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Objective: To study the effect of apigenin on the expression of microglia in penumbra and cerebral water content after acute transient focal cerebral ischemia-reperfution injury in rats. Methods: The acute transient focal cerebral ischemia-reperfusion models in rats were established with the modified suture method. Male Sprague-Dawley rats were randomly divided into Sham group, model group, and apigenin groups (there were 6, 24, 48, 72 h, and 7 d reperfusion treatments in the model and apigenin groups, n = 12). All of them are 11 groups. The neurological behavior scores were valued. By FITC labeled isolectin B4 (FITC-ILB4) histochemistry staining, the infiltration of monocytes and the changes of cell morphology and number of brain-derived microglia in penumbra of six rats in every group were observed under laser scanning confocal microscope (LSCM). Water content was measured in isolated brain tissue of other six rats. Results: The positive cells of ILB4 (ILB4+) including microglia cells (Rhod 6G-) and infiltration of monocytes (Rhod 6G+) were found in cerebral ischemic area around penumbra of rats after 6 h ischemia-reperfusion in model group; The morphology changed to Amoeba-like; Microglia increased significantly after 48 h and reached to peak in 72 h, which mainly belonged to the proliferation of brain-derived microglia in Amoeba-like morphology. Microglia cell decreased in 7 d, and microglia in apigenin group obviously decreased more than that in model group (P<.05, 0.01) with the similar morphological change in corresponding time points. In 48 and 72 h of cerebral ischimia, the water content in brain tissue of rats in apigenin group was markedly lower than that in model (P<0.01). There was negative line correlation between the neurological behavior score and the number of ILB4 + cells in penumbra of model group (r=-0.415, P<.05). Apigenin could reduce the degree of neurological deficiency in model group and mitigate the brain injury effectively. Conclusion: A part of microglia cells inpenumbra are associated with brain injury; Apigenin shows the protection on acute transient focal cerebral ischemia-reperfution injury in rats, which maybe relates with down-regulating the microglia cell number and inhibiting the excessive activation of microglia cell.
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Objective: To observe the biocompatibility of titanium implant surfaces using laser scanning confocal microscopy (LSCM). Methods; Titanium implant surfaces were prepared by 3 different methods. Osteoblasts were seeded onto the titanium surfaces. The cells were investigated by LSCM. Results: The sand-blasted titanium surfaces showed the best biocompatibility. Conclusion; The biocompatibility of titanium implant surfaces depends on the surface preparation method.
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Objective To explore the double-labeling method of monitoring the GHRP regulatory function on [Ca2+]i and NO in cardiomyocytes of rats on real time under LSCM.Methods The reformed constant-flow Langendorff system and enzyme-dissociated was used to isolate cardiomyocytes.[Ca2+]i and NO in the cardiomyocytes of SD rats were double-labeled by their molecular probe Rhod-2/AM and DAF-FM/DA,respectively to monitor the regulatory function of GHRP on [Ca2+]i and NO on real time by LSCM.Results Ca2+ signal showed a red fluorescence and NO showed a green fluorescence while the overlapping of the two signals showed a yellow-green fluorescence by this system,and the similar effect presents in both double-labeled state and the single labeled one:GHRP induced a transient[Ca2+]i increase then followed by a plateau phase while there was not significant change in NO signal system after GHRP stimulation under the LSCM in the cardiomyocytes of rats.Conclusions After having established the double-labeling method we monitored the GHRP regulatory function on [Ca2+]i and NO on real time in cardiomyocytes of rats under LSCM causing the [Ca2+]i biphasic increase while no significant change in NO signal system.
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Objective To find out the method of ET-1 in regulating trabecular meshwork cell(TMC) contraction and to explore new effective drug treating glaucoma with little side-effects.Methods TMCs were obtained through the cultured tissue,and then loaded with Fluo-3/AM,the value of [Ca~(2+)]i was obtained by dynamically scanning the changes of intracellular fluorescent intensity after the application of different concentration of ET-1.Results The application of ET-1(10~(-9),10~(-8)and 10~(-7)mol/L),led to an increase in([Ca~(2+)]i.) Especially,the application of 10~(-8) mol/L ET-1 led to a significant increase in [Ca~(2+)]i: from(327.50)?(24.57) to 373.60?40.18(n=8,P
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Objective To observe effects of Glycyrrhiza flavonoids on DNA and RNA in tumor cell of S_(180) and H_(22) tumor-bearing mice.Methods S_(180) and H_(22) mice were randomly divided into Glycyrrhiza flavonoids(25,11.25,and 5.58 mg/kg) groups,positive control(cytoxan 25 mg/kg) group,and negative control(NS) group,whom were given drugs by sc.DNA and RNA in tumor cells were examined,respectively by Laser Scanning Confocal Microscope and fluorescent probe of acridine orange(AO) technology.Results All different dosage of Glycyrrhiza flavonoids,cytoxan reduced the brightness of fluorescence of DNA and RNA;low and high dosage of Glycyrrhiza flavonoids had no significant effect on the fluorescence pixels of RNA/DNA;Both middle dosage of Glycyrrhiza flavonoids and cytoxan had significant effect on the fluorescence pixels of RNA/DNA(P
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Objective To investigate the effects of cinnamaldehyde and citral on DNA and RNA of Aspergillus flavus and A. fumigatus cells and their mechanisms. Methods A. flavus and A. fumigatus were incubated on Czapeks agar plate (treated with cinnamaldehyde and citral at different concentrations) at 26.5 ℃ for 3—6 d. The normal and treated cells were observed by laser scanning confocal microscope (LSCM) and image analysis to describe the DNA and RNA levels by quantity and localization. Results DNA and RNA levels were changed greatly and multinucleate coniospores appeared in the treated cells. Conclusion Cinnamaldehyde and citral have directly or indirectly interfered the conventional synthesis of fungal hereditary DNA and RNA and normal differentiation of conidiophore in A. flavus and A. fumigatus, thus inhibiting the normal cell cycle and the growth and propagation of fungi.
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Objective To investigate the effects of laser irradiation on intracellular ROS(reactive oxidant species),intracellular calcium concentration(_i,and cell membrane integrity in the process of live cell imaging with confocal laser scanning microscopy. Methods The effects of a given laser irradiation on ROS,intracellular calcium concentration(_i and cell viability were revealed respectively by stained ECV-304 with H_2DCFDA,Fluo-4AM and calcein-AM/PI,and visualized and analyzed using ultra view LCI(live cell image)confocal microscopy. Results The irradiation of 488nm laser induced fluorescent intensity of DCF to increase abruptly and attain the climax in about 80 seconds,afterwards the fluorescent intensity fell and returned to the baseline.In the 70 minutes of the irradiation,the fluorescent intensity of intracellular Fluo-4 kept a slightly ascending tendency.The fluorescent intensity of calcein decreased 15minutes after the irradiation,and serval cells were PI positively stained.Conclusion 488nm laser irradiation induces intracellular reactive oxidant species(ROS) and calcium concentration to increase,but there is no significant influence on cell membrane integrity.