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Objective To establish the spatial course of distal tubule and afferent arterioles after macula densa, and to locate and detect the proteins in the adjacent parts by using three-dimensional visualization technology of microstructure. Methods C57 BL/6J mice were fixed by perfusion and embedded in epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney. And a total of 720, 2. 5 μm-thick consecutive sections were obtained from the renal capsule to the outer stripe of the renal outer medulla. After aligning the digital microscopic images through computer registration procedures, the tubules and vessels were traced by 3D reconstruction program edited by C Language. Selecting the tissue sections of the contact site and applying the improved immunoperoxidase staining method to detect H
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Objective To describe quantitatively the development of the capillary loop stage glomerulus (capG) with respect to the volume density of capillaries in the glomerulus based on the morphogenesis of the kidney. Methods The kidneys were obtained from mice at various developing time points and prepared for paraffin sections. The volume density of CD34 positive endothelial cells and surrounded capillary lumen in glomeruli was measured using a combination of immunohistochemical staining and the stereological grid system. Results The capG was divided into early, middle, and late phases, and middle phase capG was subdivided into early-middle and late-middle phases, according to the morphology of developing glomeruli and the arrangement of podocytes. As result, the volume density of capillary loops in early phase capG could not be measured due to the complex "glomerular" shape. The volume density of capillary loops increased from (35.95±6.45)% in the early-middle phase capG, to (58.36±6. 30) % in the late-middle phase capG, and to (79.89± 5.21) % in the late phase capG, compared to (93.61 ±1.96) % in the mature glomerulus. Furthermore, the volume density of capillary loops remained constant at same stage even though at different developmental time points. Conclusion This study demonstrated a significantly increased volume density of capillary loops with the kidney development. In addition, the results provide a descriptive and reliable parameter for the evaluation of glomerular development.
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Objective:To observe the effect of Chaibei Zhixian decoction and peimine on Carbamazepine (CBZ) concentration, P-glycoprotein (P-gp) and multi drug resistance 1(MDR1) expression in the brain tissues of rats with refractory epilepsy, and to understand the contribution of Peimine in the compound prescription to treat the refractory epilepsy. Method:Epilepsy rat models were established by injecting kainic acid (KA) in the lateral ventricle. The successfully modeled rats were randomly divided into model group, CBZ group(0.12 g·kg-1),Chaibei Zhixian decoction+CBZ group(8.39 g·kg-1+0.12 g·kg-1), peimine+CBZ group(0.01 g·kg-1+0.12 g·kg-1) and sham operation group. After 60 days of intervention, the expression levels of P-glycoprotein (P-gp) and MDR1b mRNA in the brain cortex were detected by Western blot and quantitative real\|time fluorescence polymerase chain reaction(Real-time PCR),the contents of CBZ and 10,11-epoxidation of carbamazepine (CBZE) were measured by liquid chromatography-mass spectrometry (LC-MS). Result:Compared with sham group, the expression of P-gp/MDR1 in the cortex of model group was significantly increased (PPPPPPPConclusion:Chaibei Zhixian decoction and peimine may increase the content of CBZ and CBZE in the brain tissues in rats with intractable epilepsy by reducing the expression of MDR1/P-gp in the cortex.