Our previous studies have successfully grafted biotin and galactose onto chitosan(CS)and synthesized biotin modified galactosylated chitosan(Bio-GC).The optimum N/P ratio of Bio-GC and plasmid DNA was 3:1.At this N/P ...Our previous studies have successfully grafted biotin and galactose onto chitosan(CS)and synthesized biotin modified galactosylated chitosan(Bio-GC).The optimum N/P ratio of Bio-GC and plasmid DNA was 3:1.At this N/P ratio,the transfection efficiency in the hepatoma cells was the highest with a slow release effect.Bio-GC nanomaterials exhibit the protective effect of preventing the gene from nuclease degradation,and can target the transfection into hepatoma cells by combination with galactose and biotin receptors.The transfection rate was inhibited by the competition of galactose and biotin.Bio-GC nanomaterials were imported into cells’cytoplasm by their receptors,followed by the imported exogenous gene transfected into the cells.Bio-GC nanomaterials can also cause inhibitory activity in the hepatoma cells in the model of orthotopic liver transplantation in mice,by carrying the gene through the blood to the hepatoma tissue.Taken together,bio-GC nanomaterials act as gene vectors with the activity of protecting the gene from DNase degradation,improving the rate of transfection in hepatoma cells,and transporting the gene into the cytoplasm in vitro and in vivo.Therefore,they are efficient hepatoma-targeting gene carriers.展开更多
This article reports on a retrospective analysis on 121 patients and a prospectivestudy on 21 patients with acute cholangitis of severe type(ACST)for a study on the timing se-lection of emergency operation for ACST.Tw...This article reports on a retrospective analysis on 121 patients and a prospectivestudy on 21 patients with acute cholangitis of severe type(ACST)for a study on the timing se-lection of emergency operation for ACST.Twenty two clinical,biological,etiologic,pathologicand operative variables were analyzed.Simple regression revealed 11 factors with prognosticsignificance,but multivariate analysis detected only 6 factors with independent significance inpredicting mortality(age,mean blood pressure,generalized peritonitis,serum albumin-globin ra-tio,blood culture,and the number of failed organs and systems).The results indicate that theclinical principles of treatment for ACST should be the combination of medical and surgicaltreatment.Active conservative treatment is practically applicable to the majority of ACST,espe-cially,those with short history and few complication.Prognostic mathematical model of ACSTdoes good for its timing selection of emergency operation.A critical level of 0.40 is determinedto be the discriminant score for emergency bile duct drainage.The model seems to have advan-tages over the traditional method.展开更多
Objective:To investigate glucose metabolic alterations in cerebral cortical subareas using ^(18)F-labeled glucose derivative fluorodeoxyglucose(FDG)micro-positron emission tomography(PET)scanning in a rat renal ischem...Objective:To investigate glucose metabolic alterations in cerebral cortical subareas using ^(18)F-labeled glucose derivative fluorodeoxyglucose(FDG)micro-positron emission tomography(PET)scanning in a rat renal ischemia/reperfusion(RIR)model.Methods:Small-animal PET imaging in vivo was performed with ^(18)F-labeled FDG as a PET tracer to identify glucose metabolic alterations in cerebral cortical subregions using a rat model of RIR.Results:We found that the average standardized uptake value(SUV_(average))of the cerebral cortical subareas in the RIR group was significantly increased compared to the sham group(P<0.05).We also found that glucose uptake in different cortical subregions including the left auditory cortex,right medial prefrontal cortex,right para cortex,left retrosplenial cortex,right retrosplenial cortex,and right visual cortex was significantly increased in the RIR group(P<0.05),but there was no significant difference in the SUV_(avcrage) of right auditory cortex,left medial prefrontal cortex,left para cortex,and left visual cortex between the two groups.Conclusion:The ^(18)F-FDG PET data suggests that RIR causes a profound shift in the metabolic machinery of cerebral cortex subregions.展开更多
Summary:Mechanisms of pruritus are implicated in the dysregulation of the metabolites in the spinal cord.We investigated pruritus behavioral testing in three groups of young adult male C57B1/6 mice,including one group...Summary:Mechanisms of pruritus are implicated in the dysregulation of the metabolites in the spinal cord.We investigated pruritus behavioral testing in three groups of young adult male C57B1/6 mice,including one group treated with normal saline,while the other groups intradermally injected with a-Me-5-HT(histamine-independent pruritogen),compound 48/80(histaminedependent pruritogen)at the nape skin of the neck,respectively.Proton nuclear magnetic resonance spectroscopy(MRS)was used to compare spinal metabolites from the vertebral cervical among three groups,and to study the association of spinal metabolite ratio and pruritus intensity.The MRS-measured N-acetylaspartate-to-myoinositol ratio(NAA/Ins)was significantly correlated with the number of scratches between normal saline group and 48/80 group or a-Me-5-HT group(both P<0.0001),indicating that NAA/Ins may be a robust surrogate marker of histamine-independent/dependent pruritogen.There was significant difference in Glu/Ins between normal saline group and 48/80 group(P=0.017),indicating that Glu/Ins may be a surrogate marker of histamine-dependent pruritogen,while GABA/Ins was highly significantly different between normal saline group and a-Me-5-HT group(P=0.008),suggesting that GABA/Ins may be a surrogate marker of histamineindependent pruritogen.MRS may reflect the extent of pruritus intensity elicited by a-Me-5-HT and compound 48/80 with sensitivity similar to the number of scratches,and above potential markers need to be further validated in pre-clinical and clinical treatment trials.展开更多
AIM: To investigate the efficacy of fu-qi granule(FQG) on carbon tetrachloride(CCl4) induced liver fibrosis in rats and the underlying mechanisms. METHODS: Sixty rats were randomly divided into sixgroups: normal contr...AIM: To investigate the efficacy of fu-qi granule(FQG) on carbon tetrachloride(CCl4) induced liver fibrosis in rats and the underlying mechanisms. METHODS: Sixty rats were randomly divided into sixgroups: normal control group, CCl4 induced liver fibrosis group, Anluo Huaxian Wan group and three treatment groups of FQG. Treatment of rats with intraperitoneal injection of carbon tetrachloride solution at 0.3 mL per 100 g body weigh twice a week for 8 wk. The normal control group the rats were given the media(olive oil) at the same time. In the first 2 wk, rats were raised with feedstuff(80% corn meal, 20% lard, 0.5% cholesterol). Serum samples were collected for alanine transaminase, aspartate aminotransferase, alkaline phosphatase, albumin, total protein assay and typical histopathological changes was observed in Hematoxylin-eosin staining sections. Smooth muscle alpha actin(α-SMA) was analyzed with immunohistochemistry. Mammalian target of rapamycin(m TOR) and hypoxia-inducible factor-1(HIF-1α) expressions were detected by Western blotting. Tissue inhibitor of matrix metalloproteinases-1(TIMP-1) and matrix metalloproteinases-9(MMP-9) were measured with semi-quantitative reverse transcriptasepolymerase chain reaction. RESULTS: FQG significantly reduced the serum levels of alanine transaminase, aspartate aminotransferase, alkaline phosphatase and increased the serum contents of albumin, total protein in rats with liver fibrosis. Moreover, FQG promoted extracellular matrix degradation by increasing MMP-9 and inhibiting TIMP-1 and α-SMA. m TOR and HIF-1α expression in liver significantly decreased in the rats treated with FQG. CONCLUSION: The results indicated that FQG significantly reverse fibrosis induced by CCl4, which should be developed as a new and promising preparation for the prevention of liver fibrosis.展开更多
Background Transforming growth factor-β1 (TGF-β1) exerts strong fibrogenic potential in culture-activated HSCs Smad 4 is a key intracellular mediator for the transforming growth factor-β (TGF-β) superfamily of gro...Background Transforming growth factor-β1 (TGF-β1) exerts strong fibrogenic potential in culture-activated HSCs Smad 4 is a key intracellular mediator for the transforming growth factor-β (TGF-β) superfamily of growth factors The aim of this study was to assess the effects of the antisense Smad 4 gene on Ito cell line, LI90 Methods The recombinant retroviral vector pLXSN-Smad 4 was constructed by cloning the rat antisense Smad 4 cDNA into the retroviral vector pLXSN Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad 4 and pLXSN vectors into PA317 cells Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418 The expression of Smad 4 was detected by Northern and Western blots Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed Results mRNA and protein expressions of Smad 4 in LI90 cells transfected with retrovirus containing the antisense Smad 4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells Cells hypoexpressing the Smad 4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline ( P <0 01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus Conclusions The antisense Smad 4 gene can suppress the expression of the Smad 4 gene, reduce endogenous production of Smad 4 mRNA and protein, block TGF-β1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM) Our results may provide a basis for the development of antifibrotic gene展开更多
Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely u...Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely used herbal medicine,has shown to cause idiosyncratic liver injury,but the underlying mechanisms are poorly understood.Increasing evidence has indicated that most cases of IDILI are immune mediated.Here,we report that icarisideⅡ(ICSⅡ),the major active and metabolic constituent of EF,causes idiosyncratic liver injury by promoting NLRP3 inflammasome activation.ICSⅡexacerbates NLRP3 inflammasome activation triggered by adenosine triphosphate(ATP)and nigericin,but not silicon dioxide(SiO2),monosodium urate(MSU)crystal or cytosolic lipopolysaccharide(LPS).Additionally,the activation of NLRC4 and AIM2 inflammasomes is not affected by ICSⅡ.Mechanistically,synergistic induction of mitochondrial reactive oxygen species(mtROS)is a crucial contributor to the enhancing effect of ICSⅡon ATP-or nigericin-induced NLRP3 inflammasome activation.Importantly,in vivo data show that a combination of non-hepatotoxic doses of LPS and ICSⅡcauses the increase of aminotransferase activity,hepatic inflammation and pyroptosis,which is attenuated by Nlrp3 deficiency or pretreatment with MCC950(a specific NLRP3 inflammasome inhibitor).In conclusion,these findings demonstrate that ICSⅡcauses idiosyncratic liver injury through enhancing NLRP3 inflammasome activation and suggest that ICSⅡmay be a risk factor and responsible for EF-induced liver injury.展开更多
基金Funded by the Scientific Research Project of Shanghai Municipal Health Commission(No.201940430)。
文摘Our previous studies have successfully grafted biotin and galactose onto chitosan(CS)and synthesized biotin modified galactosylated chitosan(Bio-GC).The optimum N/P ratio of Bio-GC and plasmid DNA was 3:1.At this N/P ratio,the transfection efficiency in the hepatoma cells was the highest with a slow release effect.Bio-GC nanomaterials exhibit the protective effect of preventing the gene from nuclease degradation,and can target the transfection into hepatoma cells by combination with galactose and biotin receptors.The transfection rate was inhibited by the competition of galactose and biotin.Bio-GC nanomaterials were imported into cells’cytoplasm by their receptors,followed by the imported exogenous gene transfected into the cells.Bio-GC nanomaterials can also cause inhibitory activity in the hepatoma cells in the model of orthotopic liver transplantation in mice,by carrying the gene through the blood to the hepatoma tissue.Taken together,bio-GC nanomaterials act as gene vectors with the activity of protecting the gene from DNase degradation,improving the rate of transfection in hepatoma cells,and transporting the gene into the cytoplasm in vitro and in vivo.Therefore,they are efficient hepatoma-targeting gene carriers.
基金Supported by the National Natural Science Foundation of China
文摘This article reports on a retrospective analysis on 121 patients and a prospectivestudy on 21 patients with acute cholangitis of severe type(ACST)for a study on the timing se-lection of emergency operation for ACST.Twenty two clinical,biological,etiologic,pathologicand operative variables were analyzed.Simple regression revealed 11 factors with prognosticsignificance,but multivariate analysis detected only 6 factors with independent significance inpredicting mortality(age,mean blood pressure,generalized peritonitis,serum albumin-globin ra-tio,blood culture,and the number of failed organs and systems).The results indicate that theclinical principles of treatment for ACST should be the combination of medical and surgicaltreatment.Active conservative treatment is practically applicable to the majority of ACST,espe-cially,those with short history and few complication.Prognostic mathematical model of ACSTdoes good for its timing selection of emergency operation.A critical level of 0.40 is determinedto be the discriminant score for emergency bile duct drainage.The model seems to have advan-tages over the traditional method.
基金supported by grants from Hubei Provincial Natural Science Foundation of China(No.2019CFB805)National Natural Science Foundation of China(No.81670240 and No.81873467).
文摘Objective:To investigate glucose metabolic alterations in cerebral cortical subareas using ^(18)F-labeled glucose derivative fluorodeoxyglucose(FDG)micro-positron emission tomography(PET)scanning in a rat renal ischemia/reperfusion(RIR)model.Methods:Small-animal PET imaging in vivo was performed with ^(18)F-labeled FDG as a PET tracer to identify glucose metabolic alterations in cerebral cortical subregions using a rat model of RIR.Results:We found that the average standardized uptake value(SUV_(average))of the cerebral cortical subareas in the RIR group was significantly increased compared to the sham group(P<0.05).We also found that glucose uptake in different cortical subregions including the left auditory cortex,right medial prefrontal cortex,right para cortex,left retrosplenial cortex,right retrosplenial cortex,and right visual cortex was significantly increased in the RIR group(P<0.05),but there was no significant difference in the SUV_(avcrage) of right auditory cortex,left medial prefrontal cortex,left para cortex,and left visual cortex between the two groups.Conclusion:The ^(18)F-FDG PET data suggests that RIR causes a profound shift in the metabolic machinery of cerebral cortex subregions.
基金This work was supported by grants from the National Natural Science Foundation of China(No.81670240 and No.81873467)the Medical Innovation Project in Fujian Province(No.2017-CX-48).
文摘Summary:Mechanisms of pruritus are implicated in the dysregulation of the metabolites in the spinal cord.We investigated pruritus behavioral testing in three groups of young adult male C57B1/6 mice,including one group treated with normal saline,while the other groups intradermally injected with a-Me-5-HT(histamine-independent pruritogen),compound 48/80(histaminedependent pruritogen)at the nape skin of the neck,respectively.Proton nuclear magnetic resonance spectroscopy(MRS)was used to compare spinal metabolites from the vertebral cervical among three groups,and to study the association of spinal metabolite ratio and pruritus intensity.The MRS-measured N-acetylaspartate-to-myoinositol ratio(NAA/Ins)was significantly correlated with the number of scratches between normal saline group and 48/80 group or a-Me-5-HT group(both P<0.0001),indicating that NAA/Ins may be a robust surrogate marker of histamine-independent/dependent pruritogen.There was significant difference in Glu/Ins between normal saline group and 48/80 group(P=0.017),indicating that Glu/Ins may be a surrogate marker of histamine-dependent pruritogen,while GABA/Ins was highly significantly different between normal saline group and a-Me-5-HT group(P=0.008),suggesting that GABA/Ins may be a surrogate marker of histamineindependent pruritogen.MRS may reflect the extent of pruritus intensity elicited by a-Me-5-HT and compound 48/80 with sensitivity similar to the number of scratches,and above potential markers need to be further validated in pre-clinical and clinical treatment trials.
基金Supported by The National Natural Sciences Foundation,No.81173571National Basic Research Program of China,No.2007CB512607The Major Projects of the National Science and Technology,No.2012ZX10005010-002-002
文摘AIM: To investigate the efficacy of fu-qi granule(FQG) on carbon tetrachloride(CCl4) induced liver fibrosis in rats and the underlying mechanisms. METHODS: Sixty rats were randomly divided into sixgroups: normal control group, CCl4 induced liver fibrosis group, Anluo Huaxian Wan group and three treatment groups of FQG. Treatment of rats with intraperitoneal injection of carbon tetrachloride solution at 0.3 mL per 100 g body weigh twice a week for 8 wk. The normal control group the rats were given the media(olive oil) at the same time. In the first 2 wk, rats were raised with feedstuff(80% corn meal, 20% lard, 0.5% cholesterol). Serum samples were collected for alanine transaminase, aspartate aminotransferase, alkaline phosphatase, albumin, total protein assay and typical histopathological changes was observed in Hematoxylin-eosin staining sections. Smooth muscle alpha actin(α-SMA) was analyzed with immunohistochemistry. Mammalian target of rapamycin(m TOR) and hypoxia-inducible factor-1(HIF-1α) expressions were detected by Western blotting. Tissue inhibitor of matrix metalloproteinases-1(TIMP-1) and matrix metalloproteinases-9(MMP-9) were measured with semi-quantitative reverse transcriptasepolymerase chain reaction. RESULTS: FQG significantly reduced the serum levels of alanine transaminase, aspartate aminotransferase, alkaline phosphatase and increased the serum contents of albumin, total protein in rats with liver fibrosis. Moreover, FQG promoted extracellular matrix degradation by increasing MMP-9 and inhibiting TIMP-1 and α-SMA. m TOR and HIF-1α expression in liver significantly decreased in the rats treated with FQG. CONCLUSION: The results indicated that FQG significantly reverse fibrosis induced by CCl4, which should be developed as a new and promising preparation for the prevention of liver fibrosis.
文摘Background Transforming growth factor-β1 (TGF-β1) exerts strong fibrogenic potential in culture-activated HSCs Smad 4 is a key intracellular mediator for the transforming growth factor-β (TGF-β) superfamily of growth factors The aim of this study was to assess the effects of the antisense Smad 4 gene on Ito cell line, LI90 Methods The recombinant retroviral vector pLXSN-Smad 4 was constructed by cloning the rat antisense Smad 4 cDNA into the retroviral vector pLXSN Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad 4 and pLXSN vectors into PA317 cells Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418 The expression of Smad 4 was detected by Northern and Western blots Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed Results mRNA and protein expressions of Smad 4 in LI90 cells transfected with retrovirus containing the antisense Smad 4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells Cells hypoexpressing the Smad 4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline ( P <0 01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus Conclusions The antisense Smad 4 gene can suppress the expression of the Smad 4 gene, reduce endogenous production of Smad 4 mRNA and protein, block TGF-β1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM) Our results may provide a basis for the development of antifibrotic gene
基金supported by National Natural Science Foundation of China(81874368,81630100,and 81903891)Beijing Nova Program(Z181100006218001,China)+1 种基金National Science&Technology Major Project“Key New Drug Creation and Manufacturing Program”(2017ZX09301022 and 2018ZX09101002-001-002,China)the Innovation Groups of the National Natural Science Foundation of China(81721002)
文摘Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely used herbal medicine,has shown to cause idiosyncratic liver injury,but the underlying mechanisms are poorly understood.Increasing evidence has indicated that most cases of IDILI are immune mediated.Here,we report that icarisideⅡ(ICSⅡ),the major active and metabolic constituent of EF,causes idiosyncratic liver injury by promoting NLRP3 inflammasome activation.ICSⅡexacerbates NLRP3 inflammasome activation triggered by adenosine triphosphate(ATP)and nigericin,but not silicon dioxide(SiO2),monosodium urate(MSU)crystal or cytosolic lipopolysaccharide(LPS).Additionally,the activation of NLRC4 and AIM2 inflammasomes is not affected by ICSⅡ.Mechanistically,synergistic induction of mitochondrial reactive oxygen species(mtROS)is a crucial contributor to the enhancing effect of ICSⅡon ATP-or nigericin-induced NLRP3 inflammasome activation.Importantly,in vivo data show that a combination of non-hepatotoxic doses of LPS and ICSⅡcauses the increase of aminotransferase activity,hepatic inflammation and pyroptosis,which is attenuated by Nlrp3 deficiency or pretreatment with MCC950(a specific NLRP3 inflammasome inhibitor).In conclusion,these findings demonstrate that ICSⅡcauses idiosyncratic liver injury through enhancing NLRP3 inflammasome activation and suggest that ICSⅡmay be a risk factor and responsible for EF-induced liver injury.
