The purpose of the present study was to determine the role of inositol 1,4,5-trisphosphate receptor 3 (IP₃R3) in renal cyst development in autosomal dominant polycystic kidney disease (ADPKD). 2-aminoethoxy-diphenyl borate (2-APB) and shRNA were used to suppress the expression of IP₃R3. The effect of IP₃R3 on cyst growth was investigated in Madin-Darby canine kidney (MDCK) cyst model, embryonic kidney cyst model and kidney specific Pkd1 knockout (PKD) mouse model. The underlying mechanism of IP₃R3 in promoting renal cyst development was investigated by Western blot and immunofluorescence staining. The results showed that the expression level of IP₃R3 was significantly increased in the kidneys of PKD mice. Inhibiting IP₃R3 by 2-APB or shRNA significantly retarded cyst expansion in MDCK cyst model and embryonic kidney cyst model. Western blot and immunofluorescence staining results showed that hyperactivated cAMP-PKA signaling pathway in the growth process of ADPKD cyst promoted the expression of IP₃R3, which was accompanied by a subcellular redistribution process in which IP₃R3 was translocated from endoplasmic reticulum to intercellular junction. The abnormal expression and subcellular localization of IP₃R3 further promoted cyst epithelial cell proliferation by activating MAPK and mTOR signaling pathways and accelerating cell cycle. These results suggest that the expression and subcellular distribution of IP₃R3 are involved in promoting renal cyst development, which implies IP₃R3 as a potential therapeutic target of ADPKD.
Animals ; Dogs ; Mice ; Cysts/genetics* ; Inositol 1,4,5-Trisphosphate Receptors/pharmacology* ; Kidney/metabolism* ; Polycystic Kidney Diseases/metabolism* ; Polycystic Kidney, Autosomal Dominant/drug therapy* ; Madin Darby Canine Kidney Cells

Urea transporters (UT) play a vital role in the mechanism of urine concentration and are recognized as novel targets for the development of salt-sparing diuretics. Thus, UT inhibitors are promising for development as novel diuretics. In the present study, a novel UT inhibitor with a diarylamide scaffold was discovered by high-throughput screening. Optimization of the inhibitor led to the identification of a promising preclinical candidate,

Urea transporters (UTs) are transmembrane urea-selective channel proteins that include two UT subfamilies, UT-A and UT-B. UT-A subfamily includes six members, UT-A1 to UT-A6, which are mainly expressed in kidney. UT-B subfamily has only one member that has a wide distribution in the body. UTs have been confirmed to play important roles in urinary concentration via the phenotypic analysis of 6 UT selective knockout mouse models. Experimental results suggest that UTs might be diuretic targets and that UT inhibitors might be developed as novel diuretics. This article reviews the physiological function and drug discovery of UT.
Animals ; Diuretics ; Kidney ; physiology ; Membrane Transport Proteins ; physiology ; Mice ; Mice, Knockout ; Urea

Thrombin activated fibrinolysis inhibitor ( TAFI) is a kind of plasma enzymes that can be activated by thrombin.TAFI regulates blood coagulation and fibrinolysis and has a strong fibrinolytic and anti-inflammatory ac-tivity.Its inhibitor is expected to minimize the risk of bleeding when thrombolytic recanalization .Also, studies found that TAFI played an important role in the development of cerebral thrombosis;atherosclerosis and so on .

Objective:To detect the expressions of urea transporter B (UT-B) in tumor tissue of bladder cancer patients and normal urothelium tissue, and to investigate its clinical significance of expression in bladder cancer tissue.Methods: Fifty-two paraffin-embedded specimens of bladder cancer patients and 15 normal urothelium specimens were selected. The expression levels of UT-B protein were detected by immunohistochemistry method.The difference of expression of UT-B in bladder cancer tissue and normal urothelium tissue and its relationship with the clinicopathological parameters were analyzed.Results:The expression rate of UT-B protein in bladder cancer tissue was 44.2%, while it was significantly higher in control group (93.3%), and the difference between two groups was significant (P=0.001).The positive expression rates of UT-B in bladder cancer tissue were significantly decreased with the increasing of histological grades, and there were significant differences between different grade groups (P=0.010).The positive expression rate of UT-B in muscle-invasive stage (Ta+T1) was significantly lower than that in non-muscle-invasive stage (T2+T3) (P=0.014).Conclusion:The reduction or lack of UT-B expression may promote the incidence, progression and invasiveness of bladder cancer.

Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .

10.3969/j.issn.1000-8179.2013.12.015

The function theory of an anesthesia vaporizer was studied and the geometry configuration was measured in this study. The internal gas flow and mixing process in the anesthesia vaporizer were simulated using CFD method. Applying tracking in turbulent flow to stochastic particle, for the droplet of anesthesia drug, the moving track of droplet was traced. Based on the results, the internal gas flow variation, the concentration distribution of anesthesia drug volatilization process and mixing process with gas were ascertained. Numerical simulation results showed that, the diluted gas velocity reduction of internal flow in the anesthesia vaporizer was higher. Because of the anesthesia vaporizer geometry, the mixing process between anesthesia drug vapor and diluting gas was not homogeneous. This also influenced the stability and accuracy of anesthesia drug concentration. The optimization precept of anesthesia vaporizer is ascertained.
Anesthesia, Inhalation ; instrumentation ; Computer Simulation ; Humans ; Imaging, Three-Dimensional ; Nebulizers and Vaporizers ; standards ; Numerical Analysis, Computer-Assisted

Natural products were the material basis of drug discovery and drug screening, and high-performance techniques were the important prerequisite of earning hits. In this paper, we reviewed the current situation and future prospects of drug screening, including the pharmaceutical environment, the challenges facing drug discovery, the screening tools and the methods of generating analogs.

Objective To study the expression and regulation of aquapofins (AQP) in cystic epithelial cells of jck mice with polycystic kidney disease. Methods Localization and regulation of AQP1, AQP2, AQP3 and AQP4 protein were analyzed by using the immunofluorescence and Western blotting. Results Kidneys of jck homozygous mice were 4 folds larger than those of litter matched wild-type mice. There were multiple cysts and fibrosis in the renal tissue of jck mice. The epithelial cells in cysts were flat in shape. Blood urea level in jck mice was (42.6 ± 6.7) mmol/L, which was 5 folds higher than that in wild-type mice [(8.4±1.9) mmol/L] (P<0.01). Immunofluorescence analysis showed that AQP1 was expressed in the apical and hasolatend membranes of epithelial cells in proximal tubules, as well as in the thin descending limb of Henle and endothelial cells of descending vasa recta. There was no AQP1 expression in epithelial cells of cysts. AQP2 was expressed in the apical membranes of collecting ducts and renal cysts. AQP3 and AQP4 were expressed in basolateral membranes of collecting duct and renal cystic epithelial cells of jck mice. Western blot analysis showed the same protein sizes of AQP1, AQP2, AQP3 and AQP4 in both jck and wild-type kidneys. However, AQP1 expression was down-regulated in jck kidneys (P<0.01). Conclusion The renal cystic epithelia expresses AQP2, AQP3 and AQP4, which indicates that epithelial cells in renal cysts are derived from renal collecting ducts in jck mice and aquaporins may play an important role in renal cyst development.