To investigate the relation of two different mutations to the outcome of partial external biliary diversion (PEBD) in severe bile salt export pump (BSEP) deficiency. METHODSMutations in the gene encoding BSEP leading ...To investigate the relation of two different mutations to the outcome of partial external biliary diversion (PEBD) in severe bile salt export pump (BSEP) deficiency. METHODSMutations in the gene encoding BSEP leading to severe BSEP deficiency in two unrelated patients were identified by genomic sequencing. Native liver biopsies and transiently transfected human embryonic kidney (HEK) 293 cells expressing either wild-type or mutated BSEP were subjected to immunofluorescence analysis to assess BSEP transporter localization. Bile acid profiles of patient and control bile samples were generated by ultra-performance liquid chromatography-tandem mass spectrometry. Wild-type and mutant BSEP transport of [<sup>3</sup>H]-labeled taurocholate (TC) and taurochenodeoxycholate (TCDC) was assessed by vesicular transport assays. RESULTSA girl (at 2 mo) presented with pruritus, jaundice and elevated serum bile salts (BS). PEBD stabilized liver function and prevented liver transplantation. She was heterozygous for the BSEP deletion p.T919del and the nonsense mutation p.R1235X. At the age of 17 years relative amounts of conjugated BS in her bile were normal, while total BS were less than 3% as compared to controls. An unrelated boy (age 1.5 years) presenting with severe pruritus and elevated serum BS was heterozygous for the same nonsense and another missense mutation, p.G1032R. PEBD failed to alleviate pruritus, eventually necessitating liver transplantation. BS concentration in bile was about 5% of controls. BS were mainly unconjugated with an unusual low amount of chenodeoxycholate derivatives (< 5%). The patients’ native liver biopsies showed canalicular BSEP expression. Both BSEP p.T919del and p.G1032R were localized in the plasma membrane in HEK293 cells. In vitro transport assays showed drastic reduction of transport by both mutations. Using purified recombinant BSEP as quantifiable reference, per-molecule transport rates for TC and TCDC were determined to be 3 and 2 BS molecules per wild-type BSEP transporter per minute, respectively. CONCLUSIONIn summary, our findings suggest that residual function of BSEP as well as substrate specificity influence the therapeutic effectiveness of PEBD in progressive familial intrahepatic cholestasis type 2 (PFIC-2).展开更多
Objective： To explore the electrophysiological proper- ties of differentiation of rat bone marrow-derived stromal stem cells （rBMSCs） to neuron-like cells in vitro by edaravone, a new type of free radical scavenger...Objective： To explore the electrophysiological proper- ties of differentiation of rat bone marrow-derived stromal stem cells （rBMSCs） to neuron-like cells in vitro by edaravone, a new type of free radical scavenger. Methods： Stromal stem cells were separated from rat bone marrow with Ficoll-Paque reagent and expanded in different culture medium in vitro, rBMSCs were induced by edaravone containing serum-free L-DMEM. Morphologic observation and Western blot analysis including the expression of Nav1.6, Kvl.2, Kv1.3, Cav1.2 were performed, and whole patch-clamp technique was used. Results： Cyton contraction and long processes were shown in differentiated stromal stem cells. Navl.6, Kvl.2, Kv 1.3 and Cav 1.2 were expressed in both differentiated and undifferentiated ceils. However, the expression of channel proteins in differentiated cells was up-regulated. Consistently, their resting potential and outward currents were also enhanced in the differentiated cells, which was especially significant in the outward rectifier potassium current. Conclusion： In vitro, neuron-like cells derived from rBMSCs, induced by edaravone, possess electrophysiological properties of neurons.展开更多
Herein we report the room-temperature epitaxial growth of V203 films by laser molecule beam epitaxy. X-ray diffraction pro- files show the room-temperature epitaxial V2O3 films orient in the [ 110] direction on α-Al2...Herein we report the room-temperature epitaxial growth of V203 films by laser molecule beam epitaxy. X-ray diffraction pro- files show the room-temperature epitaxial V2O3 films orient in the [ 110] direction on α-Al2O3 （0001） substrates. Atomic force microscopy measurements reveal that the ultra-smooth surfaces with root-mean-square surface roughness of 0.11 nm and 0.28 nm for 10-nm-thick and 35-nm-thick V2O3 film, respectively. X-ray photoelectron spectroscopy results indicate the V3＋ oxida- tion state in the films. Typical metal-insulator transition is observed in films at about 135 K. The resistivities at 300 K are ap- proximately 0.8 mΩ cm and 0.5 mΩ cm for 10-rim-thick and 35-nm-thick V203 film, respectively.展开更多
基金Supported by the German -Research Foun-dation-through the Clin-ical Research Group KFO217“Hepatobiliary tran-sport an-d liver diseases”the Collaborative Research Cen-tre 974“Commun-ication-an-d Systemic Relevan-ce in-Liver Damage an-d Regen-eration-”
文摘To investigate the relation of two different mutations to the outcome of partial external biliary diversion (PEBD) in severe bile salt export pump (BSEP) deficiency. METHODSMutations in the gene encoding BSEP leading to severe BSEP deficiency in two unrelated patients were identified by genomic sequencing. Native liver biopsies and transiently transfected human embryonic kidney (HEK) 293 cells expressing either wild-type or mutated BSEP were subjected to immunofluorescence analysis to assess BSEP transporter localization. Bile acid profiles of patient and control bile samples were generated by ultra-performance liquid chromatography-tandem mass spectrometry. Wild-type and mutant BSEP transport of [<sup>3</sup>H]-labeled taurocholate (TC) and taurochenodeoxycholate (TCDC) was assessed by vesicular transport assays. RESULTSA girl (at 2 mo) presented with pruritus, jaundice and elevated serum bile salts (BS). PEBD stabilized liver function and prevented liver transplantation. She was heterozygous for the BSEP deletion p.T919del and the nonsense mutation p.R1235X. At the age of 17 years relative amounts of conjugated BS in her bile were normal, while total BS were less than 3% as compared to controls. An unrelated boy (age 1.5 years) presenting with severe pruritus and elevated serum BS was heterozygous for the same nonsense and another missense mutation, p.G1032R. PEBD failed to alleviate pruritus, eventually necessitating liver transplantation. BS concentration in bile was about 5% of controls. BS were mainly unconjugated with an unusual low amount of chenodeoxycholate derivatives (< 5%). The patients’ native liver biopsies showed canalicular BSEP expression. Both BSEP p.T919del and p.G1032R were localized in the plasma membrane in HEK293 cells. In vitro transport assays showed drastic reduction of transport by both mutations. Using purified recombinant BSEP as quantifiable reference, per-molecule transport rates for TC and TCDC were determined to be 3 and 2 BS molecules per wild-type BSEP transporter per minute, respectively. CONCLUSIONIn summary, our findings suggest that residual function of BSEP as well as substrate specificity influence the therapeutic effectiveness of PEBD in progressive familial intrahepatic cholestasis type 2 (PFIC-2).
基金The work was supported by Natural Science Foundation of Guangdong Province （No.06028967, 8452402301001081 ）.
文摘Objective： To explore the electrophysiological proper- ties of differentiation of rat bone marrow-derived stromal stem cells （rBMSCs） to neuron-like cells in vitro by edaravone, a new type of free radical scavenger. Methods： Stromal stem cells were separated from rat bone marrow with Ficoll-Paque reagent and expanded in different culture medium in vitro, rBMSCs were induced by edaravone containing serum-free L-DMEM. Morphologic observation and Western blot analysis including the expression of Nav1.6, Kvl.2, Kv1.3, Cav1.2 were performed, and whole patch-clamp technique was used. Results： Cyton contraction and long processes were shown in differentiated stromal stem cells. Navl.6, Kvl.2, Kv 1.3 and Cav 1.2 were expressed in both differentiated and undifferentiated ceils. However, the expression of channel proteins in differentiated cells was up-regulated. Consistently, their resting potential and outward currents were also enhanced in the differentiated cells, which was especially significant in the outward rectifier potassium current. Conclusion： In vitro, neuron-like cells derived from rBMSCs, induced by edaravone, possess electrophysiological properties of neurons.
基金supported by the National Basic Research Program of China(Grant Nos.2010CB630704 and 2012CB921403)
文摘Herein we report the room-temperature epitaxial growth of V203 films by laser molecule beam epitaxy. X-ray diffraction pro- files show the room-temperature epitaxial V2O3 films orient in the [ 110] direction on α-Al2O3 （0001） substrates. Atomic force microscopy measurements reveal that the ultra-smooth surfaces with root-mean-square surface roughness of 0.11 nm and 0.28 nm for 10-nm-thick and 35-nm-thick V2O3 film, respectively. X-ray photoelectron spectroscopy results indicate the V3＋ oxida- tion state in the films. Typical metal-insulator transition is observed in films at about 135 K. The resistivities at 300 K are ap- proximately 0.8 mΩ cm and 0.5 mΩ cm for 10-rim-thick and 35-nm-thick V203 film, respectively.
