Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the pr...Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the prodrug of epigallocatechin gallate(ProEGCG)exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate(EGCG).However,their direct binding targets and underlying mechanisms for the differential effects remain unknown.In this study,we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis.Additionally,1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin(MTDH)and PX domain containing serine/threonine kinase-like(PXK)as novel binding targets of EGCG and ProEGCG,respectively.Computational simulation and BioLayer interferometry were used to confirm their binding affinity.Our results showed that MTDH-EGCG inhibited protein kinase B(Akt)-mediated angiogenesis,while PXK-ProEGCG inhibited epidermal growth factor(EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor(HIF-1a)/vascular endothelial growth factor(VEGF)pathway.In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways.Moreover,our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.展开更多
Chronic denervation is one of the key factors that affect nerve regeneration.Chronic axotomy deteriorates the distal nerve stump,causes protein changes,and renders the microenvironment less permissive for regeneration...Chronic denervation is one of the key factors that affect nerve regeneration.Chronic axotomy deteriorates the distal nerve stump,causes protein changes,and renders the microenvironment less permissive for regeneration.Some of these factors/proteins have been individually studied.To better delineate the comprehensive protein expression profiles and identify proteins that contribute to or are associated with this detrimental effect,we carried out a proteomic analysis of the distal nerve using an established delayed rat sciatic nerve repair model.Four rats that received immediate repair after sciatic nerve transection served as control,whereas four rats in the experimental group(chronic denervation)had their sciatic nerve repaired after a 12-week delay.All the rats were sacrificed after 16 weeks to harvest the distal nerves for extracting proteins.Twenty-five micrograms of protein from each sample were fractionated in SDS-PAGE gels.NanoLC-MS/MS analysis was applied to the gels.Protein expression levels of nerves on the surgery side were compared to those on the contralateral side.Any protein with a P value of less than 0.05 and a fold change of 4 or higher was deemed differentially expressed.All the differentially expressed proteins in both groups were further stratified according to the biological processes.A PubMed search was also conducted to identify the differentially expressed proteins that have been reported to be either beneficial or detrimental to nerve regeneration.Ingenuity Pathway Analysis(IPA)software was used for pathway analysis.The results showed that 709 differentially expressed proteins were identified in the delayed repair group,with a bigger proportion of immune and inflammatory process-related proteins and a smaller proportion of proteins related to axon regeneration and lipid metabolism in comparison to the control group where 478 differentially expressed proteins were identified.The experimental group also had more beneficial proteins that were downregulated and more detrimental proteins that were upregulated.IPA revealed that protective pathways such as LXR/RXR,acute phase response,RAC,ERK/MAPK,CNTF,IL-6,and FGF signaling were inhibited in the delayed repair group,whereas three detrimental pathways,including the complement system,PTEN,and apoptosis signaling,were activated.An available database of the adult rodent sciatic nerve was used to assign protein changes to specific cell types.The poor regeneration seen in the delayed repair group could be associated with the down-regulation of beneficial proteins and up-regulation of detrimental proteins.The proteins and pathways identified in this study may offer clues for future studies to identify therapeutic targets.展开更多
Aims： High-quality DNA as input material for a NGS （next-generation sequencing） workflow is essential for the successful preparation of a DNA library. Additionally, DNA quality has a strong impact on sequencing res...Aims： High-quality DNA as input material for a NGS （next-generation sequencing） workflow is essential for the successful preparation of a DNA library. Additionally, DNA quality has a strong impact on sequencing results. Therefore, it is important to include QC （quality control） steps to assess size, concentration, molarity, and integrity of the DNA during the workflow. Material and Methods： The WES （Whole Exome Sequencing） workflow at the Genomics and Proteomics Core Facility of DKFZ （the German Cancer Research Center） was performed with several QC steps： QC of the input material, QC of intermediate products during library preparation, and QC of final libraries. The Agilent 4200 TapeStation system, which offers automated sample processing, was used to evaluate quantity, size, molarity, and integrity of the samples. Key Findings： The Agilent Genomic DNA ScreenTape assay offers an unbiased genomic DNA integrity assessment, which enables protocol adaption for optimized library preparation, for example, selection of a suitable shearing protocol. Additionally, QC steps during library preparation such as evaluation of library size, concentration, and molarity ensure maximal sequencing output. Significance： The automated and fast high-throughput analysis of genomic DNA with the 4200 TapeStation system helps to save labor time and costs. Additionally, the easy-to-use system can be integrated as a QC tool into the NGS workflow to ensure successful library preparation. QC steps enable the confirmation of suitable library size and concentration for the workflow.展开更多
Background:Determining the etiology of biliary strictures is challenging,and the sensitivities of the current tests to diagnose them are low.Protein biomarkers in bile,in combination with other tests,may improve sensi...Background:Determining the etiology of biliary strictures is challenging,and the sensitivities of the current tests to diagnose them are low.Protein biomarkers in bile,in combination with other tests,may improve sensitivity in diagnosing biliary strictures.Objective:To analyse the differential abundance of proteins in benign and malignant biliary strictures through proteomic analysis of bile.Methods:In this prospective,cross-sectional study,bile was aspirated in 24 patients undergoing endoscopic retrograde cholangiopancreatography(ERCP)including six patients with primary sclerosing cholangitis(PSC),three with cholangiocarcinoma(CCA),ten with pancreatic cancer,and five with benign biliary conditions.Liquid chromatography/mass spectrometry was used to examine the bile for differential abundance of protein biomarkers.The relative abundance of various proteins was compared in the malignant vs.benign groups and in CCA vs.PSC.Results:The majority of the proteins identified in bile were similar to those of the plasma(plasma proteins)and certain proteins were differentially expressed among the different groups(CCA,pancreatic cancer,PSC or benign).A total of 18 proteins were identified as being more abundant in the malignant group(CCA and pancreatic cancer)than in the benign strictures group,including myeloperoxidase,complement C3,inter-alpha-trypsin inhibitor heavy chain H4,apolipoprotein B-100,and kininogen-1 isoform 2.A total of 30 proteins were identified to be less abundant in the malignant group than in the benign group,including trefoil factor 2,superoxide dismutase[Cu-Zn],kallikrein-1,carboxypeptidase B and trefoil factor 1.Conclusions:Protein biomarkers in bile may differentiate malignant from benign biliary strictures.Larger studies are warranted to validate these observations.展开更多
Magnesium alloys containing biocompatible components show tremendous promise for applications as temporary biomedical devices. However, to ensure their safe use as biodegradeable implants, it is essential to control t...Magnesium alloys containing biocompatible components show tremendous promise for applications as temporary biomedical devices. However, to ensure their safe use as biodegradeable implants, it is essential to control their corrosion rates. In concentrated Mg alloys, a microgalvanic coupling between the α-Mg matrix and secondary precipitates exists which results in increased corrosion rate. To address this challenge, we engineered the microstructure of a biodegradable Mg-Zn-RE-Zr alloy by friction stir processing (FSP), improving its corrosion resistance and mechanical properties simultaneously. The FS processed alloy with refined grains and broken and uniformly distributed secondary precipitates showed a relatively uniform corrosion morphology accompanied with the formation of a stable passive layer on the alloy surface. In vivo corrosion evaluation of the processed alloy in a small animal model showed that the material was well-tolerated with no signs of inflammation or harmful by-products. Remarkably, the processed alloy supported bone until it healed till eight weeks with a low in vivo corrosion rate of 0.7 mm/year. Moreover, we analyzed blood and histology of the critical organs such as liver and kidney, which showed normal functionality and consistent ion and enzyme levels, throughout the 12- week study period. These results demonstrate that the processed Mg-Zn-RE-Zr alloy offers promising potential for osseointegration in bone tissue healing while also exhibiting controlled biodegradability due to its engineered microstructure. The results from the present study will have profound benefit for bone fracture management, particularly in pediatric and elderly patients.展开更多
基金supported by the GRF RGC&CRF,Hong Kong(Grant Nos.:475012 and C5045-20 EF)HMRF,Hong Kong(Grant No.:03141386)+3 种基金ITF,Hong Kong(Grant No.:ITS/209/12)UGC Direct Grant 2011,2012,2021.032HKOG Trust Fund 2011,2014,2019the National Natural Science Foundation of China(Grant Nos.:81974225 and 82201823)。
文摘Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus.Angiogenesis is a major pathophysiology in endometriosis.Our previous studies have demonstrated that the prodrug of epigallocatechin gallate(ProEGCG)exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate(EGCG).However,their direct binding targets and underlying mechanisms for the differential effects remain unknown.In this study,we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis.Additionally,1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin(MTDH)and PX domain containing serine/threonine kinase-like(PXK)as novel binding targets of EGCG and ProEGCG,respectively.Computational simulation and BioLayer interferometry were used to confirm their binding affinity.Our results showed that MTDH-EGCG inhibited protein kinase B(Akt)-mediated angiogenesis,while PXK-ProEGCG inhibited epidermal growth factor(EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor(HIF-1a)/vascular endothelial growth factor(VEGF)pathway.In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways.Moreover,our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.
基金supported by Helene Houle Career Development Award in Neurologic Surgery Research and Fund for Mayo Clinic Center for Regenerative Medicine Program Director,Neuroregenerative Medicine,Mayo Clinic College of Medicine and Science.SG was supported by the Chinese Scholarship Council.
文摘Chronic denervation is one of the key factors that affect nerve regeneration.Chronic axotomy deteriorates the distal nerve stump,causes protein changes,and renders the microenvironment less permissive for regeneration.Some of these factors/proteins have been individually studied.To better delineate the comprehensive protein expression profiles and identify proteins that contribute to or are associated with this detrimental effect,we carried out a proteomic analysis of the distal nerve using an established delayed rat sciatic nerve repair model.Four rats that received immediate repair after sciatic nerve transection served as control,whereas four rats in the experimental group(chronic denervation)had their sciatic nerve repaired after a 12-week delay.All the rats were sacrificed after 16 weeks to harvest the distal nerves for extracting proteins.Twenty-five micrograms of protein from each sample were fractionated in SDS-PAGE gels.NanoLC-MS/MS analysis was applied to the gels.Protein expression levels of nerves on the surgery side were compared to those on the contralateral side.Any protein with a P value of less than 0.05 and a fold change of 4 or higher was deemed differentially expressed.All the differentially expressed proteins in both groups were further stratified according to the biological processes.A PubMed search was also conducted to identify the differentially expressed proteins that have been reported to be either beneficial or detrimental to nerve regeneration.Ingenuity Pathway Analysis(IPA)software was used for pathway analysis.The results showed that 709 differentially expressed proteins were identified in the delayed repair group,with a bigger proportion of immune and inflammatory process-related proteins and a smaller proportion of proteins related to axon regeneration and lipid metabolism in comparison to the control group where 478 differentially expressed proteins were identified.The experimental group also had more beneficial proteins that were downregulated and more detrimental proteins that were upregulated.IPA revealed that protective pathways such as LXR/RXR,acute phase response,RAC,ERK/MAPK,CNTF,IL-6,and FGF signaling were inhibited in the delayed repair group,whereas three detrimental pathways,including the complement system,PTEN,and apoptosis signaling,were activated.An available database of the adult rodent sciatic nerve was used to assign protein changes to specific cell types.The poor regeneration seen in the delayed repair group could be associated with the down-regulation of beneficial proteins and up-regulation of detrimental proteins.The proteins and pathways identified in this study may offer clues for future studies to identify therapeutic targets.
文摘Aims： High-quality DNA as input material for a NGS （next-generation sequencing） workflow is essential for the successful preparation of a DNA library. Additionally, DNA quality has a strong impact on sequencing results. Therefore, it is important to include QC （quality control） steps to assess size, concentration, molarity, and integrity of the DNA during the workflow. Material and Methods： The WES （Whole Exome Sequencing） workflow at the Genomics and Proteomics Core Facility of DKFZ （the German Cancer Research Center） was performed with several QC steps： QC of the input material, QC of intermediate products during library preparation, and QC of final libraries. The Agilent 4200 TapeStation system, which offers automated sample processing, was used to evaluate quantity, size, molarity, and integrity of the samples. Key Findings： The Agilent Genomic DNA ScreenTape assay offers an unbiased genomic DNA integrity assessment, which enables protocol adaption for optimized library preparation, for example, selection of a suitable shearing protocol. Additionally, QC steps during library preparation such as evaluation of library size, concentration, and molarity ensure maximal sequencing output. Significance： The automated and fast high-throughput analysis of genomic DNA with the 4200 TapeStation system helps to save labor time and costs. Additionally, the easy-to-use system can be integrated as a QC tool into the NGS workflow to ensure successful library preparation. QC steps enable the confirmation of suitable library size and concentration for the workflow.
基金supported by a research grant from the American College of Gastroenterology(ACG)(to U.N)in part by the National Institutes of Health,National Center for Research Resources,CTSA,UL1TR 000439-06 Cleveland,OhioObitrap Elite Instrument was purchased from an NIH S10 shared instrument grant(1S10RR031537-01).
文摘Background:Determining the etiology of biliary strictures is challenging,and the sensitivities of the current tests to diagnose them are low.Protein biomarkers in bile,in combination with other tests,may improve sensitivity in diagnosing biliary strictures.Objective:To analyse the differential abundance of proteins in benign and malignant biliary strictures through proteomic analysis of bile.Methods:In this prospective,cross-sectional study,bile was aspirated in 24 patients undergoing endoscopic retrograde cholangiopancreatography(ERCP)including six patients with primary sclerosing cholangitis(PSC),three with cholangiocarcinoma(CCA),ten with pancreatic cancer,and five with benign biliary conditions.Liquid chromatography/mass spectrometry was used to examine the bile for differential abundance of protein biomarkers.The relative abundance of various proteins was compared in the malignant vs.benign groups and in CCA vs.PSC.Results:The majority of the proteins identified in bile were similar to those of the plasma(plasma proteins)and certain proteins were differentially expressed among the different groups(CCA,pancreatic cancer,PSC or benign).A total of 18 proteins were identified as being more abundant in the malignant group(CCA and pancreatic cancer)than in the benign strictures group,including myeloperoxidase,complement C3,inter-alpha-trypsin inhibitor heavy chain H4,apolipoprotein B-100,and kininogen-1 isoform 2.A total of 30 proteins were identified to be less abundant in the malignant group than in the benign group,including trefoil factor 2,superoxide dismutase[Cu-Zn],kallikrein-1,carboxypeptidase B and trefoil factor 1.Conclusions:Protein biomarkers in bile may differentiate malignant from benign biliary strictures.Larger studies are warranted to validate these observations.
文摘Magnesium alloys containing biocompatible components show tremendous promise for applications as temporary biomedical devices. However, to ensure their safe use as biodegradeable implants, it is essential to control their corrosion rates. In concentrated Mg alloys, a microgalvanic coupling between the α-Mg matrix and secondary precipitates exists which results in increased corrosion rate. To address this challenge, we engineered the microstructure of a biodegradable Mg-Zn-RE-Zr alloy by friction stir processing (FSP), improving its corrosion resistance and mechanical properties simultaneously. The FS processed alloy with refined grains and broken and uniformly distributed secondary precipitates showed a relatively uniform corrosion morphology accompanied with the formation of a stable passive layer on the alloy surface. In vivo corrosion evaluation of the processed alloy in a small animal model showed that the material was well-tolerated with no signs of inflammation or harmful by-products. Remarkably, the processed alloy supported bone until it healed till eight weeks with a low in vivo corrosion rate of 0.7 mm/year. Moreover, we analyzed blood and histology of the critical organs such as liver and kidney, which showed normal functionality and consistent ion and enzyme levels, throughout the 12- week study period. These results demonstrate that the processed Mg-Zn-RE-Zr alloy offers promising potential for osseointegration in bone tissue healing while also exhibiting controlled biodegradability due to its engineered microstructure. The results from the present study will have profound benefit for bone fracture management, particularly in pediatric and elderly patients.
