BACKGROUND The inflammatory response caused by the NLRP3 is closely related to the formation of myocardial ischemiareperfusion injury.Costimulatory receptor CD137 and its ligand play a crucial role in regulating the i...BACKGROUND The inflammatory response caused by the NLRP3 is closely related to the formation of myocardial ischemiareperfusion injury.Costimulatory receptor CD137 and its ligand play a crucial role in regulating the inflammatory immune response in atherosclerosis,which is the fundamental cause of cardiovascular diseases.However,the roles of CD137 signaling in the process of myocardial ischaemia-reperfusion(IR)injury remain unknown.METHODS Genetic ablation was used to determine the functional significance of CD137 in myocardial IR injury.Expression of CD137 was examined by Western-blot,quantitative real-time polymerase chain reaction,and immunohistochemistry in a murine IR model by coronary artery ligation.Even’s blue-TTC staining and echocardiography to evaluate the severity of myocardial IR injury.Furthermore,HL-1 cardiomyocytes treated with agonist-CD137 recombinant protein were used to explore the underlying mechanism in CD137 signaling-induced NLRP3 inflammasome activation in response to hypoxia/reoxygenation or LPS/ATP.RESULTS We demonstrated that CD137 knockout significantly improved cardiac function,accompanied by a markedly reduced NLRP3-mediated inflammatory response and IA/AAR which were reversed by mitophagy inhibitor Mdivi-1.Activating CD137 signaling significantly inhibited mitophagy and provoked NLRP3-mediated inflammatory response in H/R-injured or LPS-primed and ATP-stimulated HL-1 cardiomyocytes,the effects of which could be abolished by either anti-CD137 or mitophagy activator FCCP.Besides,mitochondrial ROS was augmented by activating CD137 signaling through the suppression of mitophagy.CONCLUSIONS Our results reveal that activating CD137 signaling aggravates myocardial IR injury by upregulating NLRP3 inflammasome activation via suppressing mitophagy and promoting mtROS generation.展开更多
Microlens arrays are the key component in the next generation of 3D imaging system, for it exhibits some good optical properties such as extremely large field of view angles, low aberration and distortion, high tempor...Microlens arrays are the key component in the next generation of 3D imaging system, for it exhibits some good optical properties such as extremely large field of view angles, low aberration and distortion, high temporal resolution and infinite depth of field. Although many fabrication methods or processes are proposed for manufacturing such precision component, however, those methods still need to be improved. In this review, those fabrication methods are categorized into direct and indirect method and compared in detail. Two main challenges in manufacturing microlens array are identified: how to obtain a microlens array with good uniformity in a large area and how to produce the microlens array on a curved surface? In order to effectively achieve control of the geometry of a microlens,indirect methods involving the use of 3D molds and replication technologies are suggested. Further development of ultraprecision machining technology is needed to reduce the surface fluctuation by considering the dynamics of machine tool in tool path planning. Finally, the challenges and opportunities of manufacturing microlens array in industry and academic research are discussed and several principle conclusions are drawn.展开更多
AIM: To determine the influence of excision repair cross complementing group 1 (ERCC1 ) codon 118 polymorphism and mRNA level on the clinical outcome of gastric cancer patients treated with oxaliplatin-based adjuvant ...AIM: To determine the influence of excision repair cross complementing group 1 (ERCC1 ) codon 118 polymorphism and mRNA level on the clinical outcome of gastric cancer patients treated with oxaliplatin-based adjuvant chemotherapy. METHODS: Eighty-nine gastric cancer patients treated with oxalipatin-based adjuvant chemotherapy were included in this study. ERCC1 codon 118 C/T polymorphism was tested by polymerase chain reaction-ligation detection reaction (PCR-LDR) method in peripheral blood lymphocytes of those patients; and the intratumoral ERCC1 mRNA expression was measured using reverse transcription PCR in 62 patients whose tumor tissue specimens were available. RESULTS: No significant relationship was found between ERCC1 codon 118 polymorphism and ERCC1 mRNA level. The median relapse-free and overall survival period was 20.1 mo and 28.4 mo, respectively. The relapse-free and overall survivals in patients with low levels of ERCC1 mRNA were significantly longer than those in patients with high levels (P < 0.05), while there was no significant association found between ERCC1 118 genotypes and the disease prognosis. Multivariate analysis also showed that ERCC1 mRNA level was a potential predictor for relapse and survival in gastric cancer patients treated with oxaliplatin-based adjuvant chemotherapy (P < 0.05). CONCLUSION: ERCC1 codon 118 polymorphism has no signifi cant impact on ERCC1 mRNA expression, and the intratumoral ERCC1 mRNA level but not codon 118 polymorphism may be a useful predictive parameter for the relapse and survival of gastric cancer patients receiving oxaliplatin-based adjuvant chemotherapy.展开更多
The histone deacetylase inhibitor, trichostatin A, is used to treat Alzheimer’s disease and can improve learning and memory but its underlying mechanism of action is unknown. To determine whether the therapeutic effe...The histone deacetylase inhibitor, trichostatin A, is used to treat Alzheimer’s disease and can improve learning and memory but its underlying mechanism of action is unknown. To determine whether the therapeutic effect of trichostatin A on Alzheimer’s disease is associated with the nuclear factor erythroid 2-related factor 2(Nrf2) and Kelch-like epichlorohydrin-related protein-1(Keap1) signaling pathway, amyloid β-peptide 25–35(Aβ25–35) was used to induce Alzheimer’s disease-like pathological changes in SH-SY5 Y neuroblastoma cells. Cells were then treated with trichostatin A. The effects of trichostatin A on the expression of Keap1 and Nrf2 were detected by real-time quantitative polymerase chain reaction, western blot assays and immunofluorescence. Total antioxidant capacity and autophagy activity were evaluated by total antioxidant capacity assay kit and light chain 3-I/II levels, respectively. We found that trichostatin A increased cell viability and Nrf2 expression, and decreased Keap1 expression in SH-SY5 Y cells. Furthermore, trichostatin A increased the expression of Nrf2-related target genes, such as superoxide dismutase, NAD(P)H quinone dehydrogenase 1 and glutathione S-transferase, thereby increasing the total antioxidant capacity of SH-SY5 Y cells and inhibiting amyloid β-peptide-induced autophagy. Knockdown of Keap1 in SH-SY5 Y cells further increased trichostatin A-induced Nrf2 expression. These results indicate that the therapeutic effect of trichostatin A on Alzheimer’s disease is associated with the Keap1-Nrf2 pathway. The mechanism for this action may be that trichostatin A increases cell viability and the antioxidant capacity of SH-SY5 Y cells by alleviating Keap1-mediated inhibition Nrf2 signaling, thereby alleviating amyloid β-peptide-induced cell damage.展开更多
AIM: To study the effectiveness and mechanisms of anti-human vascular endothelial growth factor (hVEGF) hairpin ribozyme on angiogenesis,oncogenicity and tumor growth in a hepatocarcinoma cell line and a xenografted m...AIM: To study the effectiveness and mechanisms of anti-human vascular endothelial growth factor (hVEGF) hairpin ribozyme on angiogenesis,oncogenicity and tumor growth in a hepatocarcinoma cell line and a xenografted model. METHODS: The artificial anti-hVEGF hairpin ribozyme was transfected into hepatocarcinoma cell line SMMC-7721 and,subsequently,polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) were performed to confirm the ribozyme gene integration and transcription. To determine the effects of ribozyme ,VEGF expression was detected by semiquantitative RT-PCR and enzyme liked immunosorbent assay (ELISA). MTT assay was carried out to measure the cell proliferation. Furthermore,the transfected and control cells were inoculated into nude mice respectively,the growth of cells in nude mice and angiogenesis were observed. RESULTS: VEGF expression was down-regulated sharply by ribozyme in transfected SMMC-7721 cells and xenografted tumor. Compared to the control group,the transfected cells grew slower in cell cultures and xenografts,and the xenograft formation was delayed as well. In addition,the microvessel density of the xenografted tumor was obviously declined in the transfected group. As demonstratedby microscopy,reduction of VEGF production induced by ribozyme resulted in a significantly higher cell differentiation and less proliferation vigor in xenografted tumor. CONCLUSION: Anti-hVEGF hairpin ribozyme can effectively inhibit VEGF expression and growth of hepatocarcinoma in vitro and in vivo. VEGF is functionally related to cell proliferation,differentiation and tumori-genesis in hepatocarcinoma.展开更多
A new method for the determination of arecoline in Semen Arecae decoction pieces by microchip capillary electrophoresis with contactless conductivity detection(MCE-CCD) was proposed.The effects of various electrophore...A new method for the determination of arecoline in Semen Arecae decoction pieces by microchip capillary electrophoresis with contactless conductivity detection(MCE-CCD) was proposed.The effects of various electrophoretic operating parameters on the analysis of arecoline were studied.Under the optimal conditions,arecoline was rapidly separated and detected in 1 min with good linearity over the concentration range of 20-1500 μM(r2=0.9991) and the detection limit of 5 μM(S/N=3).The method was used for the analysis of arecoline satisfactorily with a recovery of 96.8-104%.展开更多
AIM:To investigate the effects and mechanism ofβ-elemene on the expressions of hypoxia-inducible factor-1α(HIF-lα),vascular endothelial growth factor(VEGF)and inducible nitric oxide synthase(iNOS)in human retinal p...AIM:To investigate the effects and mechanism ofβ-elemene on the expressions of hypoxia-inducible factor-1α(HIF-lα),vascular endothelial growth factor(VEGF)and inducible nitric oxide synthase(iNOS)in human retinal pigment epithelial(RPE)cells under high glucose conditions.METHODS:ARPE-19 cell line was cultured under eight conditions:1)low glucose(LG;5.5 mmol/L);2)high glucose(HG;33 mmol/L);3)high glucose with 20μg/m Lβ-elemene(HG+20 E);4)high glucose with 40μg/m Lβ-elemene(HG+40 E);5)high glucose with SB203590[HG+SB203590,p38-mitogen-activated protein kinase(p38-MAPK)pathway inhibitor];6)high glucose with LY294002[HG+LY294002,phosphoinositide 3-kinase/protein kinase B(PI3 K/Akt)pathway inhibitor];7)high glucose with 40μg/m Lβ-elemene and SB203590(HG+40 E+SB203590);and 8)high glucose with 40μg/m Lβ-elemene and LY294002(HG+40 E+LY294002).Cells were treated in conditions 1-4 for 24 and 48 h,while for 48 h in conditions 5-8.Then m RNA and protein levels of HIF-1α,VEGF and iNOS in cells were measured by real-time polymerase chain reaction(q PCR),immunofluorescence and Western blotting,respectively.Furthermore,protein levels of total p38-MAPK,phosphorylated p38-MAPK(p38-MAPK-P),total Akt and phosphorylated Akt(Akt-P)in cells of conditions 2 and 4 which treated for 48 h were measured by Western blotting.RESULTS:The m RNA levels and protein levels of HIF-1α,VEGF and iNOS in cells were significantly reduced in conditions 3-8 when compared with those in condition 2(P<0.05).These reductions were more obvious in conditions treated for 48 h than in conditions treated for 24 h.The protein levels of p38-MAPK-P and Akt-P in cells of condition 4 were significantly lower than in condition 2(P<0.01).CONCLUSION:β-elemene down-regulates HIF-1α,VEGF and iNOS in ARPE-19 cells under a high glucose condition.The inhibitory effect ofβ-elemene is more significant when its concentration and treatment time are increased,as well as it is combined with SB203590 or LY294002 treatment.P38-MAPK and PI3 K/Akt signaling pathways may play a role in this inhibitory effect.展开更多
Objective:To evaluate the effect of midkine on lipopolysaccharide(LPS)-induced airway smooth muscle cells(ASMCs).Methods:LPS-stimulated acute lung injury model was used to analyze the effect of midkine on ASMCs in vit...Objective:To evaluate the effect of midkine on lipopolysaccharide(LPS)-induced airway smooth muscle cells(ASMCs).Methods:LPS-stimulated acute lung injury model was used to analyze the effect of midkine on ASMCs in vitro.Recombinant midkine and midkine siRNA were used to investigate the role of Notch2 signaling pathway.Cell proliferation was assessed using Cell Counting Kit-8 assay.Additionally,apoptosis was measured by flow cytometry and protein and mRNA expression of midkine and Notch2 was assessed by Western blotting and qPCR,respectively.Immunofluorescence analysis was also conducted.Results:LPS increased the mRNA and protein expression of midkine and Notch2.Midkine silencing reduced LPS-induced midkine and Notch2 expression.In addition,midkine silencing further reduced the viability and increased apoptosis of ASMCs induced by LPS,which was attenuated by recombinant midkine.Conclusions:The midkine/Notch2 signaling pathway plays a regulatory role in ASMC proliferation and apoptosis in airway inflammation.展开更多
AIM: To investigate the roles of PKC-α/ezrin signals in phagocytosis crisis of retinal pigment epithelium(RPE) cells in light damage model. METHODS: Light induced mice RPE injury model was established by continuously...AIM: To investigate the roles of PKC-α/ezrin signals in phagocytosis crisis of retinal pigment epithelium(RPE) cells in light damage model. METHODS: Light induced mice RPE injury model was established by continuously irradiating cool white light at different exposure time(0, 4, 8h light intensity: 4.18×10-6 J/cm2). In vitro, human ARPE-19 cells treated with the doses and intensity(1.57×10-6 J/cm2) of laser irradiation. Histology analysis was evaluated by hematoxylin and eosin(HE) staining. In vivo RPE phagocytosis was quantified by measuring the accumulation of photoreceptor outer segments in the sub-retinal space. In vitro RPE phagocytosis was assessed by calculating the relative fluorescence intensity of FITC-labeled microspheres in ARPE-19 cells. To further investigate the molecular mechanism, the activation of PKC-α/ezrin signal was evaluated by Western blot in vivo and in vitro.RESULTS: HE staining revealed that the thickness of outer nuclear layer decreased significantly after 4 and 8h light exposure. By immunostaining with rhodopsin, a significant greater accumulation of photoreceptor outer segment was noticed after light injury. In vitro, light injuredRPE cells had less phagocytic activity in a dose dependent manner than that of the normal control(P<0.01). Western blot suggested the activation of PKC-α/ezrin signaling was down-regulated in a dose-dependent manner after light exposure. CONCLUSION: Our data suggest that light induced phagocytic crisis of RPE cells may result from the downregulation of PKC-α/ezrin signaling.展开更多
Midkine(MK)is a heparin-bound growth factor.Initially MK was discovered and only expressed during the embryonic period.As the advances of research,MK was found to be expressed in many diseases,such as tumor diseases(p...Midkine(MK)is a heparin-bound growth factor.Initially MK was discovered and only expressed during the embryonic period.As the advances of research,MK was found to be expressed in many diseases,such as tumor diseases(pancreatic cancer,liver cancer,gastric cancer,lung cancer,etc.),acute orchronic inflammatory diseases(ARDS,Chronic renal failure,COPD,etc.),which are related to the molecular structure,signaling pathways and biological effects of MK.Although the signaling pathway mechanism of MK has not been fully elucidated,it is necessary to study its signaling pathway mechanism and play a key role in the treatment of some clinical diseases.This article summarizes the molecular structure of MK and related mechanisms of MK and related diseases,and summarizes how MK participates in the occurrence and development of diseases.And provide a reference for MK may become an important target for disease diagnosis and treatment.展开更多
AIM: To investigate the feasibility of detecting methylated fecal DNA as a screening tool for colorectal carcinoma (CRC) and precancerous lesions. METHODS: Methylated secreted frizzled-related protein gene 2 (SFRP2 ),...AIM: To investigate the feasibility of detecting methylated fecal DNA as a screening tool for colorectal carcinoma (CRC) and precancerous lesions. METHODS: Methylated secreted frizzled-related protein gene 2 (SFRP2 ), hyperplastic polyposis protein gene (HPP1 ) and O6-methylguanine-DNA methyltransferase gene (MGMT) in stools from 52 patients with CRC, 35 patients with benign colorectal diseases and 24 normal individuals were analyzed using methylation-specific PCR. RESULTS: Methylated SFRP2, HPP1 and MGMT were detected in 94.2%, 71.2%, 48.1% of CRC patients and 52.4%, 57.1%, 28.6% of adenoma patients, respectively. The overall prevalence of fecal DNA with at least one methylated gene was 96.2% and 81.8% in patients with CRC and precancerous lesions, respectively. In contrast, only one of the 24 normal individuals revealed methylated DNA. These results indicated a 93.7% sensitivity and a 77.1% specificity of the assay for detecting CRC and precancerous lesions. CONCLUSION: Methylation testing of fecal DNA using a panel of epigenetic markers may be a simple and promising non-invasive screening method for CRC and precancerous lesions.展开更多
The regulatory mechanisms of cytoplasmic Ca2+ after myocardial infarction-induced Ca2+ overload involve secretory pathway Ca2+-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi app...The regulatory mechanisms of cytoplasmic Ca2+ after myocardial infarction-induced Ca2+ overload involve secretory pathway Ca2+-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2+ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2+ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2+ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2+ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2+-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2+-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2+-ATPase 1 can be an important neuroprotective target of ischemic stroke.展开更多
Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open read...Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open reading frame(ORF)of EST sequence from Spirometra mansoni was obtained by ORE finder and was translated into amino acid residue by DNAclub.The structure domain was analyzed by Blast.By the method of online analysis tools:Protparam,InterProScan,protscale.SignalP-3.0,PSORTⅡ,BepiPred,TMHMM,VectorNT1 Suite 9 packages and Phyre2,the structure and function of the protein were predicted and analyzed.Results:The results showed that the EST sequence was Sm TCTP with 173 amino acid residues,theoretical molecular weight was 19 872.0 Da.The protein has the closest evolutionary status with Clonorchis sinensis.Schistosoma mansoni,and Schistosoma japonicum.Then it had no signal peptide site and transmembrane domain.Secondary structure of TCTP contained twoα-helices and eightβ-strands.Conclusions:Sm TCTP was a variety of biological functions of protein that may be used as a vaccine candidate molecule anti drug target.展开更多
AIM: To evaluate the effect of β-elemene on the expressions of hypoxia-inducible factor(HIF)-lα, vascular endothelial growth factor(VEGF) and inducible nitric oxide synthase(i NOS) in a streptozotocin(STZ) induced d...AIM: To evaluate the effect of β-elemene on the expressions of hypoxia-inducible factor(HIF)-lα, vascular endothelial growth factor(VEGF) and inducible nitric oxide synthase(i NOS) in a streptozotocin(STZ) induced diabetic SpragueDawley(SD) rat model.METHODS: SD rats were administered an abdominal injection of STZ and induced to a diabetic model. After 6 wk course of diabetes, the treatment groups were given β-elemene through periocular and intravitreous injection separately and the control groups were given blank emulsion injection. HE staining was used to observe the morphology of retina. The m RNA expressions of HIF-1α, VEGF and i NOS was assayed by real-time polymerase chain reaction(PCR) and the protein expression was measured by Western blot and immunocytochemistry methods.RESULTS: The results indicated that the protein and m RNA expressions of HIF-1α, VEGF and i NOS after treated by β-elemene periocularly and intravitreally injections were all found to be reduced compared with the levels in the diabetic rats group(P<0.05). The inhibitory effect of intravitreal injection was more remarkable.CONCLUSION: The results show β-elemene protect the retina of diabetic rats from high glucose damage by downregulating the expression of HIF-1α, VEGF and iNOS.展开更多
The biaxially textured ion beam-assisted deposited(IBAD)-MgO templates were successfully prepared on polycrystalline Hastelloy metal substrate with reel-to-reel system for YB2Cu3O7-d(YBCO)-coated conductors.By the sol...The biaxially textured ion beam-assisted deposited(IBAD)-MgO templates were successfully prepared on polycrystalline Hastelloy metal substrate with reel-to-reel system for YB2Cu3O7-d(YBCO)-coated conductors.By the solution deposition planarization technique,amorphous Y2O3films were coated on untreated Hastelloy substrate as the bed layer to obtain smooth,dense,and crack-free surface for subsequent IBAD-MgO deposition.The 50 m long IBAD-MgO and homo-epitaxial(epi)-MgO buffer layers deposited on Y2O3films exhibit excellent crystallographic consistency along the scope with full width half maximum(FWHM)values of(110)DU and(200)Dx in the range of 5.5°–6.0°and3.0°–3.5°,respectively.To match the lattice constant of YBCO material,LaMnO3cap layer was fabricated on IBAD-MgO templates by radio frequency(rf)magnetron sputtering with the FWHM values of in-plane and out-ofplane of 6.8°and 3.7°,respectively,indicating excellent biaxial texture.展开更多
The epidemic caused by the new coronavirus infection is a global public health emergency. The new coronavirus pneumonia (COVID-19) has caused more deaths in China than the SARS-CoV outbreak in 2003. Since the virus ha...The epidemic caused by the new coronavirus infection is a global public health emergency. The new coronavirus pneumonia (COVID-19) has caused more deaths in China than the SARS-CoV outbreak in 2003. Since the virus has a high homology with SARS-CoV, it is speculates that it may infect cells through the S protein-ACE2 pathway then cause tracheal epithelial cell damage and alveolar damage, further induce inflammatory factor storm and trigger apoptosis cascade reaction and other mechanisms leading to acute lung injury. At early stage, the new coronavirus pneumonia mainly presents with non-specific manifestations of fatigue, fever, cough, etc., Then the respiratory tract will be involved, and evolved into the new type of infectious disease with lung lesions. Under radiology images, ground glass-like changes can be observed in the lungs. With its aggravation, progressive hypoxemia, acute respiratory distress syndrome, and even death occurs. At present, the correlation between the new coronavirus pneumonia and acute lung injury is not yet clear. This article reviews the characteristics of the new coronavirus, the possible mechanism of acute lung injury in new coronavirus pneumonia, post-infection diagnosis and current treatment options in order to provide evidence for its treatment.展开更多
Paraquat is a bipyridine dichloride non-selective herbicide,which was widely used in the world in the last century.Now,paraquat is banned in most countries because of the extremely high lethality and the lack of speci...Paraquat is a bipyridine dichloride non-selective herbicide,which was widely used in the world in the last century.Now,paraquat is banned in most countries because of the extremely high lethality and the lack of specific detoxification drugs.However,death due to paraquat poisoning still occurs frequently,thus it is of great clinical significance to explore the molecular mechanism of paraquat poisoning and the detoxification drugs.Paraquat poisoning causes multiple dysfunction of the lung,liver,kidney,heart,and brain through complex molecular mechanisms.About the mechanism there are excessive inflammatory reaction theory,REDOX reaction imbalance theory,oxidative stress free radical damage theory,calcium overload theory,NO molecular damage and cell apoptosis theory,etc.For the treatment of paraquat poisoning,paraquat antibody,pathway target blocker and related factor antibody have been developed in recent years.Although certain effects have been achieved,the treatment efficiency has not been significantly improved.This paper summarized the mechanism of signal transduction pathways involved in lung injury induced by paraquat poisoning in order to provide a theoretical basis for further research.展开更多
基金supported as follows:National Natural Science Foundation of China(81970379)Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX22_3712)Medical Innovation Team Project of Jiangsu Province(CXTDA2017010).
文摘BACKGROUND The inflammatory response caused by the NLRP3 is closely related to the formation of myocardial ischemiareperfusion injury.Costimulatory receptor CD137 and its ligand play a crucial role in regulating the inflammatory immune response in atherosclerosis,which is the fundamental cause of cardiovascular diseases.However,the roles of CD137 signaling in the process of myocardial ischaemia-reperfusion(IR)injury remain unknown.METHODS Genetic ablation was used to determine the functional significance of CD137 in myocardial IR injury.Expression of CD137 was examined by Western-blot,quantitative real-time polymerase chain reaction,and immunohistochemistry in a murine IR model by coronary artery ligation.Even’s blue-TTC staining and echocardiography to evaluate the severity of myocardial IR injury.Furthermore,HL-1 cardiomyocytes treated with agonist-CD137 recombinant protein were used to explore the underlying mechanism in CD137 signaling-induced NLRP3 inflammasome activation in response to hypoxia/reoxygenation or LPS/ATP.RESULTS We demonstrated that CD137 knockout significantly improved cardiac function,accompanied by a markedly reduced NLRP3-mediated inflammatory response and IA/AAR which were reversed by mitophagy inhibitor Mdivi-1.Activating CD137 signaling significantly inhibited mitophagy and provoked NLRP3-mediated inflammatory response in H/R-injured or LPS-primed and ATP-stimulated HL-1 cardiomyocytes,the effects of which could be abolished by either anti-CD137 or mitophagy activator FCCP.Besides,mitochondrial ROS was augmented by activating CD137 signaling through the suppression of mitophagy.CONCLUSIONS Our results reveal that activating CD137 signaling aggravates myocardial IR injury by upregulating NLRP3 inflammasome activation via suppressing mitophagy and promoting mtROS generation.
基金Supported by Shenzhen Science,Technology and Innovation Commission of China(Grant No.JCYJ20150630115257902)the Research Grants Council of the Hong Kong Special Administrative Region of China(Grant No.ITS/339/13FX)Research Committee of The Hong Kong Polytechnic University,China (Grant No.RUK0)
文摘Microlens arrays are the key component in the next generation of 3D imaging system, for it exhibits some good optical properties such as extremely large field of view angles, low aberration and distortion, high temporal resolution and infinite depth of field. Although many fabrication methods or processes are proposed for manufacturing such precision component, however, those methods still need to be improved. In this review, those fabrication methods are categorized into direct and indirect method and compared in detail. Two main challenges in manufacturing microlens array are identified: how to obtain a microlens array with good uniformity in a large area and how to produce the microlens array on a curved surface? In order to effectively achieve control of the geometry of a microlens,indirect methods involving the use of 3D molds and replication technologies are suggested. Further development of ultraprecision machining technology is needed to reduce the surface fluctuation by considering the dynamics of machine tool in tool path planning. Finally, the challenges and opportunities of manufacturing microlens array in industry and academic research are discussed and several principle conclusions are drawn.
基金Supported by A Grant From Scientif ic and Technologic Bureau of Wuxi, CLZ00612
文摘AIM: To determine the influence of excision repair cross complementing group 1 (ERCC1 ) codon 118 polymorphism and mRNA level on the clinical outcome of gastric cancer patients treated with oxaliplatin-based adjuvant chemotherapy. METHODS: Eighty-nine gastric cancer patients treated with oxalipatin-based adjuvant chemotherapy were included in this study. ERCC1 codon 118 C/T polymorphism was tested by polymerase chain reaction-ligation detection reaction (PCR-LDR) method in peripheral blood lymphocytes of those patients; and the intratumoral ERCC1 mRNA expression was measured using reverse transcription PCR in 62 patients whose tumor tissue specimens were available. RESULTS: No significant relationship was found between ERCC1 codon 118 polymorphism and ERCC1 mRNA level. The median relapse-free and overall survival period was 20.1 mo and 28.4 mo, respectively. The relapse-free and overall survivals in patients with low levels of ERCC1 mRNA were significantly longer than those in patients with high levels (P < 0.05), while there was no significant association found between ERCC1 118 genotypes and the disease prognosis. Multivariate analysis also showed that ERCC1 mRNA level was a potential predictor for relapse and survival in gastric cancer patients treated with oxaliplatin-based adjuvant chemotherapy (P < 0.05). CONCLUSION: ERCC1 codon 118 polymorphism has no signifi cant impact on ERCC1 mRNA expression, and the intratumoral ERCC1 mRNA level but not codon 118 polymorphism may be a useful predictive parameter for the relapse and survival of gastric cancer patients receiving oxaliplatin-based adjuvant chemotherapy.
基金Supported by The South China National Research Centre for Integrated Biosciences, and has been conducted collaboratively between Zhongshan University (ZU)The Chinese University of Hong Kong
文摘The histone deacetylase inhibitor, trichostatin A, is used to treat Alzheimer’s disease and can improve learning and memory but its underlying mechanism of action is unknown. To determine whether the therapeutic effect of trichostatin A on Alzheimer’s disease is associated with the nuclear factor erythroid 2-related factor 2(Nrf2) and Kelch-like epichlorohydrin-related protein-1(Keap1) signaling pathway, amyloid β-peptide 25–35(Aβ25–35) was used to induce Alzheimer’s disease-like pathological changes in SH-SY5 Y neuroblastoma cells. Cells were then treated with trichostatin A. The effects of trichostatin A on the expression of Keap1 and Nrf2 were detected by real-time quantitative polymerase chain reaction, western blot assays and immunofluorescence. Total antioxidant capacity and autophagy activity were evaluated by total antioxidant capacity assay kit and light chain 3-I/II levels, respectively. We found that trichostatin A increased cell viability and Nrf2 expression, and decreased Keap1 expression in SH-SY5 Y cells. Furthermore, trichostatin A increased the expression of Nrf2-related target genes, such as superoxide dismutase, NAD(P)H quinone dehydrogenase 1 and glutathione S-transferase, thereby increasing the total antioxidant capacity of SH-SY5 Y cells and inhibiting amyloid β-peptide-induced autophagy. Knockdown of Keap1 in SH-SY5 Y cells further increased trichostatin A-induced Nrf2 expression. These results indicate that the therapeutic effect of trichostatin A on Alzheimer’s disease is associated with the Keap1-Nrf2 pathway. The mechanism for this action may be that trichostatin A increases cell viability and the antioxidant capacity of SH-SY5 Y cells by alleviating Keap1-mediated inhibition Nrf2 signaling, thereby alleviating amyloid β-peptide-induced cell damage.
基金Supported by the Grant from Calling for Tenders by Key Subject of Jiangsu Province, No. WK200221
文摘AIM: To study the effectiveness and mechanisms of anti-human vascular endothelial growth factor (hVEGF) hairpin ribozyme on angiogenesis,oncogenicity and tumor growth in a hepatocarcinoma cell line and a xenografted model. METHODS: The artificial anti-hVEGF hairpin ribozyme was transfected into hepatocarcinoma cell line SMMC-7721 and,subsequently,polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) were performed to confirm the ribozyme gene integration and transcription. To determine the effects of ribozyme ,VEGF expression was detected by semiquantitative RT-PCR and enzyme liked immunosorbent assay (ELISA). MTT assay was carried out to measure the cell proliferation. Furthermore,the transfected and control cells were inoculated into nude mice respectively,the growth of cells in nude mice and angiogenesis were observed. RESULTS: VEGF expression was down-regulated sharply by ribozyme in transfected SMMC-7721 cells and xenografted tumor. Compared to the control group,the transfected cells grew slower in cell cultures and xenografts,and the xenograft formation was delayed as well. In addition,the microvessel density of the xenografted tumor was obviously declined in the transfected group. As demonstratedby microscopy,reduction of VEGF production induced by ribozyme resulted in a significantly higher cell differentiation and less proliferation vigor in xenografted tumor. CONCLUSION: Anti-hVEGF hairpin ribozyme can effectively inhibit VEGF expression and growth of hepatocarcinoma in vitro and in vivo. VEGF is functionally related to cell proliferation,differentiation and tumori-genesis in hepatocarcinoma.
基金support from the National Natural Science Foundation of China (Nos. 20727006,21075139)Guangdong Provincial Science and Technology Project (2008A030102009)the Medical Scientific Research Foundation of Guangdong Province(A2012155)
文摘A new method for the determination of arecoline in Semen Arecae decoction pieces by microchip capillary electrophoresis with contactless conductivity detection(MCE-CCD) was proposed.The effects of various electrophoretic operating parameters on the analysis of arecoline were studied.Under the optimal conditions,arecoline was rapidly separated and detected in 1 min with good linearity over the concentration range of 20-1500 μM(r2=0.9991) and the detection limit of 5 μM(S/N=3).The method was used for the analysis of arecoline satisfactorily with a recovery of 96.8-104%.
文摘AIM:To investigate the effects and mechanism ofβ-elemene on the expressions of hypoxia-inducible factor-1α(HIF-lα),vascular endothelial growth factor(VEGF)and inducible nitric oxide synthase(iNOS)in human retinal pigment epithelial(RPE)cells under high glucose conditions.METHODS:ARPE-19 cell line was cultured under eight conditions:1)low glucose(LG;5.5 mmol/L);2)high glucose(HG;33 mmol/L);3)high glucose with 20μg/m Lβ-elemene(HG+20 E);4)high glucose with 40μg/m Lβ-elemene(HG+40 E);5)high glucose with SB203590[HG+SB203590,p38-mitogen-activated protein kinase(p38-MAPK)pathway inhibitor];6)high glucose with LY294002[HG+LY294002,phosphoinositide 3-kinase/protein kinase B(PI3 K/Akt)pathway inhibitor];7)high glucose with 40μg/m Lβ-elemene and SB203590(HG+40 E+SB203590);and 8)high glucose with 40μg/m Lβ-elemene and LY294002(HG+40 E+LY294002).Cells were treated in conditions 1-4 for 24 and 48 h,while for 48 h in conditions 5-8.Then m RNA and protein levels of HIF-1α,VEGF and iNOS in cells were measured by real-time polymerase chain reaction(q PCR),immunofluorescence and Western blotting,respectively.Furthermore,protein levels of total p38-MAPK,phosphorylated p38-MAPK(p38-MAPK-P),total Akt and phosphorylated Akt(Akt-P)in cells of conditions 2 and 4 which treated for 48 h were measured by Western blotting.RESULTS:The m RNA levels and protein levels of HIF-1α,VEGF and iNOS in cells were significantly reduced in conditions 3-8 when compared with those in condition 2(P<0.05).These reductions were more obvious in conditions treated for 48 h than in conditions treated for 24 h.The protein levels of p38-MAPK-P and Akt-P in cells of condition 4 were significantly lower than in condition 2(P<0.01).CONCLUSION:β-elemene down-regulates HIF-1α,VEGF and iNOS in ARPE-19 cells under a high glucose condition.The inhibitory effect ofβ-elemene is more significant when its concentration and treatment time are increased,as well as it is combined with SB203590 or LY294002 treatment.P38-MAPK and PI3 K/Akt signaling pathways may play a role in this inhibitory effect.
基金supported by the National Natural Science Foundation of China(NO.81660011,81960351)Hainan Provincial Social Development Foundation(NO.ZDYFXGFY2020004)Hainan Provincial Medical and Health Research Project(NO.22A200036).
文摘Objective:To evaluate the effect of midkine on lipopolysaccharide(LPS)-induced airway smooth muscle cells(ASMCs).Methods:LPS-stimulated acute lung injury model was used to analyze the effect of midkine on ASMCs in vitro.Recombinant midkine and midkine siRNA were used to investigate the role of Notch2 signaling pathway.Cell proliferation was assessed using Cell Counting Kit-8 assay.Additionally,apoptosis was measured by flow cytometry and protein and mRNA expression of midkine and Notch2 was assessed by Western blotting and qPCR,respectively.Immunofluorescence analysis was also conducted.Results:LPS increased the mRNA and protein expression of midkine and Notch2.Midkine silencing reduced LPS-induced midkine and Notch2 expression.In addition,midkine silencing further reduced the viability and increased apoptosis of ASMCs induced by LPS,which was attenuated by recombinant midkine.Conclusions:The midkine/Notch2 signaling pathway plays a regulatory role in ASMC proliferation and apoptosis in airway inflammation.
基金Supported by the National Natural Science Foundation of China(No.81641057)the Natural Science Foundation of Liaoning Province(No.201602292No.201602298)
文摘AIM: To investigate the roles of PKC-α/ezrin signals in phagocytosis crisis of retinal pigment epithelium(RPE) cells in light damage model. METHODS: Light induced mice RPE injury model was established by continuously irradiating cool white light at different exposure time(0, 4, 8h light intensity: 4.18×10-6 J/cm2). In vitro, human ARPE-19 cells treated with the doses and intensity(1.57×10-6 J/cm2) of laser irradiation. Histology analysis was evaluated by hematoxylin and eosin(HE) staining. In vivo RPE phagocytosis was quantified by measuring the accumulation of photoreceptor outer segments in the sub-retinal space. In vitro RPE phagocytosis was assessed by calculating the relative fluorescence intensity of FITC-labeled microspheres in ARPE-19 cells. To further investigate the molecular mechanism, the activation of PKC-α/ezrin signal was evaluated by Western blot in vivo and in vitro.RESULTS: HE staining revealed that the thickness of outer nuclear layer decreased significantly after 4 and 8h light exposure. By immunostaining with rhodopsin, a significant greater accumulation of photoreceptor outer segment was noticed after light injury. In vitro, light injuredRPE cells had less phagocytic activity in a dose dependent manner than that of the normal control(P<0.01). Western blot suggested the activation of PKC-α/ezrin signaling was down-regulated in a dose-dependent manner after light exposure. CONCLUSION: Our data suggest that light induced phagocytic crisis of RPE cells may result from the downregulation of PKC-α/ezrin signaling.
基金National Natural Science Foundation of China(No.81660011)
文摘Midkine(MK)is a heparin-bound growth factor.Initially MK was discovered and only expressed during the embryonic period.As the advances of research,MK was found to be expressed in many diseases,such as tumor diseases(pancreatic cancer,liver cancer,gastric cancer,lung cancer,etc.),acute orchronic inflammatory diseases(ARDS,Chronic renal failure,COPD,etc.),which are related to the molecular structure,signaling pathways and biological effects of MK.Although the signaling pathway mechanism of MK has not been fully elucidated,it is necessary to study its signaling pathway mechanism and play a key role in the treatment of some clinical diseases.This article summarizes the molecular structure of MK and related mechanisms of MK and related diseases,and summarizes how MK participates in the occurrence and development of diseases.And provide a reference for MK may become an important target for disease diagnosis and treatment.
基金grant from Scientific and Technologic Bureau of Wuxi, No. CS055010
文摘AIM: To investigate the feasibility of detecting methylated fecal DNA as a screening tool for colorectal carcinoma (CRC) and precancerous lesions. METHODS: Methylated secreted frizzled-related protein gene 2 (SFRP2 ), hyperplastic polyposis protein gene (HPP1 ) and O6-methylguanine-DNA methyltransferase gene (MGMT) in stools from 52 patients with CRC, 35 patients with benign colorectal diseases and 24 normal individuals were analyzed using methylation-specific PCR. RESULTS: Methylated SFRP2, HPP1 and MGMT were detected in 94.2%, 71.2%, 48.1% of CRC patients and 52.4%, 57.1%, 28.6% of adenoma patients, respectively. The overall prevalence of fecal DNA with at least one methylated gene was 96.2% and 81.8% in patients with CRC and precancerous lesions, respectively. In contrast, only one of the 24 normal individuals revealed methylated DNA. These results indicated a 93.7% sensitivity and a 77.1% specificity of the assay for detecting CRC and precancerous lesions. CONCLUSION: Methylation testing of fecal DNA using a panel of epigenetic markers may be a simple and promising non-invasive screening method for CRC and precancerous lesions.
基金supported by the National Natural Science Foundation of China,No.81171239
文摘The regulatory mechanisms of cytoplasmic Ca2+ after myocardial infarction-induced Ca2+ overload involve secretory pathway Ca2+-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2+ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2+ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2+ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2+ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2+-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2+-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2+-ATPase 1 can be an important neuroprotective target of ischemic stroke.
基金supported by Natural Science Foundation of China(Foundation No.81260254)
文摘Objective:To predict structure and function of translationally controlled tumor protein(TCTP)from Spirometraa mansoni by bioinformatics technology,and to provide a theoretical basis for further study.Methods:Open reading frame(ORF)of EST sequence from Spirometra mansoni was obtained by ORE finder and was translated into amino acid residue by DNAclub.The structure domain was analyzed by Blast.By the method of online analysis tools:Protparam,InterProScan,protscale.SignalP-3.0,PSORTⅡ,BepiPred,TMHMM,VectorNT1 Suite 9 packages and Phyre2,the structure and function of the protein were predicted and analyzed.Results:The results showed that the EST sequence was Sm TCTP with 173 amino acid residues,theoretical molecular weight was 19 872.0 Da.The protein has the closest evolutionary status with Clonorchis sinensis.Schistosoma mansoni,and Schistosoma japonicum.Then it had no signal peptide site and transmembrane domain.Secondary structure of TCTP contained twoα-helices and eightβ-strands.Conclusions:Sm TCTP was a variety of biological functions of protein that may be used as a vaccine candidate molecule anti drug target.
文摘AIM: To evaluate the effect of β-elemene on the expressions of hypoxia-inducible factor(HIF)-lα, vascular endothelial growth factor(VEGF) and inducible nitric oxide synthase(i NOS) in a streptozotocin(STZ) induced diabetic SpragueDawley(SD) rat model.METHODS: SD rats were administered an abdominal injection of STZ and induced to a diabetic model. After 6 wk course of diabetes, the treatment groups were given β-elemene through periocular and intravitreous injection separately and the control groups were given blank emulsion injection. HE staining was used to observe the morphology of retina. The m RNA expressions of HIF-1α, VEGF and i NOS was assayed by real-time polymerase chain reaction(PCR) and the protein expression was measured by Western blot and immunocytochemistry methods.RESULTS: The results indicated that the protein and m RNA expressions of HIF-1α, VEGF and i NOS after treated by β-elemene periocularly and intravitreally injections were all found to be reduced compared with the levels in the diabetic rats group(P<0.05). The inhibitory effect of intravitreal injection was more remarkable.CONCLUSION: The results show β-elemene protect the retina of diabetic rats from high glucose damage by downregulating the expression of HIF-1α, VEGF and iNOS.
基金supported by the Sichuan Youth Science and Technology Innovation Research Team (No. 2011JTD0006)Fundamental Research Funds for the Central Universities (Nos. ZYGX2012J039 and ZYGX2011Z002)
文摘The biaxially textured ion beam-assisted deposited(IBAD)-MgO templates were successfully prepared on polycrystalline Hastelloy metal substrate with reel-to-reel system for YB2Cu3O7-d(YBCO)-coated conductors.By the solution deposition planarization technique,amorphous Y2O3films were coated on untreated Hastelloy substrate as the bed layer to obtain smooth,dense,and crack-free surface for subsequent IBAD-MgO deposition.The 50 m long IBAD-MgO and homo-epitaxial(epi)-MgO buffer layers deposited on Y2O3films exhibit excellent crystallographic consistency along the scope with full width half maximum(FWHM)values of(110)DU and(200)Dx in the range of 5.5°–6.0°and3.0°–3.5°,respectively.To match the lattice constant of YBCO material,LaMnO3cap layer was fabricated on IBAD-MgO templates by radio frequency(rf)magnetron sputtering with the FWHM values of in-plane and out-ofplane of 6.8°and 3.7°,respectively,indicating excellent biaxial texture.
基金AHQDS study on the treatment of paraquat poisoning(ZDYF2019125)Midkine on the proliferation of airway smooth muscle cels in SD rat COPD model and its molecular mechanism(81660011)CAMS Innovation Fund for Medical Sciences(2019-I2M-5-023)
文摘The epidemic caused by the new coronavirus infection is a global public health emergency. The new coronavirus pneumonia (COVID-19) has caused more deaths in China than the SARS-CoV outbreak in 2003. Since the virus has a high homology with SARS-CoV, it is speculates that it may infect cells through the S protein-ACE2 pathway then cause tracheal epithelial cell damage and alveolar damage, further induce inflammatory factor storm and trigger apoptosis cascade reaction and other mechanisms leading to acute lung injury. At early stage, the new coronavirus pneumonia mainly presents with non-specific manifestations of fatigue, fever, cough, etc., Then the respiratory tract will be involved, and evolved into the new type of infectious disease with lung lesions. Under radiology images, ground glass-like changes can be observed in the lungs. With its aggravation, progressive hypoxemia, acute respiratory distress syndrome, and even death occurs. At present, the correlation between the new coronavirus pneumonia and acute lung injury is not yet clear. This article reviews the characteristics of the new coronavirus, the possible mechanism of acute lung injury in new coronavirus pneumonia, post-infection diagnosis and current treatment options in order to provide evidence for its treatment.
基金Natural Science Foundation of China-Regional Project(No.81960351)Social development key project of Hainan province(No.ZDYF2019125).
文摘Paraquat is a bipyridine dichloride non-selective herbicide,which was widely used in the world in the last century.Now,paraquat is banned in most countries because of the extremely high lethality and the lack of specific detoxification drugs.However,death due to paraquat poisoning still occurs frequently,thus it is of great clinical significance to explore the molecular mechanism of paraquat poisoning and the detoxification drugs.Paraquat poisoning causes multiple dysfunction of the lung,liver,kidney,heart,and brain through complex molecular mechanisms.About the mechanism there are excessive inflammatory reaction theory,REDOX reaction imbalance theory,oxidative stress free radical damage theory,calcium overload theory,NO molecular damage and cell apoptosis theory,etc.For the treatment of paraquat poisoning,paraquat antibody,pathway target blocker and related factor antibody have been developed in recent years.Although certain effects have been achieved,the treatment efficiency has not been significantly improved.This paper summarized the mechanism of signal transduction pathways involved in lung injury induced by paraquat poisoning in order to provide a theoretical basis for further research.
