AIM:To examine whether heme oxygenase (HO)-1 overexpression would exert direct or indirect effects on Kupffer cells activation, which lead to aggravation of reperfusion injury.METHODS: Donors were pretreated with coba...AIM:To examine whether heme oxygenase (HO)-1 overexpression would exert direct or indirect effects on Kupffer cells activation, which lead to aggravation of reperfusion injury.METHODS: Donors were pretreated with cobalt protoporphyrin (CoPP) or zinc protoporphyrin (ZnPP), HO-1 inducer and antagonist, respectively. Livers were stored at 4℃ for 24 h before transplantation. Kupffer cells were isolated and cultured for 6 h after liver reperfusion.RESULTS: Postoperatively, serum transaminases were significantly lower and associated with less liver injury when donors were pretreated with CoPP, as compared with the ZnPP group. Production of the cytokines tumor necrosis factor-α and interleukin-6 generated by Kupffer cells decreased in the CoPP group. The CD14 expression levels (RT-PCR/Western blots) of Kupffer cells from CoPP-pretreated liver grafts reduced.CONCLUSION: The study suggests that the potential utility of HO-1 overexpression in preventing ischemia/reperfusion injury results from inhibition of Kupffer cells activation.展开更多
BACKGROUND: The non-function and dysfunction of primary liver graft likely involves dependence on Kupffer cells and hepatic innervation. The present experiment was designed to study the expression of P-selectin and in...BACKGROUND: The non-function and dysfunction of primary liver graft likely involves dependence on Kupffer cells and hepatic innervation. The present experiment was designed to study the expression of P-selectin and intercellular adhesion molecule-1 (ICAM-1) mRNA in liver graft and to elucidate the role of Kupffer cells and the sympathetic nerve of the liver in down-regulating this expression. METHODS: Donor rats were given hexamethonium, a sympathetic ganglionic blocking agent, and/or gadolinium chloride, an inhibitor of Kupffer cells. Then the changes of graft P-selectin and ICAM-1 mRNA expression were measured after liver transplantation. RESULTS: The expressions of P-selectin and ICAM-1 mRNA were increased after liver transplantation, and down-regulated by liver denervation and elimination of Kupffer cells. CONCLUSIONS: Live donor denervation and elimination of Kupffer cells down-regulated the expressions of P-selectin and ICAM-1 mRNA in grafts. This may decrease graft ischemia/reperfusion injury.展开更多
BACKGROUND: The mda-7/IL-24 receptor belongs to the type 11 cytokine receptor family, and its two heterodimeric receptors are IL-22R1/IL-20R2 and IL-20R1/IL-20R2. Mda-7/IL-24 receptor expression in liver cancer cell l...BACKGROUND: The mda-7/IL-24 receptor belongs to the type 11 cytokine receptor family, and its two heterodimeric receptors are IL-22R1/IL-20R2 and IL-20R1/IL-20R2. Mda-7/IL-24 receptor expression in liver cancer cell lines has not yet been described. This information may be helpful for further clinical gene therapy. METHODS: With normal skin total RNA as template, the cDNA sequences of IL-20R1, IL-20R2 and IL-22R were amplified by RT-PCR. Total RNA was extracted from cultured liver cancer cell lines and a normal liver cell line, then detected by northern blotting, and the expression of mda-7/IL-24 receptors was analyzed. RESULTS: PLC/PRF/5 and SMMC-7721 expressed IL-20R1; BEL-7402, Hep3B, HepG2, and PLC/PRF/5 expressed IL20R2; and HepG2 and PLC/PRF/5 expressed IL-22R. Only HepG2 expressed the IL-22R/IL-20R2 receptor complex. PLC/PRF/5 completely expressed both heterodimeric receptors. Huh-7, QGY-7701 and WRL-68 did not express the IL-24 receptor. CONCLUSION: Complete mda-7/IL-24 receptors are seldom expressed in liver cancer cell lines.展开更多
BACKGROUND: A suitable perfusate is very important in reducing various problems in liver preservation, prolonging the time of organ preservation and enhancing the quality of donor tissue. University of Wisconsin (UW) ...BACKGROUND: A suitable perfusate is very important in reducing various problems in liver preservation, prolonging the time of organ preservation and enhancing the quality of donor tissue. University of Wisconsin (UW) solution is the most successful solution for preserving multiple organs at present, but it has many shortcomings. We set out to develop a new liver preservation solution (KYL solution) and study its effects on apoptosis in rat liver undergoing cold preservation. METHODS: Using non-circulated isolated perfused rat liver (IPRL), we randomly preserved Sprague-Dawley rat livers for 0, 4, 8, 16, 24, and 48 hours with KYL solution or UW solution. The effects were assessed by measuring the content of free radicals in Krebs-Henseleit solution and the intracellular calcium content of hepatocytes, assessing hepatocellular apoptosis and related-gene expression, and observing the morphological changes in liver. To evaluate the protection by KYL and UW solutions in rat liver perfusion and preservation, we chosed normal saline for negative comparison. RESULTS: The intracellular calcium content of the liver preserved in KYL solution was less than that preserved in UW solution. At every different period of preservation, the malonaldehyde and superoxide dismutase content in Krebs-Henseleit solution, the percentage of apoptotic cells and the expression patterns of apoptosis-related-genes were similar in livers preserved in KYL and UW solutions. Morphological changes in the two groups were almost the same. The variables in both groups were better than those of livers preserved in normal saline. Both KYL and UW solutions protected rat liver from ischemia-reperfusion injury. CONCLUSIONS: KYL solution is superior to UW solution in preventing calcium overload. More severe hepatocyte damage may appear in the KYL group than in the UW group and the effect of KYL solution on apoptosis in rat liver preservation is similar to that of UW solution.展开更多
Background Intravascular microbubble-enhanced acoustic cavitation is capable of disrupting the vascular walls of capillaries and small vessels. This study was designed to investigate the impact of microbubble-enhanced...Background Intravascular microbubble-enhanced acoustic cavitation is capable of disrupting the vascular walls of capillaries and small vessels. This study was designed to investigate the impact of microbubble-enhanced, pulsed and focused ultrasound （MEUS） on the blood perfusion of subcutaneous VX2 tumors in rabbits. Methods Subcutaneous VX2 cancers in twenty New Zealand rabbits were treated by combining high-pressure amplitude, pulsed and focused therapeutic ultrasound （TUS） and intravenous microbubble injections. The TUS transducer was operated with a peak negative pressure of 4.6 MPa and a duty cycle of 0.41%. Controls were subcutaneous VX2 cancers treated with TUS or microbubbles only. Contrast-enhanced ultrasound （CEUS） and intravenous Evans Blue （EB） perfusion were performed to assess the tumor circulation. The tumor microvascular disruption was assessed by histological examination. Results CEUS showed that the tumor circulation almost vanished after MEUS treatment. The average peak grayscale value （GSV） of tumor CEUS dropped significantly from 84.1±22.4 to 15.8±10.8 in the MEUS-treated tumors but no significant GSV changes were found in tumors in the two control groups. The mean tumor EB content of the MEUS-treated tumors was significantly lower than that of the controls. Histological examination found scattered tumor microvascular disruption with intercellular edema after MEUS treatment. Conclusion The tumor circulation of VX2 cancers can be arrested or significantly reduced by MEUS due to microvascular disruption. Chin M~.cl ,I 2014：127 （14）： 2605-2611展开更多
Background Cardiac resynchronization therapy （CRT） with biventricular pacing improves cardiac function,functional capacity and quality of life in selected patients with heart failure.The current study aimed to evalu...Background Cardiac resynchronization therapy （CRT） with biventricular pacing improves cardiac function,functional capacity and quality of life in selected patients with heart failure.The current study aimed to evaluate the efficacy of the intracardiac electrogram （IEGM）-based optimization method,QuickOptTM,in Chinese patients treated with CRT.Methods Aortic time velocity integrals （AVTI） achieved at the sensed atrioventricular （AV）,paced AV and interventricular （VV） interval settings recommended by both QuickOptTM and standard echocardiographic optimization were measured in 101 patients.Consistency and the strength of the relationship between the two timing cycle optimization methods were assessed by intra-class correlation coefficient （ICC）.Results The ICC showed good agreement and correlation with what the AVTI achieved at the optimal sensed AV （ICC=0.9683 （0.9535-0.9785））,paced AV （ICC=0.9642 （0.9475-0.9757）） and VV （ICC=0.9730 （0.9602-0.9817）） interval settings determined by the two optimization methods.The average time required by echocardiographic optimization and by QuickOptTM were （78.32±32.40） minutes and （1.98±1.64） minutes respectively （P 〈0.0001）.Conclusion The QuickOptTM algorithm provides a quicker,simpler and reliable alternative to the standard method for timing cycle optimization.（ClinicaITrial.gov Reference Number：NCT00918294）展开更多
Two new ent-kaurene diterpenoids, angustifolin B and natural hydrogen-bonded 1:1 complex, angustifolin C, as well as four known diterpenoids, angustifolin, isodonal, sodoponin, tri-chorabdal B, were isolated from the ...Two new ent-kaurene diterpenoids, angustifolin B and natural hydrogen-bonded 1:1 complex, angustifolin C, as well as four known diterpenoids, angustifolin, isodonal, sodoponin, tri-chorabdal B, were isolated from the leaves of Rabdosia angustifolia.展开更多
Background Idiopathic hyperoxaluria （IH） may be caused by increased endogenous formation or exogenous absorption of oxalic acid.Characterization of the molecular pathogenesis of IH has been hampered by the lack of a...Background Idiopathic hyperoxaluria （IH） may be caused by increased endogenous formation or exogenous absorption of oxalic acid.Characterization of the molecular pathogenesis of IH has been hampered by the lack of an ideal animal model.We therefore established a stabile rat IH model in order to analyze variation in gene expression profile in the jejunum and to investigate the association between IH pathogenesis and exogenous absorption of oxalic acid.Methods A rat model of IH was established and three female rats with IH were assigned to the study group,while three normal rats served as controls.Total RNA was isolated from the jejunum of rats in the two groups and mRNA was purified,reversely transcribed,labeled with Cy5 or Cy3 and hybridized to 27K Rat Genome Array.Differences in gene expression profile between the 2 groups were analyzed by bioinformatics methods.Results Comparative analysis revealed that the expression of 517 genes was up-regulated and that of 203 genes was down-regulated by at least two-fold in the jejunum of rats with idiopathic hyperoxaluria.These genes are related to many functions including cell signal transduction,DNA binding and transcription,ATP binding,ion binding and transport,cell receptors,immunity,cyclins,cytoskeleton structure,and metabolic proteins.Kyoto encyclopedia of genes and genomes （KEGG） signaling pathway analysis revealed that the variations of 239 pathway functional changes are statistically significant （P 〈0.05）.Conclusions cDNA microarray can be used effectively to screen differentially expressed genes in the jejunum of rats with idiopathic hyperoxaluria. These differentially expressed genes may underlie idiopathic hyperoxaluria pathophysiology and provide a platform for further studying molecular pathogenetic mechanisms.展开更多
基金Supported by The Natural Science Foundation of Yunnan Province,China, No.2007C137Mthe Joint Funds of Natural Science Foundation of Yunnan Province,China,No.2007C0009R
文摘AIM:To examine whether heme oxygenase (HO)-1 overexpression would exert direct or indirect effects on Kupffer cells activation, which lead to aggravation of reperfusion injury.METHODS: Donors were pretreated with cobalt protoporphyrin (CoPP) or zinc protoporphyrin (ZnPP), HO-1 inducer and antagonist, respectively. Livers were stored at 4℃ for 24 h before transplantation. Kupffer cells were isolated and cultured for 6 h after liver reperfusion.RESULTS: Postoperatively, serum transaminases were significantly lower and associated with less liver injury when donors were pretreated with CoPP, as compared with the ZnPP group. Production of the cytokines tumor necrosis factor-α and interleukin-6 generated by Kupffer cells decreased in the CoPP group. The CD14 expression levels (RT-PCR/Western blots) of Kupffer cells from CoPP-pretreated liver grafts reduced.CONCLUSION: The study suggests that the potential utility of HO-1 overexpression in preventing ischemia/reperfusion injury results from inhibition of Kupffer cells activation.
文摘BACKGROUND: The non-function and dysfunction of primary liver graft likely involves dependence on Kupffer cells and hepatic innervation. The present experiment was designed to study the expression of P-selectin and intercellular adhesion molecule-1 (ICAM-1) mRNA in liver graft and to elucidate the role of Kupffer cells and the sympathetic nerve of the liver in down-regulating this expression. METHODS: Donor rats were given hexamethonium, a sympathetic ganglionic blocking agent, and/or gadolinium chloride, an inhibitor of Kupffer cells. Then the changes of graft P-selectin and ICAM-1 mRNA expression were measured after liver transplantation. RESULTS: The expressions of P-selectin and ICAM-1 mRNA were increased after liver transplantation, and down-regulated by liver denervation and elimination of Kupffer cells. CONCLUSIONS: Live donor denervation and elimination of Kupffer cells down-regulated the expressions of P-selectin and ICAM-1 mRNA in grafts. This may decrease graft ischemia/reperfusion injury.
文摘BACKGROUND: The mda-7/IL-24 receptor belongs to the type 11 cytokine receptor family, and its two heterodimeric receptors are IL-22R1/IL-20R2 and IL-20R1/IL-20R2. Mda-7/IL-24 receptor expression in liver cancer cell lines has not yet been described. This information may be helpful for further clinical gene therapy. METHODS: With normal skin total RNA as template, the cDNA sequences of IL-20R1, IL-20R2 and IL-22R were amplified by RT-PCR. Total RNA was extracted from cultured liver cancer cell lines and a normal liver cell line, then detected by northern blotting, and the expression of mda-7/IL-24 receptors was analyzed. RESULTS: PLC/PRF/5 and SMMC-7721 expressed IL-20R1; BEL-7402, Hep3B, HepG2, and PLC/PRF/5 expressed IL20R2; and HepG2 and PLC/PRF/5 expressed IL-22R. Only HepG2 expressed the IL-22R/IL-20R2 receptor complex. PLC/PRF/5 completely expressed both heterodimeric receptors. Huh-7, QGY-7701 and WRL-68 did not express the IL-24 receptor. CONCLUSION: Complete mda-7/IL-24 receptors are seldom expressed in liver cancer cell lines.
文摘BACKGROUND: A suitable perfusate is very important in reducing various problems in liver preservation, prolonging the time of organ preservation and enhancing the quality of donor tissue. University of Wisconsin (UW) solution is the most successful solution for preserving multiple organs at present, but it has many shortcomings. We set out to develop a new liver preservation solution (KYL solution) and study its effects on apoptosis in rat liver undergoing cold preservation. METHODS: Using non-circulated isolated perfused rat liver (IPRL), we randomly preserved Sprague-Dawley rat livers for 0, 4, 8, 16, 24, and 48 hours with KYL solution or UW solution. The effects were assessed by measuring the content of free radicals in Krebs-Henseleit solution and the intracellular calcium content of hepatocytes, assessing hepatocellular apoptosis and related-gene expression, and observing the morphological changes in liver. To evaluate the protection by KYL and UW solutions in rat liver perfusion and preservation, we chosed normal saline for negative comparison. RESULTS: The intracellular calcium content of the liver preserved in KYL solution was less than that preserved in UW solution. At every different period of preservation, the malonaldehyde and superoxide dismutase content in Krebs-Henseleit solution, the percentage of apoptotic cells and the expression patterns of apoptosis-related-genes were similar in livers preserved in KYL and UW solutions. Morphological changes in the two groups were almost the same. The variables in both groups were better than those of livers preserved in normal saline. Both KYL and UW solutions protected rat liver from ischemia-reperfusion injury. CONCLUSIONS: KYL solution is superior to UW solution in preventing calcium overload. More severe hepatocyte damage may appear in the KYL group than in the UW group and the effect of KYL solution on apoptosis in rat liver preservation is similar to that of UW solution.
文摘Background Intravascular microbubble-enhanced acoustic cavitation is capable of disrupting the vascular walls of capillaries and small vessels. This study was designed to investigate the impact of microbubble-enhanced, pulsed and focused ultrasound （MEUS） on the blood perfusion of subcutaneous VX2 tumors in rabbits. Methods Subcutaneous VX2 cancers in twenty New Zealand rabbits were treated by combining high-pressure amplitude, pulsed and focused therapeutic ultrasound （TUS） and intravenous microbubble injections. The TUS transducer was operated with a peak negative pressure of 4.6 MPa and a duty cycle of 0.41%. Controls were subcutaneous VX2 cancers treated with TUS or microbubbles only. Contrast-enhanced ultrasound （CEUS） and intravenous Evans Blue （EB） perfusion were performed to assess the tumor circulation. The tumor microvascular disruption was assessed by histological examination. Results CEUS showed that the tumor circulation almost vanished after MEUS treatment. The average peak grayscale value （GSV） of tumor CEUS dropped significantly from 84.1±22.4 to 15.8±10.8 in the MEUS-treated tumors but no significant GSV changes were found in tumors in the two control groups. The mean tumor EB content of the MEUS-treated tumors was significantly lower than that of the controls. Histological examination found scattered tumor microvascular disruption with intercellular edema after MEUS treatment. Conclusion The tumor circulation of VX2 cancers can be arrested or significantly reduced by MEUS due to microvascular disruption. Chin M~.cl ,I 2014：127 （14）： 2605-2611
文摘Background Cardiac resynchronization therapy （CRT） with biventricular pacing improves cardiac function,functional capacity and quality of life in selected patients with heart failure.The current study aimed to evaluate the efficacy of the intracardiac electrogram （IEGM）-based optimization method,QuickOptTM,in Chinese patients treated with CRT.Methods Aortic time velocity integrals （AVTI） achieved at the sensed atrioventricular （AV）,paced AV and interventricular （VV） interval settings recommended by both QuickOptTM and standard echocardiographic optimization were measured in 101 patients.Consistency and the strength of the relationship between the two timing cycle optimization methods were assessed by intra-class correlation coefficient （ICC）.Results The ICC showed good agreement and correlation with what the AVTI achieved at the optimal sensed AV （ICC=0.9683 （0.9535-0.9785））,paced AV （ICC=0.9642 （0.9475-0.9757）） and VV （ICC=0.9730 （0.9602-0.9817）） interval settings determined by the two optimization methods.The average time required by echocardiographic optimization and by QuickOptTM were （78.32±32.40） minutes and （1.98±1.64） minutes respectively （P 〈0.0001）.Conclusion The QuickOptTM algorithm provides a quicker,simpler and reliable alternative to the standard method for timing cycle optimization.（ClinicaITrial.gov Reference Number：NCT00918294）
文摘Two new ent-kaurene diterpenoids, angustifolin B and natural hydrogen-bonded 1:1 complex, angustifolin C, as well as four known diterpenoids, angustifolin, isodonal, sodoponin, tri-chorabdal B, were isolated from the leaves of Rabdosia angustifolia.
文摘Background Idiopathic hyperoxaluria （IH） may be caused by increased endogenous formation or exogenous absorption of oxalic acid.Characterization of the molecular pathogenesis of IH has been hampered by the lack of an ideal animal model.We therefore established a stabile rat IH model in order to analyze variation in gene expression profile in the jejunum and to investigate the association between IH pathogenesis and exogenous absorption of oxalic acid.Methods A rat model of IH was established and three female rats with IH were assigned to the study group,while three normal rats served as controls.Total RNA was isolated from the jejunum of rats in the two groups and mRNA was purified,reversely transcribed,labeled with Cy5 or Cy3 and hybridized to 27K Rat Genome Array.Differences in gene expression profile between the 2 groups were analyzed by bioinformatics methods.Results Comparative analysis revealed that the expression of 517 genes was up-regulated and that of 203 genes was down-regulated by at least two-fold in the jejunum of rats with idiopathic hyperoxaluria.These genes are related to many functions including cell signal transduction,DNA binding and transcription,ATP binding,ion binding and transport,cell receptors,immunity,cyclins,cytoskeleton structure,and metabolic proteins.Kyoto encyclopedia of genes and genomes （KEGG） signaling pathway analysis revealed that the variations of 239 pathway functional changes are statistically significant （P 〈0.05）.Conclusions cDNA microarray can be used effectively to screen differentially expressed genes in the jejunum of rats with idiopathic hyperoxaluria. These differentially expressed genes may underlie idiopathic hyperoxaluria pathophysiology and provide a platform for further studying molecular pathogenetic mechanisms.
