TO DEVELOP A NEW SYSTEM TO CONTIMUOUSLY MONITOR THE FUNCTIONS AT THE CELLULAR AND MOLECULAR LEVELS OF THE VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS IN VIVO.

1.血管内皮细胞和平滑肌细胞功能监测系统的研制：该系统具有以下特点：(1)利用步进电机驱动的光栅和斩波镜，可以用340 nm和380 nm激发光交替(10 Hz)照射组织样品，以测量细胞内钙离子浓度。(2)由研究人员设计的软件通过个人计算机控制斩波系统。(3)使用斩波镜和SIT相机获得荧光图像。显示火焰为640x400点和8位火焰。(4)在5 Hz频率下可获得荧光比值(F340/F380)图像。(5)可以在电视监视器上的图像中同时记录30个点的[Ca]i水平的变化。(6)系统制造成本低。利用该系统、前表面荧光法和分子生物学技术获得的新发现：(1)利用新发展的技术--Fura-2和二氨基萘的同时荧光法，我们在原位发现了内皮细胞内[Ca]i-NO的产生关系。(2)采用前表面荧光法、逆转录-聚合酶链式反应以及肾动脉和冠状动脉，我们发现肾上腺髓质素和内皮素在这些组织中都可能是自分泌/旁分泌的信号递质。Humana Press(新泽西州：美国)要求我们在一本名为《钙信号协议(DG Lambert编辑)》的书中写一章《使用正面荧光法测量肌条中的[Ca]i》，这本书是《分子生物学》系列方法的一部分。它正在印刷中。

1. THE DEVELOPMENT OF A NEW SYSTEM FOR MONITORING THE FUNCTIONS OF THE VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS : The system we have developed has following characteristics ; (1) Using a grating and a chopper mirror driven by a stepping motor, it became possible to alternately (10 Hz) illuminate the tissue sample with 340 and 380 nm excitation lights for the measurement of intracellular calcium concentration, [Ca] i. (2) the chopper system is controlled through a personal computer by investigator designed software. (3) Fluorescence images are obtained using a chopper mirror and a SIT camera. A display flame is 640x400 dots and a 8 bit flame. (4) Fluorescence ratio (F340/F380) images can be obtained at 5 Hz. (5) It is possible to simultaneously record changes in the [Ca] i levels at 30 points in an image on the TV monitor. (6) Low cost for making the system.2. NEW FINDINGS OBTAINED USING THIS SYSTEM,FRONT-SURFACE FLUORIMETRY AND TECHNIQUES IN MOLECULAR BIOLOGY : (1) Using the newly developed technique, the simultaneous fluorimetry of fura-2 and diaminonaphthalene, we found the [Ca] i-NO production relationships in the endothelial cells in situ. (2) Using front-sueface fluorimetry, reverse transcription polymerase chain reaction, and the renal and coronary arteries, we found that both adrenomedullin and endothelin might act as autocrine/paracrione signal transmitters in these tissues.3. HUMANA PRESS (NJ : U.S.A) asked us to write a chapter "Measurement of [Ca] i in muscle strips using front-surface fluorimetry" of a book on "Calcium signaling protocols (edited by DG Lambert)", a part of the Methods in Molecular Biology series. It is in press.