| Objective Based on the platelet are in activation state in many diseases, but there is not a method capable of detecting a comprehensive response to the complexity of the platelet biology and function before and after the antiplatelet drugs. This study would exploringly apply UV, IR and soft X-ray spectroscopic imaging microscopy to the detection of platelet aggregation and to observe the changes of platelets in coronary heart disease patients with normal platelet, so as to realize the impact of anticoagulants on platelet function.Methods Research was classified into clinical and basic parts. In clinical study, 20 elderly patients with coronary artery disease randomly selected, 10 healthy elderly volunteers were similarity chosen as a control group. Being informed and extracted 2ml blood using the second centrifugation for platelet-rich plasma. We took UV spectrophotometer to obtain ultraviolet spectra of the platelet between the experiment group and control group, and then software UVWIN5.0 was used for analyze. And then another 10 healthy subjects was analyzed by infrared spectroscopy using liquid method for preliminary exploratory research. In the basic research, 10 male SD rats were selected and divided as clopidogrel group and control group. As the platelet being extracted, it’s been invested adopting soft X-ray spectroscopy and microscopy techniques to watch the calcium distribution in platelet by dual-energy contrast imaging and the modalities of platelet by scanning transmission imaging. Theoutcome was plotted using Origin8.0 and the statistical data processed using SPASS13.0.Results1. The ultraviolet spectroscopy of platelet had a absorption peak at 204 nm and280nm wavelengths, while the absorption was significantly greater after aggregation than un-aggregation at 204 nm wavelength( clopidogrel group,t=3.475, P=0.007; aspirin group, t=4.593, P=0.001; control group, t=5.003,P=0.001). The position of spectrum had no apparent distinction between the coronary heart disease group and healthy group which appeared at the point of204 nm and 280 nm.2. The infrared spectroscopy had been appearing three peaks of wave. They appeared at wavelength of 1550cm-1 which represents hydrogen bond bending vibration,2360 cm-1 presents CO2 vibration, 2930cm-1 and 3300cm-1 separately stand for single bond stretching vibration. Although the infrared absorbance increased after gathering, it’s related with the distribution of platelet in the Ca F2 substrate and there had no new peak of wave appearing.3. Using the soft X-ray scanning technology we could see the morphology of varied shape and size, even the platelet pseudopodia could been expressed, but the resolution was limited by the definition. The dual-energy contrast imaging showed the distribution of intracellular calcium in platelet mainly concentrated in the calcium pool at the side of plasma membrane in resting state, while distributed throughout the cytoplasm after induced. Not only the scanning transmission imaging but also the dual-energy contrast imaging don’t have statistical differencecause influenced by the inducer and the factor external in both normal and clopigredel group.Conclusion Distinction of platelet aggregation and activity could be discovered in a variety of physical methods which can be applied to provide the basis for its detection.Dual-beam ultraviolet spectrometer and an infrared spectrometer could make ultraviolet and infrared spectrum that can show the difference agility and accurately,while the infrared spectrum had superiority to revealing the peculiar functional group which could reveal the internal molecular structure. Moreover, the soft X-ray microscopic imaging provided a new method for the detection in natural state and the spatial distribution of imaging and quantitative analysis. |
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