BACKGROUND Photodynamic therapy(PDT)is a minimally invasive form of cancer therapy,and the development of a novel photosensitizer(PS)with optimal properties is important for enhancing PDT efficacy.Folate receptor(FR)m...BACKGROUND Photodynamic therapy(PDT)is a minimally invasive form of cancer therapy,and the development of a novel photosensitizer(PS)with optimal properties is important for enhancing PDT efficacy.Folate receptor(FR)membrane protein is frequently overexpressed in 40%of human cancer and a good candidate for tumor-specific targeting.Specific active targeting of PS to FR can be achieved by conjugation with the folate moiety.A folate-linked,near-infrared(NIR)-sensitive probe,folate-Si-rhodamine-1(FolateSiR-1),was previously developed and is expected to be applicable to NIR-PDT.AIM To investigate the therapeutic efficacy of NIR-PDT induced by FolateSiR-1,a FRtargeted PS,in preclinical cancer models.METHODS FolateSiR-1 was developed by conjugating a folate moiety to the Si-rhodamine derivative through a negatively charged tripeptide linker.FR expression in the designated cell lines was examined by western blotting(WB).The selective binding of FolateSiR-1 to FR was confirmed in FR overexpressing KB cells(FR+)and tumors by fluorescence microscopy and in vivo fluorescence imaging.Low FR expressing OVCAR-3 and A4 cell lines were used as negative controls(FR-).The NIR light(635±3 nm)-induced phototoxic effect of FolateSiR-1 was evaluated by cell viability imaging assays.The time-dependent distribution of FolateSiR-1 and its specific accumulation in KB tumors was determined using in vivo longitudinal fluorescence imaging.The PDT effect of FolateSiR-1 was evaluated in KB tumor bearing mice divided into four experimental groups:(1)FolateSiR-1(100μmol/L)alone;(2)FolateSiR-1(100μmol/L)followed by NIR irradiation(50 J/cm2);(3)NIR irradiation(50 J/cm2)alone;and(4)no treatment.Tumor volume measurement and immunohistochemical(IHC)and histological examinations of the tumors were performed to analyze the effect of PDT.RESULTS High FR expression was observed in the KB cells by WB,but not in the OVCAR-3 and A4 cells.Substantial FR-specific binding of FolateSiR-1 was observed by in vitro and in vivo fluorescence imaging.Cell viability imaging assays showed that NIR-PDT induced cell death in KB cells.In vivo longitudinal fluorescence imaging showed rapid peak accumulation of FolateSiR-1 in the KB tumors 2 h after injection.In vivo PDT conducted at this time point caused tumor growth delay.The relative tumor volumes in the PDT group were significantly reduced compared to those in the other groups[5.81±1.74(NIR-PDT)vs 12.24±2.48(Folate-SiR-1),vs 11.84±3.67(IR),vs 12.98±2.78(Untreated),at Day 16,P<0.05].IHC analysis revealed reduced proliferation marker Ki-67-positive cells in the PDT treated tumors,and hematoxylin-eosin staining revealed features of necrotic-and apoptotic cell death.CONCLUSION FolateSiR-1 has potential for use in PDT,and FR-targeted NIR-PDT may open a new effective strategy for the treatment of FR-overexpressing tumors.展开更多
Objective: To investigate the effects of linezolid and moxifloxacin combined with quadruple antituberculosis drugs on CSF cytology, NSE (neuronal specific enolase), NGF(nerve growth facor) and its receptors, endotheli...Objective: To investigate the effects of linezolid and moxifloxacin combined with quadruple antituberculosis drugs on CSF cytology, NSE (neuronal specific enolase), NGF(nerve growth facor) and its receptors, endothelin and its receptors and CRP (C reaction protein) in patients with refractory tuberculous meningitis. Method: A total of 56 patients were selected with tuberculous meningitis in our hospital from February 2014 to December 2017,randomly divided them into 2 groups, each group was 28 cases, set as the observation group and the control group, both groups were treated with quadruple antituberculosis drugs, the observation group was given ilinezolid on this basis, and the control group was combined with moxifloxacin, The course of treatment was 4 weeks, compared the levels of CSF cytology, NSE, NGF and NGF receptors, endothelin and endothelin receptors, and CRP after treatment in the two groups. Result:The CSF cytology, NSE, NGF and NGF receptors, endothelin and endothelin receptor, and CRP levels remained unchanged before treatment, the difference was not statistically significant. After treatment, the chloride and glucose levels in the observation group were higher than those before treatment and that of the control group, the protein, white blood cell count, and cerebrospinal fluid pressure levels were lower than before treatment and that of the control group, the difference was statistically significant;The NSE level in the observation group after treatment was lower than before treatment and that of the control group, the difference was statistically significant;After treatment, the levels of NGF and its receptors in the observation group were higher than those before treatment and that of the control group, and the levels of endothelin, and its receptor, CRP were lower than those before treatment and that of the control group, the difference was statistically significant. Conclusion:The use of linezolid in combination with quadruple antituberculosis drugs to treat refractory tuberculous meningitis has better clinical effect, effectively improve cerebrospinal fluid cytology, regulate cerebrospinal fluid NSE levels, restore NGF, endothelin and its receptor function, reduce inflammatory response, recommended for clinical promotion and application.展开更多
Autism spectrum disorder(ASD)is a serious neurodevelopmental disorder,the etiology and mechanism of which are not yet clear.Studies have shown that folate deficiency can lead to abnormalities in the de-velopment of th...Autism spectrum disorder(ASD)is a serious neurodevelopmental disorder,the etiology and mechanism of which are not yet clear.Studies have shown that folate deficiency can lead to abnormalities in the de-velopment of the central nervous system.Patients with autism spectrum disorders develop folate-alpha recep-tor autoantibodies.Folate-alpha receptor autoantibodies block folate transport,leading to a deficiency of folate in nerve tissues.Folate is effective in treating patients with folate-alpha receptor autoantibodies.展开更多
Invasive nonfunctional pituitary adenomas （NFPAs） are difficult to completely resect and often develop tumor recurrence after initial surgery. Currently, no medications are clinically effective in the control of NFP...Invasive nonfunctional pituitary adenomas （NFPAs） are difficult to completely resect and often develop tumor recurrence after initial surgery. Currently, no medications are clinically effective in the control of NFPA. Although radiation therapy and radiosurgery are useful to prevent tumor regrowth, they are frequently withheld because of severe complications. Boron neutron capture therapy （BNCT） is a binary radiotherapy that selectively and maximally damages tumor cells without harming the surrounding normal tissue. Folate receptor （FR）-targeted boron-10 containing carbon nanoparticles is a novel boron delivery agent that can be selectively taken up by FR-expressing cells via FR-mediated endocytosis. In this study, FR-targeted boron-10 containing carbon nanoparticles were selectively taken up by NFPAs cells expressing FR but not other types of non-FR expressing pituitary adenomas. After incubation with boron-10 containing carbon nanoparticles and following irradiation with thermal neutrons, the cell viability of NFPAs was significantly decreased, while apoptotic cells were simultaneously increased. However, cells administered the same dose of FR-targeted boron-10 containing carbon nanoparticles without neutron irradiation or received the same neutron irradiation alone did not show significant decrease in cell viability or increase in apoptotic cells. The expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated in NFPAs after treatment with FR-mediated BNCT. In conclusion, FR-targeted boron-10 containing carbon nanoparticles may be an ideal delivery system of boron to NFPAs ceils for BNCT. Furthermore, our study also provides a novel insight into therapeutic strategies for invasive NFPA refractory to conventional therapy, while exploring these new applications of BNCT for tumors, especially benign tumors.展开更多
Vulnerable atherosclerotic plaques are responsible for most cardiovascular diseases(CVDs).Folate receptor(FR)positive activated macrophages were thought to be a prominent component in the development of vulnerable pla...Vulnerable atherosclerotic plaques are responsible for most cardiovascular diseases(CVDs).Folate receptor(FR)positive activated macrophages were thought to be a prominent component in the development of vulnerable plaque.The objective of this study is to develop folate conjugated two-dimensional(2D)Pd@Au nanomaterials(Pd@Au-PEG-FA)for targeted multimodal imaging of the FRs in advanced atherosclerotic plaques.Pharmacokinetic and imaging studies(single photon emission computed tomography(SPECT),computed tomography(CT)and photoacoustic(PA)imaging)were performed to confirm the prolonged blood half-life and enrichment of radioactivity in atherosclerotic plaques.Strong signals were detected in vivo with SPECT,CT and PA imaging in heavy atherosclerotic plaques,which were significantly higher than those of the normal aortas after injection of Pd@Au-PEG-FA.Blocking studies with preinjection of excess FA could effectively reduce the targeting ability of Pd@Au-PEG-FA in atherosclerotic plaques,further demonstrating the specific binding of Pd@Au-PEG-FA for plaque lesions.Histopathological characterization revealed that the signal of probe was in accordance with the high-risk plaques.In summary,the Pd@Au-PEG-FA has favorable pharmacokinetic properties and provides a valuable approach for detecting high-risk plaques in the presence of FRs in atherosclerotic plaques.展开更多
The detection of protein/small molecule interactions plays important roles in drug discovery and protein/metabolite interactions in biology. In this work, by coupling the terminal protection of small molecule-linked s...The detection of protein/small molecule interactions plays important roles in drug discovery and protein/metabolite interactions in biology. In this work, by coupling the terminal protection of small molecule-linked ss DNA strategy with the unmodified and positively charged gold nanoparticle((+)Au NP) nanoprobes, we have developed a sensitive and simple colorimetric sensor for the detection of folate receptor, a highly expressed protein in many kinds of malignant tumors. The target folate receptor binds the folate moieties of the folate-linked ss DNA through high affinity interactions and protects the protein-bound ss DNA from digestion by exonuclease I. The protected ss DNA thus adsorbs the((+)Au NP) through electrostatic interactions, leading to a red-to-blue color change of the sensing solution for sensitive colorimetric detection of folate receptor at the sub-nanomolar level. Besides, this colorimetric sensor shows high selectivity toward folate receptor against other control proteins. The developed sensor avoids the modification/conjugation of the Au NP nanoprobes and the involvement of any expensive instruments for signal transduction in protein detection. Featured with these obvious advantages, the colorimetric sensor strategy demonstrated herein can be easily expanded for sensitive and convenient detection of various protein/small molecule interactions.展开更多
To assess targeting of an epothilone folate conjugate(BMS-753493) to the folate receptor(FR)-overexpressed tumor in mice bearing both FRt and FR– tumors,a series of experiments were conducted by quantitative whole-bo...To assess targeting of an epothilone folate conjugate(BMS-753493) to the folate receptor(FR)-overexpressed tumor in mice bearing both FRt and FR– tumors,a series of experiments were conducted by quantitative whole-body autoradiography(QWBA) and LC–MS/MS following i.v.administration of BMS-753493 or its active moiety,BMS-748285 in mice bearing FRt(98M109) and FR–(M109) tumors.QWBA showed [3H]BMS-753493–derived radioactivity was extensively distributed to various tissues.The FR over-expressing 98M109 tumors showed consistently higher level of radioactivity than FR-negative tumors(i.e.,M109 tumors) up to 48 h post dose of [3H]BMS-753493,despite the magnitude of difference between the tumors is relatively small(generally 3 5-fold).The radioactivity level in 98M109 tumors was 2 12-fold of normal tissues except intestine/content at 48 h post dose.No selective radioactivity uptake into 98M109 tumors over M109 or normal tissues was observed after i.v.administration of the active epothilone,[3H]BMS-748285.LC–MS/MS measurements demonstrated that the concentrations of BMS-748285,presumably from hydrolysis of the folate conjugate,in 98M109 tumors were greater than those in M109 tumors after i.v.administration of BMS-753493(2–3-fold) whereas no differential uptake in the tumors following BMS-748285 administration.Those data were consistent with radioactivity determinations.Those results demonstrated that the folate conjugation in BMS-753493 enabled moderately preferential distribution of the active epothilone to FR overexpressing 98M109 tumors,thereby supporting targeted delivery of cytotoxics through the folate receptor.展开更多
Objective：Human pancreatic cancer is one of the most common clinical malignancies.The effect of comprehensive treatment based on surgery is general.The effects of chemotherapy were not obvious mainly because of lack ...Objective：Human pancreatic cancer is one of the most common clinical malignancies.The effect of comprehensive treatment based on surgery is general.The effects of chemotherapy were not obvious mainly because of lack of targeting and chemoresistance in pancreatic cancer.This study aimed to investigate the effects of folate receptor （FR）-mediated gemcitabine FA-Chi-Gem nanoparticles with a core-shell structure by electrostatic spray on pancreatic cancer.Methods：In this study,the levels of expression of FR in six human pancreatic cancer cell lines were studied by immunohistochemical analysis.The uptake rate of isothiocyanate-labeled FA-Chi nanoparticles in FR high expression cell line COLO357 was assessed by fluorescence microscope and the inhibition rate of FA-Chi-Gem nanoparticles on COLO357 cells was evaluated by MTT assay.Moreover,the biodistribution of PEG-FA-ICGDER02-Chi in the orthotopic pancreatic tumor model was observed using near-infrared imaging and the human pancreatic cancer orthotopic xenografts were treated with different nanoparticles and normal saline control.Results：The expression of FR in COLO357 was the highest among the six pancreatic cancer cell lines.The FR mainly distributed on cell membrane and fewer in the cytoplasm in pancreatic cancer.Moreover,the absorption rate of the FA-Chi-Gem nanoparticles was more than the Chi nanoparticles without FA modified.The proliferation of COLO357 was significantly inhibited by FA-Chi-Gem nanoparticles.The PEG-FA-ICGDER02-Chi nanoparticles were enriched in tumor tissue in human pancreatic cancer xenografts,while non-targeted nanoparticles were mainly in normal liver tissue.PEG-FA-Gem-Chi significantly inhibited the growth of human pancreatic cancer xenografts （PEG-FA-Gem-Chi vs.Gem,t=22.950,P=0.000）.Conclusions：PEG-FA-FITC-Chi nanoparticles might be an effective targeted drug for treating human FR-positive pancreatic cancer.展开更多
Almost all cells are easily killed by exposure to potent oxidants. Indeed, major pathogen defense mechanisms in both animal and plant kingdoms involve production of an oxidative burst, where host defense cells show an...Almost all cells are easily killed by exposure to potent oxidants. Indeed, major pathogen defense mechanisms in both animal and plant kingdoms involve production of an oxidative burst, where host defense cells show an invading pathogen with reactive oxygen species (ROS). Although cancer cells can be similarly killed by ROS, development of oxidant-producing chemotherapies has been limited by their inherent nonspecificity and potential toxicity to healthy cells. In this paper, we describe the targeting of an ROS-generating molecule selectively to tumor cells using folate as the tumor-targeting ligand. For this purpose, we exploit the ability of 9,10-phenanthraquinone (PHQ) to enhance the continuous generation of H<sub>2</sub>O<sub>2</sub> in the presence of ascorbic acid to establish a con-stitutive source of ROS within the tumor mass. We report here that incubation of folate receptor-expressing KB cells in culture with folate-PHQ plus ascorbate results in the death of the cancer cells with an IC<sub>50</sub> of ~10 nM (folate-PHQ). We also demonstrate that a cleavable spacer linking folate to PHQ is significantly inferior to a noncleavable spacer, in contrast to most other folate-targeted therapeutic agents. Unfortunately, no evidence for folate-PHQ mediated tumor regression in murine tumor models is obtained, suggesting that unanticipated impediments to generation of cytotoxic quantities of ROS in vivo are encountered. Possible mechanisms and potential solutions to these unanticipated results are offered.展开更多
Background： Epidermal growth factor receptor（EGFR） mutation is the key predictor of EGFR tyrosine kinase inhibitors（TKIs） efficacy in non-small cell lung cancer（NSCLC）. We conducted this study to verify the fea...Background： Epidermal growth factor receptor（EGFR） mutation is the key predictor of EGFR tyrosine kinase inhibitors（TKIs） efficacy in non-small cell lung cancer（NSCLC）. We conducted this study to verify the feasibility of EGFR mutation analysis in cytological specimens and investigate the responsiveness to gefitinib treatment in patients carrying EGFR mutations.Methods： A total of 210 cytological specimens were collected for EGFR mutation detection by both direct sequencing and amplification refractory mutation system（ARMS）. We analyzed EGFR mutation status by both methods and evaluated the responsiveness to gefitinib treatment in patients harboring EGFR mutations by overall response rate（ORR）, disease control rate（DCR） and progression free survival（PFS）.Results： Of all patients, EGFR mutation rate was 28.6%（60/210） by direct sequencing and 45.2%（95/210） by ARMS（P〈0.001） respectively. Among the EGFR wild type patients tested by direct sequencing, 26.7% of them were positive by ARMS. For the 72 EGFR mutation positive patients treated with gefitinib, the ORR, DCR and median PFS were 69.4%, 90.2% and 9.3 months respectively. The patients whose EGFR mutation status was negative by direct sequencing but positive by ARMS had lower ORR（48.0% vs. 80.9%, P=0.004） and shorter median PFS（7.4 vs. 10.5 months, P=0.009） as compared with that of EGFR mutation positive patients by both detection methods. Conclusions： Our study verified the feasibility of EGFR analysis in cytological specimens in advanced NSCLC. ARMS is more sensitive than direct sequencing in EGFR mutation detection. EGFR Mutation status tested on cytological samples is applicable for predicting the response to gefitinib. Abundance of EGFR mutations might have an influence on TKIs efficacy.展开更多
文摘BACKGROUND Photodynamic therapy(PDT)is a minimally invasive form of cancer therapy,and the development of a novel photosensitizer(PS)with optimal properties is important for enhancing PDT efficacy.Folate receptor(FR)membrane protein is frequently overexpressed in 40%of human cancer and a good candidate for tumor-specific targeting.Specific active targeting of PS to FR can be achieved by conjugation with the folate moiety.A folate-linked,near-infrared(NIR)-sensitive probe,folate-Si-rhodamine-1(FolateSiR-1),was previously developed and is expected to be applicable to NIR-PDT.AIM To investigate the therapeutic efficacy of NIR-PDT induced by FolateSiR-1,a FRtargeted PS,in preclinical cancer models.METHODS FolateSiR-1 was developed by conjugating a folate moiety to the Si-rhodamine derivative through a negatively charged tripeptide linker.FR expression in the designated cell lines was examined by western blotting(WB).The selective binding of FolateSiR-1 to FR was confirmed in FR overexpressing KB cells(FR+)and tumors by fluorescence microscopy and in vivo fluorescence imaging.Low FR expressing OVCAR-3 and A4 cell lines were used as negative controls(FR-).The NIR light(635±3 nm)-induced phototoxic effect of FolateSiR-1 was evaluated by cell viability imaging assays.The time-dependent distribution of FolateSiR-1 and its specific accumulation in KB tumors was determined using in vivo longitudinal fluorescence imaging.The PDT effect of FolateSiR-1 was evaluated in KB tumor bearing mice divided into four experimental groups:(1)FolateSiR-1(100μmol/L)alone;(2)FolateSiR-1(100μmol/L)followed by NIR irradiation(50 J/cm2);(3)NIR irradiation(50 J/cm2)alone;and(4)no treatment.Tumor volume measurement and immunohistochemical(IHC)and histological examinations of the tumors were performed to analyze the effect of PDT.RESULTS High FR expression was observed in the KB cells by WB,but not in the OVCAR-3 and A4 cells.Substantial FR-specific binding of FolateSiR-1 was observed by in vitro and in vivo fluorescence imaging.Cell viability imaging assays showed that NIR-PDT induced cell death in KB cells.In vivo longitudinal fluorescence imaging showed rapid peak accumulation of FolateSiR-1 in the KB tumors 2 h after injection.In vivo PDT conducted at this time point caused tumor growth delay.The relative tumor volumes in the PDT group were significantly reduced compared to those in the other groups[5.81±1.74(NIR-PDT)vs 12.24±2.48(Folate-SiR-1),vs 11.84±3.67(IR),vs 12.98±2.78(Untreated),at Day 16,P<0.05].IHC analysis revealed reduced proliferation marker Ki-67-positive cells in the PDT treated tumors,and hematoxylin-eosin staining revealed features of necrotic-and apoptotic cell death.CONCLUSION FolateSiR-1 has potential for use in PDT,and FR-targeted NIR-PDT may open a new effective strategy for the treatment of FR-overexpressing tumors.
文摘Objective: To investigate the effects of linezolid and moxifloxacin combined with quadruple antituberculosis drugs on CSF cytology, NSE (neuronal specific enolase), NGF(nerve growth facor) and its receptors, endothelin and its receptors and CRP (C reaction protein) in patients with refractory tuberculous meningitis. Method: A total of 56 patients were selected with tuberculous meningitis in our hospital from February 2014 to December 2017,randomly divided them into 2 groups, each group was 28 cases, set as the observation group and the control group, both groups were treated with quadruple antituberculosis drugs, the observation group was given ilinezolid on this basis, and the control group was combined with moxifloxacin, The course of treatment was 4 weeks, compared the levels of CSF cytology, NSE, NGF and NGF receptors, endothelin and endothelin receptors, and CRP after treatment in the two groups. Result:The CSF cytology, NSE, NGF and NGF receptors, endothelin and endothelin receptor, and CRP levels remained unchanged before treatment, the difference was not statistically significant. After treatment, the chloride and glucose levels in the observation group were higher than those before treatment and that of the control group, the protein, white blood cell count, and cerebrospinal fluid pressure levels were lower than before treatment and that of the control group, the difference was statistically significant;The NSE level in the observation group after treatment was lower than before treatment and that of the control group, the difference was statistically significant;After treatment, the levels of NGF and its receptors in the observation group were higher than those before treatment and that of the control group, and the levels of endothelin, and its receptor, CRP were lower than those before treatment and that of the control group, the difference was statistically significant. Conclusion:The use of linezolid in combination with quadruple antituberculosis drugs to treat refractory tuberculous meningitis has better clinical effect, effectively improve cerebrospinal fluid cytology, regulate cerebrospinal fluid NSE levels, restore NGF, endothelin and its receptor function, reduce inflammatory response, recommended for clinical promotion and application.
基金Yunnan Provincial Department of Education Scien-tific Research Fund Project(K13225116).
文摘Autism spectrum disorder(ASD)is a serious neurodevelopmental disorder,the etiology and mechanism of which are not yet clear.Studies have shown that folate deficiency can lead to abnormalities in the de-velopment of the central nervous system.Patients with autism spectrum disorders develop folate-alpha recep-tor autoantibodies.Folate-alpha receptor autoantibodies block folate transport,leading to a deficiency of folate in nerve tissues.Folate is effective in treating patients with folate-alpha receptor autoantibodies.
基金supported by the National Natural Science Foundation of China (81072084)
文摘Invasive nonfunctional pituitary adenomas （NFPAs） are difficult to completely resect and often develop tumor recurrence after initial surgery. Currently, no medications are clinically effective in the control of NFPA. Although radiation therapy and radiosurgery are useful to prevent tumor regrowth, they are frequently withheld because of severe complications. Boron neutron capture therapy （BNCT） is a binary radiotherapy that selectively and maximally damages tumor cells without harming the surrounding normal tissue. Folate receptor （FR）-targeted boron-10 containing carbon nanoparticles is a novel boron delivery agent that can be selectively taken up by FR-expressing cells via FR-mediated endocytosis. In this study, FR-targeted boron-10 containing carbon nanoparticles were selectively taken up by NFPAs cells expressing FR but not other types of non-FR expressing pituitary adenomas. After incubation with boron-10 containing carbon nanoparticles and following irradiation with thermal neutrons, the cell viability of NFPAs was significantly decreased, while apoptotic cells were simultaneously increased. However, cells administered the same dose of FR-targeted boron-10 containing carbon nanoparticles without neutron irradiation or received the same neutron irradiation alone did not show significant decrease in cell viability or increase in apoptotic cells. The expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated in NFPAs after treatment with FR-mediated BNCT. In conclusion, FR-targeted boron-10 containing carbon nanoparticles may be an ideal delivery system of boron to NFPAs ceils for BNCT. Furthermore, our study also provides a novel insight into therapeutic strategies for invasive NFPA refractory to conventional therapy, while exploring these new applications of BNCT for tumors, especially benign tumors.
基金supported by the National Postdoctoral Program for Innovative Talents(No.BX201700142)Postdoctoral Science Foundation of China(No.2018M630732)+3 种基金the National Natural Science Foundation of China(Nos.81901805,21906135,81471707,21705037,and 91539126)Hunan Provincial Natural Science Foundation of China(No.2018JJ3092)Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(CIFMS 2016-I2M-1-009)Drug Innovation Major Project(2018ZX09711001-003-011).
文摘Vulnerable atherosclerotic plaques are responsible for most cardiovascular diseases(CVDs).Folate receptor(FR)positive activated macrophages were thought to be a prominent component in the development of vulnerable plaque.The objective of this study is to develop folate conjugated two-dimensional(2D)Pd@Au nanomaterials(Pd@Au-PEG-FA)for targeted multimodal imaging of the FRs in advanced atherosclerotic plaques.Pharmacokinetic and imaging studies(single photon emission computed tomography(SPECT),computed tomography(CT)and photoacoustic(PA)imaging)were performed to confirm the prolonged blood half-life and enrichment of radioactivity in atherosclerotic plaques.Strong signals were detected in vivo with SPECT,CT and PA imaging in heavy atherosclerotic plaques,which were significantly higher than those of the normal aortas after injection of Pd@Au-PEG-FA.Blocking studies with preinjection of excess FA could effectively reduce the targeting ability of Pd@Au-PEG-FA in atherosclerotic plaques,further demonstrating the specific binding of Pd@Au-PEG-FA for plaque lesions.Histopathological characterization revealed that the signal of probe was in accordance with the high-risk plaques.In summary,the Pd@Au-PEG-FA has favorable pharmacokinetic properties and provides a valuable approach for detecting high-risk plaques in the presence of FRs in atherosclerotic plaques.
基金supported by the National Natural Science Foundation of China (21505010, 21173274)Chongqing Research Program of Basic Research and Frontier Technology (cstc2015jcyj A1357)Scientific Research Innovation Team of Chongqing University of Technology (2015TD22)
文摘The detection of protein/small molecule interactions plays important roles in drug discovery and protein/metabolite interactions in biology. In this work, by coupling the terminal protection of small molecule-linked ss DNA strategy with the unmodified and positively charged gold nanoparticle((+)Au NP) nanoprobes, we have developed a sensitive and simple colorimetric sensor for the detection of folate receptor, a highly expressed protein in many kinds of malignant tumors. The target folate receptor binds the folate moieties of the folate-linked ss DNA through high affinity interactions and protects the protein-bound ss DNA from digestion by exonuclease I. The protected ss DNA thus adsorbs the((+)Au NP) through electrostatic interactions, leading to a red-to-blue color change of the sensing solution for sensitive colorimetric detection of folate receptor at the sub-nanomolar level. Besides, this colorimetric sensor shows high selectivity toward folate receptor against other control proteins. The developed sensor avoids the modification/conjugation of the Au NP nanoprobes and the involvement of any expensive instruments for signal transduction in protein detection. Featured with these obvious advantages, the colorimetric sensor strategy demonstrated herein can be easily expanded for sensitive and convenient detection of various protein/small molecule interactions.
文摘To assess targeting of an epothilone folate conjugate(BMS-753493) to the folate receptor(FR)-overexpressed tumor in mice bearing both FRt and FR– tumors,a series of experiments were conducted by quantitative whole-body autoradiography(QWBA) and LC–MS/MS following i.v.administration of BMS-753493 or its active moiety,BMS-748285 in mice bearing FRt(98M109) and FR–(M109) tumors.QWBA showed [3H]BMS-753493–derived radioactivity was extensively distributed to various tissues.The FR over-expressing 98M109 tumors showed consistently higher level of radioactivity than FR-negative tumors(i.e.,M109 tumors) up to 48 h post dose of [3H]BMS-753493,despite the magnitude of difference between the tumors is relatively small(generally 3 5-fold).The radioactivity level in 98M109 tumors was 2 12-fold of normal tissues except intestine/content at 48 h post dose.No selective radioactivity uptake into 98M109 tumors over M109 or normal tissues was observed after i.v.administration of the active epothilone,[3H]BMS-748285.LC–MS/MS measurements demonstrated that the concentrations of BMS-748285,presumably from hydrolysis of the folate conjugate,in 98M109 tumors were greater than those in M109 tumors after i.v.administration of BMS-753493(2–3-fold) whereas no differential uptake in the tumors following BMS-748285 administration.Those data were consistent with radioactivity determinations.Those results demonstrated that the folate conjugation in BMS-753493 enabled moderately preferential distribution of the active epothilone to FR overexpressing 98M109 tumors,thereby supporting targeted delivery of cytotoxics through the folate receptor.
基金supported by the National Natural Science Foundation of China(No.81071967 and 30872500)the Natural Science Foundation of Jiangsu province(Project No:BK2010242)
文摘Objective：Human pancreatic cancer is one of the most common clinical malignancies.The effect of comprehensive treatment based on surgery is general.The effects of chemotherapy were not obvious mainly because of lack of targeting and chemoresistance in pancreatic cancer.This study aimed to investigate the effects of folate receptor （FR）-mediated gemcitabine FA-Chi-Gem nanoparticles with a core-shell structure by electrostatic spray on pancreatic cancer.Methods：In this study,the levels of expression of FR in six human pancreatic cancer cell lines were studied by immunohistochemical analysis.The uptake rate of isothiocyanate-labeled FA-Chi nanoparticles in FR high expression cell line COLO357 was assessed by fluorescence microscope and the inhibition rate of FA-Chi-Gem nanoparticles on COLO357 cells was evaluated by MTT assay.Moreover,the biodistribution of PEG-FA-ICGDER02-Chi in the orthotopic pancreatic tumor model was observed using near-infrared imaging and the human pancreatic cancer orthotopic xenografts were treated with different nanoparticles and normal saline control.Results：The expression of FR in COLO357 was the highest among the six pancreatic cancer cell lines.The FR mainly distributed on cell membrane and fewer in the cytoplasm in pancreatic cancer.Moreover,the absorption rate of the FA-Chi-Gem nanoparticles was more than the Chi nanoparticles without FA modified.The proliferation of COLO357 was significantly inhibited by FA-Chi-Gem nanoparticles.The PEG-FA-ICGDER02-Chi nanoparticles were enriched in tumor tissue in human pancreatic cancer xenografts,while non-targeted nanoparticles were mainly in normal liver tissue.PEG-FA-Gem-Chi significantly inhibited the growth of human pancreatic cancer xenografts （PEG-FA-Gem-Chi vs.Gem,t=22.950,P=0.000）.Conclusions：PEG-FA-FITC-Chi nanoparticles might be an effective targeted drug for treating human FR-positive pancreatic cancer.
文摘Almost all cells are easily killed by exposure to potent oxidants. Indeed, major pathogen defense mechanisms in both animal and plant kingdoms involve production of an oxidative burst, where host defense cells show an invading pathogen with reactive oxygen species (ROS). Although cancer cells can be similarly killed by ROS, development of oxidant-producing chemotherapies has been limited by their inherent nonspecificity and potential toxicity to healthy cells. In this paper, we describe the targeting of an ROS-generating molecule selectively to tumor cells using folate as the tumor-targeting ligand. For this purpose, we exploit the ability of 9,10-phenanthraquinone (PHQ) to enhance the continuous generation of H<sub>2</sub>O<sub>2</sub> in the presence of ascorbic acid to establish a con-stitutive source of ROS within the tumor mass. We report here that incubation of folate receptor-expressing KB cells in culture with folate-PHQ plus ascorbate results in the death of the cancer cells with an IC<sub>50</sub> of ~10 nM (folate-PHQ). We also demonstrate that a cleavable spacer linking folate to PHQ is significantly inferior to a noncleavable spacer, in contrast to most other folate-targeted therapeutic agents. Unfortunately, no evidence for folate-PHQ mediated tumor regression in murine tumor models is obtained, suggesting that unanticipated impediments to generation of cytotoxic quantities of ROS in vivo are encountered. Possible mechanisms and potential solutions to these unanticipated results are offered.
文摘Background： Epidermal growth factor receptor（EGFR） mutation is the key predictor of EGFR tyrosine kinase inhibitors（TKIs） efficacy in non-small cell lung cancer（NSCLC）. We conducted this study to verify the feasibility of EGFR mutation analysis in cytological specimens and investigate the responsiveness to gefitinib treatment in patients carrying EGFR mutations.Methods： A total of 210 cytological specimens were collected for EGFR mutation detection by both direct sequencing and amplification refractory mutation system（ARMS）. We analyzed EGFR mutation status by both methods and evaluated the responsiveness to gefitinib treatment in patients harboring EGFR mutations by overall response rate（ORR）, disease control rate（DCR） and progression free survival（PFS）.Results： Of all patients, EGFR mutation rate was 28.6%（60/210） by direct sequencing and 45.2%（95/210） by ARMS（P〈0.001） respectively. Among the EGFR wild type patients tested by direct sequencing, 26.7% of them were positive by ARMS. For the 72 EGFR mutation positive patients treated with gefitinib, the ORR, DCR and median PFS were 69.4%, 90.2% and 9.3 months respectively. The patients whose EGFR mutation status was negative by direct sequencing but positive by ARMS had lower ORR（48.0% vs. 80.9%, P=0.004） and shorter median PFS（7.4 vs. 10.5 months, P=0.009） as compared with that of EGFR mutation positive patients by both detection methods. Conclusions： Our study verified the feasibility of EGFR analysis in cytological specimens in advanced NSCLC. ARMS is more sensitive than direct sequencing in EGFR mutation detection. EGFR Mutation status tested on cytological samples is applicable for predicting the response to gefitinib. Abundance of EGFR mutations might have an influence on TKIs efficacy.
文摘目的探究自身免疫性脑炎(autoimmune encephalitis,AE)患者的脑脊液(cerebral spinal fluid,CSF)细胞学特点。方法回顾性收集济宁医学院附属医院2016年1月至2019年6月收治的28例确诊AE患者的脑脊液常规、生化、细胞学、颅内压等结果,分析其特点及其与疾病严重程度的相关性。结果28例AE患者中,抗N-甲基-D-天冬氨酸受体(N-methyl-D-aspartate receptor,NMDAR)脑炎11例,抗富含亮氨酸胶质瘤失活蛋白1(leucine-rich glioma-inactivated protein 1,LGI1)抗体相关脑炎13例,抗γ-氨基丁酸B型受体(γ-aminobutyric acid type B receptor,GABA BR)抗体相关脑炎4例。CSF检查中,颅内压中位数(四分位数间距)为127.5(41.0)mmH 2O,蛋白中位数(四分位数间距)为0.48(0.15)g/L,糖和氯化物基本正常。28例中,7例患者CSF常规检查可见炎性反应;9例患者CSF细胞学检查可见炎性反应,其中6例为抗NMDAR脑炎患者,炎性反应类型均为淋巴细胞炎性反应,轻、中、重度炎性反应分别为4、3、2例,检测到激活淋巴细胞4例,浆细胞2例。CSF细胞学检查炎性细胞阳性率高于CSF常规检查炎性细胞阳性率(P=0.02),抗NMDAR脑炎患者CSF细胞学的炎性反应程度与患者入院时的改良Rankin评分(mRS)呈正相关(r=0.23,P<0.01)。结论本组AE患者CSF细胞学检查多表现为淋巴细胞炎性反应,抗NMDAR脑炎患者的CSF炎性反应阳性率最高,炎性反应程度最重。与CSF常规检查相比,CSF细胞学能够更敏感地发现AE的炎性反应,可作为诊断AE及评估疾病严重程度的重要方法。
