Bioscience Horizons最新文献

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Orexin and melanin-concentrating hormone neurons: a hypothalamic interface for sleep and feeding regulation 食欲素和黑色素集中激素神经元:睡眠和进食调节的下丘脑界面
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU008
L. O’Leary
{"title":"Orexin and melanin-concentrating hormone neurons: a hypothalamic interface for sleep and feeding regulation","authors":"L. O’Leary","doi":"10.1093/BIOHORIZONS/HZU008","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU008","url":null,"abstract":"© The Author 2014. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. 1 Orexin and melanin-concentrating hormone neurons: a hypothalamic interface for sleep and feeding regulation","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Stem cells slow cognitive decline in Alzheimer's disease via neurotrophin action 干细胞通过神经营养素的作用减缓阿尔茨海默病的认知能力下降
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU013
R. Mulvey
{"title":"Stem cells slow cognitive decline in Alzheimer's disease via neurotrophin action","authors":"R. Mulvey","doi":"10.1093/BIOHORIZONS/HZU013","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU013","url":null,"abstract":"Alzheimer’s disease is a growing concern with no satisfactory current treatment solution. Contemporary stem cell research offers a new arena for development in this field. Transplantation of stem cells into the damaged brain brings hope of repair to damaged neurons. This appears to operate via a ‘bystander effect’ whereby neurotrophins secreted by the cells act as a neuro protectant, rather than a cell replacement mechanism as some have postulated. Such treatments can slow or even reverse cognitive decline. Research into neural stem cell transplantation has shown reversal of cognitive decline in animal models of disease via the mechanism of brain-derived neurotrophic factor secretion. Studies using nerve growth factor secreting stem cells have showed promising results with cognitive decline reversed in animal models of the disease. A Phase 1 clinical trial also showed promising reversal of cognitive decline in human subjects using transplantation of nerve growth factor secreting fibroblasts. Mesenchymal stem cells have also shown promise, and results from human trials are awaited. Induced pluripotent stem cells have provided a successful model of human disease in vitro. Although early results from transplant studies are encouraging, a lot more research will be needed before these preliminary advances can be translated to therapies with a strong evidence base to be used in practice.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Cell therapy for multiple sclerosis: a new hope 多发性硬化症的细胞疗法:新的希望
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU014
S. Harbison
{"title":"Cell therapy for multiple sclerosis: a new hope","authors":"S. Harbison","doi":"10.1093/BIOHORIZONS/HZU014","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU014","url":null,"abstract":"Multiple sclerosis is a chronic demyelinating autoimmune disease with uncertain aetiology. Due to the heterogeneity of the disease, patients may present with a wide variety of neurological symptoms such as optic neuritis, sensory deficits or cerebel lar dysfunction. It remains a disease showing little hope in terms of finding a cure. Although current therapies, such as interferon-β and glatiramer acetate, provide symptomatic relief and can delay the degenerative process, there is still a large impact on quality of life as these therapies lack an ability to reverse damage occurring prior to treatment. Recently, cell therapy has emerged as a promising treatment with signs of recovery both pathologically and clinically in a variety of animal models. Given the multifaceted capabilities of the various stem cells, including immunomodulation and neuroprotection, their potential use as a comprehensive therapy is much more promising than any pharmacological therapy to date. Here, the latest advances of cell therapy are discussed, in terms of potential efficacy, the various cell types that are used, their mecha nisms of action and the obstacles that still need to be overcome for translation into a clinical setting.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765334","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Effects of superoxide donor menadione in adult Rat myocardium are associated with increased diastolic intracellular calcium 超氧供体甲萘醌对成年大鼠心肌的影响与舒张期细胞内钙升高有关
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZT012
L. Rogers, A. Lake, K. White, M. Hardy, E. White
{"title":"Effects of superoxide donor menadione in adult Rat myocardium are associated with increased diastolic intracellular calcium","authors":"L. Rogers, A. Lake, K. White, M. Hardy, E. White","doi":"10.1093/BIOHORIZONS/HZT012","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZT012","url":null,"abstract":"Superoxide anions have been associated with many aspects of cardiovascular disease. Menadione is a superoxide anion donor that alters the heart’s electrical and mechanical functions. The aim of this study was to demonstrate simultaneous changes in intracellular Ca 2+ ([Ca 2+ ]i) and mechanical activity in intact adult cardiac myocytes, and mechanical activity and electrical activity in isolated whole hearts in order to provide greater insight into the mechanisms associated with the detrimental effects of menadione on the myocardium. Isolated hearts from adult male Wistar rats (n = 11, 200–250 g) were Langendorff perfused at 38°C with a Krebs–Henseleit solution. A saline-filled balloon was placed in the left ventricle (LV) in order to measure diastolic and developed pressure. Monophasic action potentials were simultaneously recorded from the epicardial surface. External stimulation at 5 Hz and intrinsic pacing were used throughout a 10 min control period and 30 min exposure to 50 µM menadione. Single LV myocytes (n = 7 from n = 4 animals) were loaded with the Ca2+-indicator Fura4-AM, stimulated at 1 Hz and exposed to 50 µM menadione. Myocyte length was simultaneously measured with [Ca 2+ ]i using a video edge detection system. In isolated hearts, exposure to menadione significantly decreased contractility and action potential duration (with a simi lar time course); intrinsic heart rate and rhythmicity. Diastolic pressure was significantly increased. In single adult myocytes, menadione caused a significant increase in diastolic [Ca 2+ ]i and a decrease in resting cell length and led to spontaneous release of [Ca 2+ ]i. We conclude that the effects of menadione upon electrical and mechanical activity of the heart are at least in part a consequence of dysregulation of [Ca 2+ ]i handling and the subsequent increase in diastolic [Ca 2+ ] alterations in [Ca 2+ ]i are consistent with the generation of delayed after depolarization arrhythmias.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZT012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
One step forward, two steps back: the evolution of phytoremediation into commercial technologies 前进一步,后退两步:植物修复技术向商业技术的演变
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU009
C. Stephenson, C. Black
{"title":"One step forward, two steps back: the evolution of phytoremediation into commercial technologies","authors":"C. Stephenson, C. Black","doi":"10.1093/BIOHORIZONS/HZU009","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU009","url":null,"abstract":"This review charts the evolution of phytoremediation from its earliest beginnings, with the discovery of metal tolerant plants in the 16th century and metabolism of organic pollutants by plants in the 1940s. The rapid expansion of research in the early 1990s led to many crucial discoveries but failed to surmount the fundamental limitations that often impede commercial application of phytoremediation. It is argued that phytoremediation was saved from being forgotten by its evolution under the new term phytotechnology, or ‘the application of science and engineering to examine problems and provide solutions using plants’. This review explores the use of phytotechnology for ecological engineering using constructed wetlands and evapotranspiration caps as landfill covers. Finally, the transfer of phytotechnology to developing countries, where it has great potential to solve the growing problem of pollution, is examined. The development of phytotechnology can be perceived as an illustration of the modern evolution of scientific thought, from the traditional reductionist view to a wider holistic approach which takes into account the natural environment and our need to preserve it. It is hoped that the evolution of both will allow for increasing conservation of finite resources without sacrificing continued development.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 49
Development of an anti-CD2/CD3/CD28 bead-based T-cell proliferation assay 抗cd2 /CD3/CD28头t细胞增殖试验的建立
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU012
L. Green
{"title":"Development of an anti-CD2/CD3/CD28 bead-based T-cell proliferation assay","authors":"L. Green","doi":"10.1093/BIOHORIZONS/HZU012","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU012","url":null,"abstract":"© The Author 2014. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. 1 Development of an anti-CD2/CD3/CD28 bead-based T-cell proliferation assay","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 7
The synergistic effect of gentamicin and ceftazidime against Pseudomonas fluorescens 庆大霉素与头孢他啶对荧光假单胞菌的协同作用
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU007
A. Morgan
{"title":"The synergistic effect of gentamicin and ceftazidime against Pseudomonas fluorescens","authors":"A. Morgan","doi":"10.1093/BIOHORIZONS/HZU007","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU007","url":null,"abstract":"along with the fractional inhibitory concentration index (FICI) to test for synergy. A value of ≤0.5 defined synergy; 0.5 < FICI < 4 defined no interaction; ≥4 defined antagonism. No results of synergy were found; there were five results of no interaction and six results of antagonism. The MIC of ceftazidime was 3 µg/ml and the MBC was 4 µg/ml. The MIC of gentamicin was 0.25 µg/ ml and the MBC was 3 µg/ml. The combination of gentamicin and ceftazidime is optimal at a volume ratio of 1:1, in this case 25 µl gentamicin/25 µl ceftazidime, where gentamicin has a concentration of 0.5 µg/ml and ceftazidime has a concentration of 0.25 µg/ml, when used against 50 µl of 1–2 × 10 6 colony forming units per millilitre of P. fluorescens in vitro. This study recommends that this combination therapy be studied in depth in vivo, and that clinicians understand that this combination of antibiotics does not have a synergistic effect when treating patients in this manner.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 11
Testing the disturbance hiss of the Madagascar hissing Cockroach (Gromphadorhina portentosa) as an anti-predatory response 马达加斯加嘶嘶蜚蠊(Gromphadorhina portentosa)的干扰嘶嘶反掠食性反应试验
Bioscience Horizons Pub Date : 2014-01-01 DOI: 10.1093/BIOHORIZONS/HZU010
R. Shotton
{"title":"Testing the disturbance hiss of the Madagascar hissing Cockroach (Gromphadorhina portentosa) as an anti-predatory response","authors":"R. Shotton","doi":"10.1093/BIOHORIZONS/HZU010","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZU010","url":null,"abstract":"The display of the disturbance hiss by the Madagascar Hissing Cockroach ( Gromphadorhina portentosa ) is considered to be an anti-predatory response despite there being little direct evidence linking the hiss with survival. Many studies have investi-gated the roles of the aggression and courtship hisses, displayed in this social species, but few have considered the role of the disturbance hiss. This study looked at the stimulus for the disturbance hiss response by placing unfamiliar individuals into different social contexts. A total of 10 male and 10 female G. portentosa were kept separately before being placed into four different social situations for 5 min at a time. The individuals were placed into an established colony of all females , an established colony of all males and an established colony of mixed sex G. portentosa . The subjects were also placed in the presence of a predator, an Emperor Scorpion ( Pandinus imperator Koch). All interactions and hisses were recorded both by video and on a sound-recording device. There was a highly significant difference in the setting in which the disturbance hiss was shown and a highly significant difference in the display rates of the disturbance hiss between the sexes in general, with most displays of the disturbance hiss being when introduced to a mixed sex colony. The findings suggest that the role of the disturbance hiss is not an anti-predatory response when presented with a predator of limited auditory senses. Further study into the behavioural ecology of this species is recommended to understand the range of anti-predatory responses used by this species when presented with different predators. It was also found that there is some social context for the display of the disturbance hiss which warrants further study.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"24 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZU010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60765724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
A scientific review: mechanisms of valproate-mediated teratogenesis 科学综述:丙戊酸介导的致畸机制
Bioscience Horizons Pub Date : 2013-01-01 DOI: 10.1093/BIOHORIZONS/HZT003
K. Lloyd
{"title":"A scientific review: mechanisms of valproate-mediated teratogenesis","authors":"K. Lloyd","doi":"10.1093/BIOHORIZONS/HZT003","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZT003","url":null,"abstract":"Living with epilepsy, for the majority of patients, is a continuous cycle of careful monitoring and attention to health. When coupled with pregnancy, safety becomes the primary focus. Epileptic women with reproductive intentions are confronted with a dilemma to either continue or discontinue current medication. Remaining on medication incurs the possibility of foetal abnormalities. However, discontinuation of medication could result in an uncontrolled condition that is potentially life threatening for both mother and child. Valproic acid (VPA) is a monocarboxylic acid with unclear mechanism of action and this is widely prescribed for epilepsy, bipolar disorder, migraine, Alzheimer’s disease and recently, cancer and HIV polytherapies. Originally, low incidences of toxicity were reported; however, emerging teratogenic properties warranted further investigation. This paper examines the major contraindications that VPA facilitates during pregnancy and proposes the degree of teratogenic influence each may inflict upon the developing foetus. The relevant aspects addressed are drug accumulation within the foetus, oxidative stress, folate antagonism and histone deacetylase (HDAC) inhibition. Review of the literature has shown vast numbers of investigations proving and disproving various proposed mechanisms of VPA-induced teratogenicity; these are still largely undefined. Based on the evidence presented, HDAC inhibition provided the strongest association with terato genicity, followed closely by the formation of reactive oxygen species. Both were particularly influential in the first trimester of pregnancy when DNA dysregulation has the largest impact on foetal organ development. In comparison, drug accumulation posed a lower risk as VPA is not the primary substrate for the majority of drug transporters across the placenta and lastly, folate inhibition was considered the lowest risk as new evidence has highlighted that the natural progression of gestation increases folate deficiency irrespective of VPA. All the suggested mechanisms (and possibly many more) may be contributing factors, together with inter-patient variability, environmental and lifestyle factors, each of which is also undefined, and lead to an increased risk of the teratogenic effects of VPA.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZT003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60764860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 47
Preliminary investigation of the effects of silencing the non-coding RNA, NEAT1, on the Burkitt's lymphoma cell line BJAB 沉默非编码RNA NEAT1对伯基特淋巴瘤细胞系BJAB影响的初步研究
Bioscience Horizons Pub Date : 2013-01-01 DOI: 10.1093/BIOHORIZONS/HZT006
C. Halford
{"title":"Preliminary investigation of the effects of silencing the non-coding RNA, NEAT1, on the Burkitt's lymphoma cell line BJAB","authors":"C. Halford","doi":"10.1093/BIOHORIZONS/HZT006","DOIUrl":"https://doi.org/10.1093/BIOHORIZONS/HZT006","url":null,"abstract":"Nuclear enriched abundant transcript 1 (NEAT1) is a long non-coding RNA with two isoforms. Both are expressed constitutively and have been shown to play a crucial structural role in the formation of paraspeckles. Paraspeckles are subnuclear bodies which retain certain mRNAs, preventing their translation in the cytoplasm, thereby silencing the corresponding genes. This study aimed to assess the effects of knocking down NEAT1 by RNA interference on the Burkitt’s lymphoma cell line, BJAB. The results have shown that a targeted siRNA depletion of NEAT1 resulted in an increased number of cells displaying aberrant morphology, including the appearance of ‘giant cells’, multinucleated cells and cells with cytoplasmic vacuoles. Furthermore, NEAT1 down-regulation was accompanied with an increased level of apoptosis and decreased cell viability, assessed by acridine orange morphology staining, vital dye exclusion and (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4sulfophenyl)-2H-tetrazolium) assay. Taken together, the study strongly suggests that modulation of NEAT1 expression has a significant effect on the BJAB cell line. Such remarkable effects of changes of NEAT1 expression on the control of cell morphol ogy reported here indicate the potential significance of this gene in the development and progression of cancer and highlight the importance of further investigation into the role of NEAT1 dysregulation in autoimmune disease and oncogenesis.","PeriodicalId":52095,"journal":{"name":"Bioscience Horizons","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/BIOHORIZONS/HZT006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60764958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
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