AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in O...AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. ' RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group. CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.展开更多
In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(a...In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(and often naturally occurring)staining substances.Incidentally,the active ingredients of most of them were small molecules.With the advent of time,the knowledge of chemistry helped identify compounds and conditions for staining.The staining reagents were even found to enhance the visibility of the organelles.Silver impregnation identification of Golgi bodies was discovered in owl optic nerve.Staining reagents since the late 1800s were widely used across all disciplines and for nerve tissue and became a key contributor to advancement in nerve-related research.The use of these reagents provided insight into the organization of the neuronal tissues and helped distinguish nerve degeneration from regeneration.The neuronal staining reagents have played a fundamental role in the clinical research facilitating the identification of biological mechanisms underlying eye and neuropsychiatric diseases.We found a lack of systematic description of all staining reagents,whether they had been used historically or currently used.There is a lack of readily available information for optimal staining of different neuronal tissues for a given purpose.We present here a grouping of the reagents based on their target location:(I)the central nervous system(CNS),(II)the peripheral nervous system(PNS),or(III)both.The biochemical reactions of most of the staining reagents is based on acidic or basic pH and specific reaction partners such as organelle or biomolecules that exists within the given tissue type.We present here a summary of the chemical composition,optimal staining condition,use for given neuronal tissue and,where possible,historic usage.Several biomolecules such as lipids and metabolites lack specific antibodies.Despite being non-specific the reagents enhance contrast and provide corroboration about the microenvironment.In future,these reagents in combination with emerging techniques such as imaging mass spectrometry and kinetic histochemistry will validate or expand our understanding of localization of molecules within tissues or cells that are important for ophthalmology and vision science.展开更多
This study investigates the variability in cancer diagnosis across different tissues and organs, with a focus on the role of diagnostic methods such as Hematoxylin and Eosin (H&E) staining and immunohistochemistry...This study investigates the variability in cancer diagnosis across different tissues and organs, with a focus on the role of diagnostic methods such as Hematoxylin and Eosin (H&E) staining and immunohistochemistry (IHC). The predominance of female breast cancer (30%) aligns with global trends, underscoring the need for robust diagnostic protocols, particularly in developing regions. Other prevalent cancers, including skin, stomach, and cervix uteri, reflect a mix of environmental, genetic, and infectious factors. The underrepresentation of gallbladder and thyroid cancers (<1%) suggests potential underdiagnosis or lower prevalence. Age distribution data indicate peak cancer incidence in individuals aged 31 - 45 years, with gender-specific cancers like breast and cervical cancer predominantly affecting females (63.4%). The analysis also highlights significant diagnostic gaps, as 61.2% of cases did not undergo IHC testing due to resource constraints, leading to potential biases in cancer prevalence and diagnostic accuracy. The study emphasizes the complementary role of IHC in confirming ambiguous H&E findings, with strong alignment observed when both methods were used. However, the absence of IHC in many cases limits the robustness of conclusions, suggesting the need for increased access to IHC testing. The findings advocate for integrating IHC into routine diagnostics, expanding diagnostic capabilities, and improving sample sizes to ensure more reliable and comprehensive cancer data.展开更多
Electroacupuncture(EA)has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia.However,there are few studies on the results and mechanism of t...Electroacupuncture(EA)has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia.However,there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects.In this study,EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed.Except for the normal group,rats in other groups were fed a high-fat diet throughout the whole experiment.Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks.Middle cerebral artery thrombus models were induced by pasting 50%FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group.EA1 group rats received EA at bilateral ST40(Fenglong)for 7 days before the thrombosis.Rats in the EA1 and EA2 groups received EA at GV20(Baihui)and bilateral ST40 for 14 days after model establishment.Neuronal health was assessed by hematoxylin-eosin staining in the brain.Hyperlipidemia was assessed by biochemical methods that measured total cholesterol,triglyceride,low-density lipoprotein and high-density lipoprotein in blood sera.Behavioral analysis was used to confirm the establishment of the model.Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region.The results demonstrated that,compared with the model group,blood lipid levels significantly decreased,glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups.The repair effect was superior in the EA1 group than in the EA2 group.These findings confirm that EA can reduce blood lipid,inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis.All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine,China(approval No.BUCM-3-2018022802-1002)on April 12,2018.展开更多
BACKGROUND Diabetes is a metabolic disease with a high complication rate.Diabetic foot ulcers(DFUs)seriously affect the quality of life of patients.A total of 15%-20%of diabetic patients develop DFUs,which heal with d...BACKGROUND Diabetes is a metabolic disease with a high complication rate.Diabetic foot ulcers(DFUs)seriously affect the quality of life of patients.A total of 15%-20%of diabetic patients develop DFUs,which heal with difficulty over a long time and can result in amputation and disability.Traditional Chinese medicine has a unique effect in the treatment of skin ulcerative diseases.Ruyi Jinhuang powder(RHP)is one of the classic prescriptions in traditional Chinese medicine and is widely used in clinical practice.AIM To verify the ability of RHP to promote wound healing by electron microscopy analysis in animal models and hematoxylin-eosin(HE)staining.The effective components of RHP were extracted and identified by gas chromatography-mass spectrometry(GC-MS),and the obtained chemical components were analyzed by network pharmacology methods to predict its therapeutic mechanism.METHODS Sprague Dawley rats were injected with streptozotocin to establish the DFU model.HE staining was used to observe the wound tissue under an electron microscope.The chemical constituents of RHP were extracted first by supercritical fluid extraction and alcohol extraction,and then,GC-MS and ultra-performance liquid chromatography–MS were used to separately identify the chemical constituents.In addition,the"herb-component-target"link was established through the Traditional Chinese Medicine Systems Pharmacology database to obtain the target information,and the molecular docking of important components and key targets was performed in Discovery Studio software.Cytoscape software was used to visualize and analyze the relationship between the chemical composition,targets and Traditional Chinese Medicine network.RESULTS RHP promoted DFU healing in rats by affecting fibroblasts and nerve cells.A total of 89 chemical components were obtained by GC-MS.Network pharmacological analysis revealed that RHP was associated with 36 targets and 27 pathways in the treatment of DFU,of which the important components were luteolin,trans caryophyllene,ar-turmerone,palmitic acid,methyl palmitate,gallic acid,demethoxycurcumin,berberine,and rheic acid.The key targets were posttranscriptional silencing,topoisomerase II alpha,muscarinic acetylcholine receptor M2,interleukin 6,tumor necrosis factor and retinoic X receptor alpha,and the key pathways were the phosphoinositide 3-kinase-protein kinase B signaling pathway,neuroactive ligand–receptor interactions,and the forkhead box O signaling pathway.CONCLUSION Our results indicated that RHP may play a role in the treatment of DFU through these target pathways by affecting insulin resistance,altering the nervous system and immune system,participating in inflammatory responses and regulating cell proliferation,differentiation and apoptosis through other specific mechanisms.展开更多
According to clinical statistics,the mortality of patients with early brainstem hemorrhage is high.In this study,we established rat models of brainstem hemorrhage by injecting type Ⅶ collagenase into the right basote...According to clinical statistics,the mortality of patients with early brainstem hemorrhage is high.In this study,we established rat models of brainstem hemorrhage by injecting type Ⅶ collagenase into the right basotegmental pontine and investigated the pathological changes of early brainstem hemorrhage using multi-sequence magnetic resonance imaging and histopathological methods.We found that brainstem hematoma gradually formed in the injured rats over the first 3 days and then reduced after 7 days.The edema that occurred was mainly of the vasogenic type.No complete myelin sheath structure was found around the focus of the brainstem hemorrhage.The integrity and continuity of nerve fibers gradually deteriorated over the first 7 days.Neuronal degeneration was mild in the first 3 days and then obviously aggravated on the 7^(th)day.Inflammatory cytokines,interleukin-1β,and tumor necrosis factorαappeared on the 1st day after intracerebral hemorrhage,reached peak levels on the 3^(rd)day,and decreased from the 7^(th)day.Our findings show the characteristics of the progression of early brainstem hemorrhage.展开更多
基金Supported by National Natural Science Foundation of China(No.30973899)
文摘AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. ' RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group. CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.
基金supported by an unrestricted grant from Research to Prevent Blindness and NIH grants EY14801,EY031292.
文摘In the early days of deciphering the injured neuronal tissues led to the realization that contrast is necessary to discern the parts of the recovering tissues from the damaged ones.Early attempts relied on available(and often naturally occurring)staining substances.Incidentally,the active ingredients of most of them were small molecules.With the advent of time,the knowledge of chemistry helped identify compounds and conditions for staining.The staining reagents were even found to enhance the visibility of the organelles.Silver impregnation identification of Golgi bodies was discovered in owl optic nerve.Staining reagents since the late 1800s were widely used across all disciplines and for nerve tissue and became a key contributor to advancement in nerve-related research.The use of these reagents provided insight into the organization of the neuronal tissues and helped distinguish nerve degeneration from regeneration.The neuronal staining reagents have played a fundamental role in the clinical research facilitating the identification of biological mechanisms underlying eye and neuropsychiatric diseases.We found a lack of systematic description of all staining reagents,whether they had been used historically or currently used.There is a lack of readily available information for optimal staining of different neuronal tissues for a given purpose.We present here a grouping of the reagents based on their target location:(I)the central nervous system(CNS),(II)the peripheral nervous system(PNS),or(III)both.The biochemical reactions of most of the staining reagents is based on acidic or basic pH and specific reaction partners such as organelle or biomolecules that exists within the given tissue type.We present here a summary of the chemical composition,optimal staining condition,use for given neuronal tissue and,where possible,historic usage.Several biomolecules such as lipids and metabolites lack specific antibodies.Despite being non-specific the reagents enhance contrast and provide corroboration about the microenvironment.In future,these reagents in combination with emerging techniques such as imaging mass spectrometry and kinetic histochemistry will validate or expand our understanding of localization of molecules within tissues or cells that are important for ophthalmology and vision science.
文摘This study investigates the variability in cancer diagnosis across different tissues and organs, with a focus on the role of diagnostic methods such as Hematoxylin and Eosin (H&E) staining and immunohistochemistry (IHC). The predominance of female breast cancer (30%) aligns with global trends, underscoring the need for robust diagnostic protocols, particularly in developing regions. Other prevalent cancers, including skin, stomach, and cervix uteri, reflect a mix of environmental, genetic, and infectious factors. The underrepresentation of gallbladder and thyroid cancers (<1%) suggests potential underdiagnosis or lower prevalence. Age distribution data indicate peak cancer incidence in individuals aged 31 - 45 years, with gender-specific cancers like breast and cervical cancer predominantly affecting females (63.4%). The analysis also highlights significant diagnostic gaps, as 61.2% of cases did not undergo IHC testing due to resource constraints, leading to potential biases in cancer prevalence and diagnostic accuracy. The study emphasizes the complementary role of IHC in confirming ambiguous H&E findings, with strong alignment observed when both methods were used. However, the absence of IHC in many cases limits the robustness of conclusions, suggesting the need for increased access to IHC testing. The findings advocate for integrating IHC into routine diagnostics, expanding diagnostic capabilities, and improving sample sizes to ensure more reliable and comprehensive cancer data.
基金This study was funded by the National Natural Science Foundation of China,No.81470200(to XJR).
文摘Electroacupuncture(EA)has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia.However,there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects.In this study,EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed.Except for the normal group,rats in other groups were fed a high-fat diet throughout the whole experiment.Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks.Middle cerebral artery thrombus models were induced by pasting 50%FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group.EA1 group rats received EA at bilateral ST40(Fenglong)for 7 days before the thrombosis.Rats in the EA1 and EA2 groups received EA at GV20(Baihui)and bilateral ST40 for 14 days after model establishment.Neuronal health was assessed by hematoxylin-eosin staining in the brain.Hyperlipidemia was assessed by biochemical methods that measured total cholesterol,triglyceride,low-density lipoprotein and high-density lipoprotein in blood sera.Behavioral analysis was used to confirm the establishment of the model.Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region.The results demonstrated that,compared with the model group,blood lipid levels significantly decreased,glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups.The repair effect was superior in the EA1 group than in the EA2 group.These findings confirm that EA can reduce blood lipid,inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis.All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine,China(approval No.BUCM-3-2018022802-1002)on April 12,2018.
基金Supported by National Natural Science Foundation of China,No.82074025Scientific Research Project of Heilongjiang Health Committee,No.2020-293Scientific and Technological Innovation Project for College Students of Heilongjiang University of Chinese Medicine,No.2021-13.
文摘BACKGROUND Diabetes is a metabolic disease with a high complication rate.Diabetic foot ulcers(DFUs)seriously affect the quality of life of patients.A total of 15%-20%of diabetic patients develop DFUs,which heal with difficulty over a long time and can result in amputation and disability.Traditional Chinese medicine has a unique effect in the treatment of skin ulcerative diseases.Ruyi Jinhuang powder(RHP)is one of the classic prescriptions in traditional Chinese medicine and is widely used in clinical practice.AIM To verify the ability of RHP to promote wound healing by electron microscopy analysis in animal models and hematoxylin-eosin(HE)staining.The effective components of RHP were extracted and identified by gas chromatography-mass spectrometry(GC-MS),and the obtained chemical components were analyzed by network pharmacology methods to predict its therapeutic mechanism.METHODS Sprague Dawley rats were injected with streptozotocin to establish the DFU model.HE staining was used to observe the wound tissue under an electron microscope.The chemical constituents of RHP were extracted first by supercritical fluid extraction and alcohol extraction,and then,GC-MS and ultra-performance liquid chromatography–MS were used to separately identify the chemical constituents.In addition,the"herb-component-target"link was established through the Traditional Chinese Medicine Systems Pharmacology database to obtain the target information,and the molecular docking of important components and key targets was performed in Discovery Studio software.Cytoscape software was used to visualize and analyze the relationship between the chemical composition,targets and Traditional Chinese Medicine network.RESULTS RHP promoted DFU healing in rats by affecting fibroblasts and nerve cells.A total of 89 chemical components were obtained by GC-MS.Network pharmacological analysis revealed that RHP was associated with 36 targets and 27 pathways in the treatment of DFU,of which the important components were luteolin,trans caryophyllene,ar-turmerone,palmitic acid,methyl palmitate,gallic acid,demethoxycurcumin,berberine,and rheic acid.The key targets were posttranscriptional silencing,topoisomerase II alpha,muscarinic acetylcholine receptor M2,interleukin 6,tumor necrosis factor and retinoic X receptor alpha,and the key pathways were the phosphoinositide 3-kinase-protein kinase B signaling pathway,neuroactive ligand–receptor interactions,and the forkhead box O signaling pathway.CONCLUSION Our results indicated that RHP may play a role in the treatment of DFU through these target pathways by affecting insulin resistance,altering the nervous system and immune system,participating in inflammatory responses and regulating cell proliferation,differentiation and apoptosis through other specific mechanisms.
基金supported by the Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2020D01A13 (to CWW)Chengdu Science and Technology Bureau, No. 2019-YF05-00511-SN (to MT)1.3.5 Project for Disciplines of Excellence, West China Hospital, Sichuan University, Nos. ZY2016102 (to MT), and ZY2016203 (to CY)
文摘According to clinical statistics,the mortality of patients with early brainstem hemorrhage is high.In this study,we established rat models of brainstem hemorrhage by injecting type Ⅶ collagenase into the right basotegmental pontine and investigated the pathological changes of early brainstem hemorrhage using multi-sequence magnetic resonance imaging and histopathological methods.We found that brainstem hematoma gradually formed in the injured rats over the first 3 days and then reduced after 7 days.The edema that occurred was mainly of the vasogenic type.No complete myelin sheath structure was found around the focus of the brainstem hemorrhage.The integrity and continuity of nerve fibers gradually deteriorated over the first 7 days.Neuronal degeneration was mild in the first 3 days and then obviously aggravated on the 7^(th)day.Inflammatory cytokines,interleukin-1β,and tumor necrosis factorαappeared on the 1st day after intracerebral hemorrhage,reached peak levels on the 3^(rd)day,and decreased from the 7^(th)day.Our findings show the characteristics of the progression of early brainstem hemorrhage.
