detected in G1, G2, and G3 rats, G1 rats prevailed in the number, size, motion image amplitudes, and action potential compared with G2 and G3 rats. Close contacts were only found in G1 rats. At 1 wk after transplantation, the cardiomyocyte sheets showed adhesion at various points to the myoblast layer in the latissimus dorsi muscle. At 2 wk after transplantation, close contacts were seen over a broad area. Part of the skeletal muscle sarcoplasma seemed to project into the myocardiocyte plasma and some nuclei appeared to share both sarcoplasmas.CONCLUSION: The present results show that close contacts were acquired and facilitated the beating function, thereby providing a new cel展开更多
The visualization and evaluation of fruit vascular bundles in grape are indispensable for understanding the development and growth of the fruit. The vascular bundles in grape are as thin as human hair, and strongly ad...The visualization and evaluation of fruit vascular bundles in grape are indispensable for understanding the development and growth of the fruit. The vascular bundles in grape are as thin as human hair, and strongly adhere to flesh cells, making it difficult to isolate individual intact vessel elements. Currently there is little information about methods to characterize grape vascular tissue. In this study, we describe an easy and effective technique to visualize the xylem cell structure of the ‘Red Globe' grape(Vitis L.). The intact vascular bundles of the grape were isolated through enzymatic degradation of flesh cells by hydrolases which were effective in removing flesh cells from vascular bundles. This enabled the illustration of the fine surface structure of vessel elements and their arrangement in the vascular bundles of the grape. This modified method to separate the vascular bundles into individual vessel elements was more effective than former methods of manually shaking to isolate individual vessels. Clear images of xylem vessel arrangement and structural characteristics of individual vessel element were acquired by light microscopy(BX51, Olympus, Tokyo, Japan), transmission electron microscopy(Tecnai 12, Philips, Netherlands) and scanning electron microscopy(GeminiSEM 300, Carl Zeiss, Germany). In addition, the 3D structure of vessel elements was observed using confocal laser scanning microscopy(Zeiss: model: LSM-880, Oberkochen, Germany). The imaging techniques for visualizing and analyzing the structure of xylem vessels in grape described in this study are an improvement of past methods. An effective method to isolate grape vascular bundles from flesh cells was also developed, which enables the imaging of the fine surface structure of vessel elements and their arrangement in grape vascular bundles. By adjusting the process of maceration of vascular bundles, an easy and effective method was developed to gently disrupt the plant tissue and isolate individual vessel elements. These improved techniques 展开更多
Background: Artemisia afra is an indigenous plant used in Africa. It is used in traditional medicine to treat a variety of diseases caused by bacterial infections. As a result, developing innovative therapeutic method...Background: Artemisia afra is an indigenous plant used in Africa. It is used in traditional medicine to treat a variety of diseases caused by bacterial infections. As a result, developing innovative therapeutic methods using natural ingredients to combat pathogenic germs is critical. Methodology: Artemisia afra leaves were extracted using maceration extraction utilizing three solvents (petroleum ether, ethanol, and dichloromethane). Fresh leaves were extracted using hydrodistillation. The agar well diffusion method was used to test the crude extract and essential oil leaves of Artemisia afra against different human pathogenic bacteria strains (E. coli, S. aureus, E. faecalis, and K. pneumonia) at different concentrations in the presence of a positive (ciprofloxacin 5.0 μg) and negative (DMSO) control. Results: Artemisia afra revealed the presence of alkaloids, flavonoids, tannins, saponins, anthraquinones, terpenoids, coumarins, phenolic quinones, cardiac glycosides, and steroids. It showed that the highest antibacterial activity given by the ethanol extract had highly inhibition zones against gram-positive and gram-negative bacteria. The essential oil extract was effective against all tested bacteria. Conclusion: Plant crude extracts and essential oils may have antibacterial properties due to the synergistic activity of two or more active secondary metabolites.展开更多
The effect of lanthanum on the growth and extracellular enzyme activity of Erwinia chrysanthemi(Ech) was studied. Lanthanum inhibited the growth of Ech in solid culture at the concentration of 50, 100, 150, 200, 250, ...The effect of lanthanum on the growth and extracellular enzyme activity of Erwinia chrysanthemi(Ech) was studied. Lanthanum inhibited the growth of Ech in solid culture at the concentration of 50, 100, 150, 200, 250, 300 and 350 mgL-1, and the inhibition increased with the increase of the RE concentration. The formation of colony delayed and the diameter of colony decreased. Lanthanum stimulated the growth of Ech within 24h when the lanthanum concentration was less than 200 mgL-1 in liquid medium, but significantly inhibited the growth of Ech with the increase of lanthanum concentration and culture time. When the concentration was higher than 350 mgL-1, the growth of Ech was inhibited completely both in solid and liquid culture. The extracellular enzyme acitivity of Ech was affected by lanthanum at 200 mgL-1. The activity of cellulose was promoted more than that of protease but the activity of pectinase was decreased. The maceration to potato tuber tissue decreased when the cell free filtrate was tr展开更多
基金Supported by A Grant-in-Aid for Scientific Research from the Japanese Ministry of Education,Science and Sports,No.24240076
文摘AIM: To facilitate close contacts between transplanted cardiomyocytes and host skeletal muscle using cell fusion mediated by hemagglutinating virus of Japan envelope(HVJ-E) and tissue maceration. METHODS: Cardiomyocytes(1.5 × 106) from fetal rats were first cultured. After proliferation, some cells were used for fusion with adult muscle fibers using HVJ-E. Other cells were used to create cardiomyocyte sheets(area: about 3.5 cm2 including 2.1 × 106 cells), which were then treated with Nile blue, separated, and transplanted between the latissimus dorsi and intercostal muscles of adult rats with four combinations of HVJ-E and/or Na OH maceration: G1: HVJ-E(+), Na OH(+), Cardiomyocytes(+); G2: HVJ-E(-), NaO H(+), Cardiomyocytes(+); G3: HVJ-E(+),Na OH(-), Cardiomyocytes(+); G4: HVJ-E(-), Na OH(-), Cardiomyocytes(-). At 1 and 2 wk after transplantation, the four groups were compared by detection of beating domains, motion images using moving target analysis software, action potentials, gene expression of MLC-2v and Mesp1 by reverse transcription-polymerase chain reaction, hematoxylin-eosin staining, and immunostaining for cardiac troponin and skeletal myosin.RESULTS: In vitro cardiomyocytes were fused with skeletal muscle fibers using HVJ-E. Cardiomyocyte sheets remained in the primary transplanted sites for 2 wk. Although beating domains were detected in G1, G2, and G3 rats, G1 rats prevailed in the number, size, motion image amplitudes, and action potential compared with G2 and G3 rats. Close contacts were only found in G1 rats. At 1 wk after transplantation, the cardiomyocyte sheets showed adhesion at various points to the myoblast layer in the latissimus dorsi muscle. At 2 wk after transplantation, close contacts were seen over a broad area. Part of the skeletal muscle sarcoplasma seemed to project into the myocardiocyte plasma and some nuclei appeared to share both sarcoplasmas.CONCLUSION: The present results show that close contacts were acquired and facilitated the beating function, thereby providing a new cel
基金supported by the National Natural Science Foundation of China (Grant Nos. 31872050 and 32102348)。
文摘The visualization and evaluation of fruit vascular bundles in grape are indispensable for understanding the development and growth of the fruit. The vascular bundles in grape are as thin as human hair, and strongly adhere to flesh cells, making it difficult to isolate individual intact vessel elements. Currently there is little information about methods to characterize grape vascular tissue. In this study, we describe an easy and effective technique to visualize the xylem cell structure of the ‘Red Globe' grape(Vitis L.). The intact vascular bundles of the grape were isolated through enzymatic degradation of flesh cells by hydrolases which were effective in removing flesh cells from vascular bundles. This enabled the illustration of the fine surface structure of vessel elements and their arrangement in the vascular bundles of the grape. This modified method to separate the vascular bundles into individual vessel elements was more effective than former methods of manually shaking to isolate individual vessels. Clear images of xylem vessel arrangement and structural characteristics of individual vessel element were acquired by light microscopy(BX51, Olympus, Tokyo, Japan), transmission electron microscopy(Tecnai 12, Philips, Netherlands) and scanning electron microscopy(GeminiSEM 300, Carl Zeiss, Germany). In addition, the 3D structure of vessel elements was observed using confocal laser scanning microscopy(Zeiss: model: LSM-880, Oberkochen, Germany). The imaging techniques for visualizing and analyzing the structure of xylem vessels in grape described in this study are an improvement of past methods. An effective method to isolate grape vascular bundles from flesh cells was also developed, which enables the imaging of the fine surface structure of vessel elements and their arrangement in grape vascular bundles. By adjusting the process of maceration of vascular bundles, an easy and effective method was developed to gently disrupt the plant tissue and isolate individual vessel elements. These improved techniques
文摘Background: Artemisia afra is an indigenous plant used in Africa. It is used in traditional medicine to treat a variety of diseases caused by bacterial infections. As a result, developing innovative therapeutic methods using natural ingredients to combat pathogenic germs is critical. Methodology: Artemisia afra leaves were extracted using maceration extraction utilizing three solvents (petroleum ether, ethanol, and dichloromethane). Fresh leaves were extracted using hydrodistillation. The agar well diffusion method was used to test the crude extract and essential oil leaves of Artemisia afra against different human pathogenic bacteria strains (E. coli, S. aureus, E. faecalis, and K. pneumonia) at different concentrations in the presence of a positive (ciprofloxacin 5.0 μg) and negative (DMSO) control. Results: Artemisia afra revealed the presence of alkaloids, flavonoids, tannins, saponins, anthraquinones, terpenoids, coumarins, phenolic quinones, cardiac glycosides, and steroids. It showed that the highest antibacterial activity given by the ethanol extract had highly inhibition zones against gram-positive and gram-negative bacteria. The essential oil extract was effective against all tested bacteria. Conclusion: Plant crude extracts and essential oils may have antibacterial properties due to the synergistic activity of two or more active secondary metabolites.
文摘The effect of lanthanum on the growth and extracellular enzyme activity of Erwinia chrysanthemi(Ech) was studied. Lanthanum inhibited the growth of Ech in solid culture at the concentration of 50, 100, 150, 200, 250, 300 and 350 mgL-1, and the inhibition increased with the increase of the RE concentration. The formation of colony delayed and the diameter of colony decreased. Lanthanum stimulated the growth of Ech within 24h when the lanthanum concentration was less than 200 mgL-1 in liquid medium, but significantly inhibited the growth of Ech with the increase of lanthanum concentration and culture time. When the concentration was higher than 350 mgL-1, the growth of Ech was inhibited completely both in solid and liquid culture. The extracellular enzyme acitivity of Ech was affected by lanthanum at 200 mgL-1. The activity of cellulose was promoted more than that of protease but the activity of pectinase was decreased. The maceration to potato tuber tissue decreased when the cell free filtrate was tr
