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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,
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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.
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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 .
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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 .
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10.3969/j.issn.1000-8179.2013.12.015
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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.
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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.
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<p><b>OBJECTIVE</b>To study the membrane mobility of aquaporin 7 (AQP7) by cloning stably transfected CHO cells with expression of pEGFP-C1-AQP7, in which AQP7 cDNA was fused downstream and in frame to pEGFP-C1 gene.</p><p><b>METHODS</b>The full sequence of AQP7 was amplified by RT-PCR and then recombined in the downstream of the green fluorescent protein gene in the pEGFP-C1 vector. The recombinant vector pEGFP-C1-AQP7 was stably transfected into CHO cells. With fluorescent microscopy, immunocytochemical stain and Western blot, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.</p><p><b>RESULTS</b>(1) The sequence of AQP7 cDNA of the Wistar rat was logged into the GenBank (access number: AY157737). (2) Identification demonstrated that pEGFP-C1-AQP7 fusion protein stably expressed in CHO cells. (3) With fluorescence microscopy, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.</p><p><b>CONCLUSION</b>The CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.</p>
Subject(s)
Animals , Cricetinae , Male , Rats , Aquaporins , Genetics , CHO Cells , Cricetulus , DNA, Complementary , Genetics , Green Fluorescent Proteins , Genetics , Molecular Sequence Data , Rats, Wistar , Recombinant Fusion Proteins , Reverse Transcriptase Polymerase Chain Reaction , Testis , Metabolism , TransfectionABSTRACT
AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate ［La(NO3)3］ and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO3)3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO3)3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO3)3 can depress the expression of AQP 7 in the rat testis.
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Aquaporins are small integral membrane proteins that selectively transport water and some small solutes in mammals, plants and lower organisms. Continued studies of the aquaporins are providing detailed tissue localization and the important physiological roles of aquaporins in urinary, respiratory, digestive and nervous systems. Based on the importance of aquaporin studies in the field of structural chemistry in biomembranes, American scientist Peter Agre won the 2003 Chemistry Nobel Prize for the discovery of water channels.
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Autosomal dominant polycystic kidney diseases are a large family of inherited diseases,which are characterized by the development of multiple renal cysts of tubular epithelial cell origin. Progressively enlarging cysts compromise normal renal parenchyma,reduce renal function and lead to renal failure. This review article summarizes recent literatures on the intracellular calcium homeostasis and signaling involving cAMP,EGFR and Ras/ERK,Wnt,m-TOR,as well as JAK-STAT,in the pathogenesis of polycystic kidney disease.
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Autosomal dominant polycystic kidney disease(ADPKD),a common inherited disease,is characterized by massive enlargement of fluid-filled renal cysts.Progressively enlarging cysts compromise normal renal parenchyma,reduce renal function and lead to renal failure.Up to now,the treatment options for ADPKD have been limited to renal replacement therapy by dialysis or by transplantation for patients with end-stage renal failure.Inhibition of cyst fluid secretion,suppression of cyst epithelial cell growth and prevention of renal failure are new approaches to treat PKD.
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AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate [La(NO 3) 3] and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO 3) 3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO 3) 3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO 3) 3 can depress the expression of AQP 7 in the rat testis.
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AIM:Changes of rat renal aquaporin (AQP1) and AQP2 mRNA expression after partially ligating left renal vein were observed to explore molecular mechanism of renal tubular reabsorption decrease resulted by increasing renal vein pressure.METHODS:Adult male rats were randomly divided into left renal vein partial ligation group(ie. VCG, varicocele group ) and sham operation group (SOG), left renal mRNA expression was analyed by Northen blot two months later.RESULTS:Intensity of left renal AQP1 mRNA expression in VCG was weaker than that in SOG, there was not any difference in left renal AQP2 mRNA expression between two groups, no hybridization signal was found in testes.CONCLUSION:Decrease of renal tubular reabsorption resulted by partially ligating renal vein might be related to decrease of AQP1 mRNA expression; “hormone escape" might occur in collecting tubules.
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The cDNA fragment of human c—fos oncogene has been obtained by a reverse transcription—DNA poly-merase chain reaction.Then the cDNA was ligated into pGEM3Zf(+)plasmid cut with Sma Ⅰ.with the re-combined plasmid,c—fos antisense RNA was synthesized in T7 RNA polymerase system.The primary appli-cation of anti-sense RNA is also studied in this article.