1.Study of the distribution and morphologic feature of the renal lymphatics
Taoyan ZHANG ; Gang LIU ; Guangju GUAN
Acta Universitatis Medicinalis Anhui 2013;(12):1449-1452
Objective To observe the fine distribution and morphologic feature of the intrarenal lymphatics. Meth-ods 20 Wistar rats were involved in this study and nephridial tissues were obtained from each animal. Double im-munofluorescence was applied to detect the lymphatic distribution by using Podoplanin and CD34. Renal tissues were obtained from the cortex, medulla and junctional zone,and put in 3% glutaric dialdehyde stationary liquid for transmission electron microscope to observe the fine distribution and morphologic feature of the intrarenal lymphat-ics. Results Under immunofluorescence double staining, lymphatic capillary appeared anomalism, large lumina, thin wall and located in connective tissue region. Cortilymph obviously exceeded medulla lymph in rats. Under e-lectron microscope, the basal lamina of lymphatic was discontinuity. A lot of lysosome and vesicles existed in the endothelial of capillary lymphatics. Many of the vesicles liberated in cytoplasm. The intercellular junctions of the renal lymphatics were held in close apposition. Conclusion The intercellular junctions of the renal lymphatics are held in close apposition by adhesion devices. A lot of lysosomes and vesicles exist in the endothelial of capillary lymphatics, which are the base for renal to transport big molecules and fluids.
2.Effect of heat shock protein 70 expression in Curcumin’s inhibition on STS-induced neurons injury
Zhe GUO ; Taoyan LIU ; Ruiyuan PAN ; Xiaoyan QIN
Chinese Journal of Biochemical Pharmaceutics 2014;37(4):24-27
Objective To explore the effect of heat shock protein 70(HSP70)expression in the role of Curcumin inhibited staurosporine(STS)-mediated neurons toxic injury.Methods The primary cultured hippocampal neurons was cultured in vitro and the stress damage model of STS-induced nerve cell toxicity was established.The experiment were divided into six groups according to the added drugs:normal control group,the STS model group (final concentration was 20μmol/L),Quercetin+STS model group(final concentration were 10 μmol/L and 20 μmol/L,respectively),Curcumin+STS pretreatment group(for 20μmol/L final concentration),Curcumin+Quercetin+STS treatment group(final concentration were 20μmol/L,10μmol/L and 20μmol/L,respectively)and Curcumin treatment group(final concentration was 20μmol/L).The cell viability were determined by thiazole blue (MTT)method,cell toxicity were measured by lactate dehydrogenase(LDH)release rate and HPS70 expression were detected by Western Blot. Results MTT results showed that the cell viability of Curcumin+STS pretreatment group was significantly higher than STS model group(P<0.001).Compared with Quercetin+STS model group,the cell viability of Curcumin+Quercetin+STS treatment group had little change.LDH results show that the nerve cell toxicity of Curcumin+STS pretreatment group was obviously less than that of STS model group(P<0.001).Western Blot results show that compared with STS model group,HSP70 protein expression in Curcumin+STS pretreatment group was significantly increased(P<0.001).Conclusion Curcumin can inhibit STS-mediated neurons toxicity stress damage though increasing HSP70 expression,when added Quercetin to block HSP70 expression in nerve cells,the inhibiting effect of Curcumin on STS-mediated neuron toxic stress injury is counteract.
3.Recombinant human serum albumin promotes differentiation of human pluripotent stem cells into cardiomyocytes in vitro
LIU Taoyan ; CUI Wei ; LI Hongxia ; WU Fujian ; LAN Feng
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2018;25(7):604-609
Objective To investigate whether recombinant human serum albumin (rHSA) can replace traditional B27 as a basic medium for differentiation of human pluripotent stem cells (hPSCs) into cardiomyocytes. Methods hPSCs were seeded at a cell density of 1.2×104/cm2; until up to 75% confluency hPSCs were induced by differentiation medium containing various concentration of rHSA (0, 50, 100, 200 g/L). Light microscope and fluorescence microscope recorded the whole process of stem cells differentiating into myocardium. Flow cytometry was used to detect the cardiac differentiation efficiency at different concentrations of rHSA. Immunofluorescence staining was used to detect the cardiac specific protein α-actinin and troponin T (cTnT) and electron microscope to observe the ultrastructure of human pluripotent stem cell-derived cardiomyocytes (hPSC-CM) and beating rates of hPSC-CMs response to drugs. Results A large number of spontaneous beating cardiomyocytes were observed 9 days after induction and differentition. The percentage of colonies showing beating cardiomyocytes was 60.4% at the concentration of 200 g/L of rice derived-rHSA. Beating cardiomyocytes were α-actinin and cTnT positive. Ultrastructural analysis showed scattered sarcomeres and mitochondrial. hPSC-CMs were dose-dependent on isopropyl adrenaline and verapamil. Conclusion Using such simple media to differentiate hPSCs into functional cardiomyocytes is cost-effective and highly efficient, and can be used in the clinical research.