Biochimie最新文献_第3页

Enzymatic activity of HIV-1 protease defines migration of tumor cells in vitro and enhances their metastatic activity in vivo. hiv-1 蛋白酶的酶活性决定了肿瘤细胞在体外的迁移并增强其在体内的转移活性。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-08-14 DOI: 10.1016/j.biochi.2024.08.009

M Isaguliants, A Zhitkevich, S Petkov, T Gorodnicheva, D Mezale, I Fridrihsone, Y Kuzmenko, D Kostyushev, A Kostyusheva, I Gordeychuk, E Bayurova

{"title":"Enzymatic activity of HIV-1 protease defines migration of tumor cells in vitro and enhances their metastatic activity in vivo.","authors":"M Isaguliants, A Zhitkevich, S Petkov, T Gorodnicheva, D Mezale, I Fridrihsone, Y Kuzmenko, D Kostyushev, A Kostyusheva, I Gordeychuk, E Bayurova","doi":"10.1016/j.biochi.2024.08.009","DOIUrl":"10.1016/j.biochi.2024.08.009","url":null,"abstract":"Overexpression of aspartic proteases, as cathepsin D, is an independent marker of poor prognosis in breast cancer, correlated with the incidence of clinical metastasis. We aimed to find if HIV-1 aspartic protease (PR) can play a similar role. Murine adenocarcinoma 4T1luc2 cells were transduced with lentivirus encoding inactivated drug-resistant PR, generating subclones PR20.1 and PR20.2. Subclones were assessed for production of reactive oxygen species (ROS), expression of epithelial-mesenchymal transition (EMT) factors, and in vitro migratory activity in the presence or absence of antioxidant N-acetyl cysteine and protease inhibitors. Tumorigenic activity was evaluated by implanting cells into BALB/c mice and following tumor growth by calipering and bioluminescence imaging in vivo, and metastases, by organ imaging ex vivo. Both subclones expressed PR mRNA, and PR20.2, also the protein detected by Western blotting. PR did not induce production of ROS, and had no direct effect on cell migration rate, however, treatment with inhibitors of drug-resistant PR suppressed the migratory activity of both subclones. Furthermore, expression of N-cadherin and Vimentin in PR20.2 cells and their migration were enhanced by antioxidant treatment. Sensitivity of in vitro migration to protease inhibitors and to antioxidant, known to restore PR activity, related the effects to the enzymatic activity of PR. In vivo, PR20.2 cells demonstrated higher tumorigenic and metastatic activity than PR20.1 or parental cells. Thus, HIV-1 protease expressed in breast cancer cells determines their migration in vitro and metastatic activity in vivo. This effect may aggravate clinical course of cancers in people living with HIV-1.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"32-43"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141918326","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

Using targeted proteomics-based detection of collagen propeptides to quantify fibrillar collagen biogenesis in vitro. 利用基于蛋白质组学的胶原蛋白肽靶向检测，量化体外纤维胶原蛋白的生物生成。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-09-14 DOI: 10.1016/j.biochi.2024.09.004

Matthias Kühle, Joachim Kuhn, Thanh-Diep Ly, Cornelius Knabbe, Bastian Fischer

{"title":"Using targeted proteomics-based detection of collagen propeptides to quantify fibrillar collagen biogenesis in vitro.","authors":"Matthias Kühle, Joachim Kuhn, Thanh-Diep Ly, Cornelius Knabbe, Bastian Fischer","doi":"10.1016/j.biochi.2024.09.004","DOIUrl":"10.1016/j.biochi.2024.09.004","url":null,"abstract":"The collagen superfamily, as the major structural component of the extracellular matrix, encompasses 28 distinct subtypes, with type-I and -III forming fibrils crucial for the matrix scaffold. During collagen biogenesis, trimers of type-I and -III procollagen are secreted into the extracellular matrix. The N- and C-terminal propeptides of these trimers are proteolytically cleaved from procollagen during secretion, initiating collagen fibril formation. The propeptides are released into extracellular space and, therefore, have been used to quantify collagen biogenesis. But high-throughput methods for the quantification of these biomarkers are still lacking. This study presents a state-of-the-art multiplexed approach for the simultaneous quantification of PINP, PICP, PIIINP and PIIICP from cell culture supernatants. The ability of targeted proteomics to quantify these propeptides from cell culture samples was assessed in this study. Using tryptic digestion and solid phase extraction, we were able to accurately quantify precollagen propeptides in a range of 3-1000 ng/mL. The assay showed an average inter-assay variance of 6.86 % with an overall recovery ranging from 92 to 98 %. The assay was validated using recombinant protein standards diluted in surrogate matrix and tested using transforming growth factor β1 mediated induction of normal human dermal fibroblasts. In summary, the assay presented in this paper offers a novel, robust, and precise high-throughput method for measuring human collagen propeptides in cell culture supernatants, empowering researchers to assess collagen biogenesis effectively in in vitro experiments.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"101-113"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142303199","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

Extracellular vesicles as a potential source of biomarkers for endocrine disruptors in MASLD: A short review on the case of DEHP. 细胞外囊泡作为 MASLD 中内分泌干扰物生物标志物的潜在来源：DEHP 案例简评。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-09-21 DOI: 10.1016/j.biochi.2024.09.009

Pierre-Etienne Merret, Lydie Sparfel, Catherine Lavau, Dominique Lagadic-Gossmann, Corinne Martin-Chouly

引用次数: 0

Mutations in the N-domain of aryl hydrocarbon receptor interacting protein affect interactions with heat shock protein 90β and phosphodiesterase 4A5. 芳基烃受体相互作用蛋白 N-域的突变会影响与热休克蛋白 90β 和磷酸二酯酶 4A5 的相互作用。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-09-18 DOI: 10.1016/j.biochi.2024.09.005

Marita Vella, Iain W Manfield, Brandon C Seychell, Chi H Trinh, Robert Rambo, G Nasir Khan, Josanne Vassallo, Thérèse Hunter, Gary J Hunter

{"title":"Mutations in the N-domain of aryl hydrocarbon receptor interacting protein affect interactions with heat shock protein 90β and phosphodiesterase 4A5.","authors":"Marita Vella, Iain W Manfield, Brandon C Seychell, Chi H Trinh, Robert Rambo, G Nasir Khan, Josanne Vassallo, Thérèse Hunter, Gary J Hunter","doi":"10.1016/j.biochi.2024.09.005","DOIUrl":"10.1016/j.biochi.2024.09.005","url":null,"abstract":"The aryl hydrocarbon receptor interacting protein (AIP) is a cytoplasmic molecular co-chaperone and tumour suppressor that assists in protein stability and complex formation involving the aryl hydrocarbon receptor. Germline mutations in the AIP gene predispose to pituitary tumourigenesis with patients exhibiting an aggressive clinical phenotype. Full length AIP proteins harbouring N-domain mutations (R9Q, R16H, V49 M and K103R) were purified from E.coli utilizing a methodology that maintained structural integrity and monomeric stability. Mutations did not significantly affect the thermal stability of the protein and caused no overall disruptive effect in the protein structure. The mutations studied lowered the binding affinity of AIP towards two of its binding partners; heat shock protein 90β and phosphodiesterase 4A5 (PDE4A5). The inhibition of phosphodiesterase activity by AIP was also greatly reduced by all mutants. While previously published data has mainly concentrated on the tetratricopeptide repeats of the C-domain of AIP, we present clear evidence that AIP N-domain mutations play a significant role in two protein:protein interactions with partner proteins. The complex interactome of AIP suggests that any observable change in one or more of its binding partners cannot be disregarded as it may have repercussions on other biochemical pathways.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"114-126"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142303296","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

The Barnase-Barstar-based pre-targeting strategy for enhanced antitumor therapy in vivo. 基于 Barnase-Barstar 的预靶向策略用于增强体内抗肿瘤治疗。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-09-21 DOI: 10.1016/j.biochi.2024.09.011

G M Proshkina, E I Shramova, A B Mirkasyimov, O Yu Griaznova, E V Konovalova, A A Schulga, S M Deyev

{"title":"The Barnase-Barstar-based pre-targeting strategy for enhanced antitumor therapy in vivo.","authors":"G M Proshkina, E I Shramova, A B Mirkasyimov, O Yu Griaznova, E V Konovalova, A A Schulga, S M Deyev","doi":"10.1016/j.biochi.2024.09.011","DOIUrl":"10.1016/j.biochi.2024.09.011","url":null,"abstract":"There is a great need for novel approaches to the treatment of epithelial ovarian carcinoma, which is the leading cause of mortality from gynecological malignancies. In this study, the pre-targeting technology was used to enhance the in vivo targeting of cytotoxic module composed of nanoliposomes loaded with a truncated form of Pseudomonas aeruginosa exotoxin A (PE40) to cancer cells. Pre-targeting system used in this study is composed of bacterial ribonuclease Barnase and its natural antitoxin Barstar. Barstar, genetically fused to various engineered scaffold proteins specific to tumor-associated antigens (HER2, EpCAM) serves as a primary module for precise cancer cell recognition. Barnase conjugated to a therapeutic agent serves as a cytotoxic or secondary module for malignant cell elimination. Due to strong non-covalent interaction (KD10-14 M) of Barstar and Barnase, the primary and secondary modules efficiently interact with each other on the cell surface, which has been proven by confocal microscopy and flow cytometry. Using mice with SKOV-3 ovarian cancer xenografts, we have shown that regardless of the targeting module, the pre-targeting approach is much more effective than a single-step active targeting.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"158-166"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142303198","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

Developmental overlap between skeletal muscle maturation and perirenal fat brown-to-white transition in goats: Exploring the role of Myf-5. 山羊骨骼肌成熟与肾周脂肪褐色到白色转变之间的发育重叠：探索 Myf-5 的作用。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-08-07 DOI: 10.1016/j.biochi.2024.08.005

Sunil Pani, Unmod Senapati, Bijayashree Sahu, Benudhara Pati, Gourabamani Swalsingh, Punyadhara Pani, Birendra Kumar Bindhani, K Gopinath Achary, Naresh C Bal

{"title":"Developmental overlap between skeletal muscle maturation and perirenal fat brown-to-white transition in goats: Exploring the role of Myf-5.","authors":"Sunil Pani, Unmod Senapati, Bijayashree Sahu, Benudhara Pati, Gourabamani Swalsingh, Punyadhara Pani, Birendra Kumar Bindhani, K Gopinath Achary, Naresh C Bal","doi":"10.1016/j.biochi.2024.08.005","DOIUrl":"10.1016/j.biochi.2024.08.005","url":null,"abstract":"In mammals, skeletal muscles (SkMs) and adipose tissues regulate energy homeostasis and share developmental origins. Notably, the perirenal adipose tissue (PRAT) depot has been reported to display adipocyte heterogeneity: while some originated from Myogenic factor 5 (Myf-5) expressing progenitors, others did not. Our study examines the expression and distribution of Myf-5 using immunohistochemical staining and western blotting of PRAT, gastrocnemius, and trapezius from goat at various developmental stages. Contrary to earlier beliefs, functionally divergent SkM gastrocnemius and trapezius showed similar Myf-5 expressional pattern. SkM abundantly expresses Myf-5 in developing myocytes which gradually becomes limited to the nucleus of myogenic stem cells and is retained only in a few differentiated postnatal fibers. During the same period, PRAT displays a unique brown-to-white transition. PRAT exhibited an elevated expression of Myf-5 during prenatal periods, which declines thereafter and becomes negligible during adulthood where it gets fully enriched white adipocytes. The reduction of Myf-5 during the neonatal period was common to all three tissues. However, Myf-5 expression was retained in some of the differentiated myofibers while it was undetectable in adult PRAT. These observations suggest a possible developmental interplay between adipose tissue and SkM where Myf-5 might be a major regulator.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141914858","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

New transaminase from Odontosyllis undecimdonta the first potential enzyme of the luciferin biosynthesis pathway. 来自 Odontosyllis undecimdonta 的新转氨酶是荧光素生物合成途径的第一个潜在酶。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-08-31 DOI: 10.1016/j.biochi.2024.08.012

Daria A Dmitrieva, Olga A Belozerova, Alexey V Mishin, Ilia V Yampolsky, Alexey A Kotlobay

引用次数: 0

Unveiling the mechanism of hesperidin-induced LdTopI-mediated cell death pathway in protozoan parasite Leishmania donovani. 揭示原生动物寄生虫唐氏利什曼原虫中橙皮甙诱导 LdTopI 介导的细胞死亡途径的机制。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-08-10 DOI: 10.1016/j.biochi.2024.08.008

Swati Lamba, Priyanka Mazire, Amit Roy

{"title":"Unveiling the mechanism of hesperidin-induced LdTopI-mediated cell death pathway in protozoan parasite Leishmania donovani.","authors":"Swati Lamba, Priyanka Mazire, Amit Roy","doi":"10.1016/j.biochi.2024.08.008","DOIUrl":"10.1016/j.biochi.2024.08.008","url":null,"abstract":"Unicellular protozoan parasite Leishmania donovani is the causative agent for visceral leishmaniasis (VL) or Kala-azar, a neglected fatal parasitic disease. The conventional treatment of VL consists of therapeutic agents having several shortcomings such as toxicity, high cost, efficacy variance and increased drug resistance. Therefore, there is a desperate need to develop an effective treatment against the parasite. Previous reports suggested that flavonoids can inhibit the enzyme Leishmania donovani DNA topoisomerase I (LdTopILS). Therefore, for the first time in this present study, we divulge HSP (one of the natural sources of flavonoids), as a potent natural antileishmanial compound with efficacy in BALB/c mice at 20 mg/kg of body weight, inhibits LdTopILS at 97 % of its activity at 160 μM in preincubation condition (competitively). It binds with free enzyme and does not allow it to bind with the substrate DNA. Moreover, HSP does not stabilize DNA-topoisomerase I cleavable complex. Thus, HSP acts a catalytic topoisomerase I inhibitor, which inhibits complete activity by binding with Lys269 and Thr411 of large subunit of the enzyme. On the other hand, HSP induces the topo I-mediated programmed cell death process by the formation of cellular reactive oxygen species, resulting in depolarization of mitochondrial membrane potential, followed by fragmentation of nuclear DNA. Therefore, the present study illuminates a natural flavonoid that in future might be a promising lead for the treatment of VL.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"15-31"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141918365","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

Fibrinolytic enzyme from Arthrospira platensis and its effects on breast cancer cells: Exploring its potential as an innovative therapy. 来自铂氏螺旋体的纤溶酶及其对乳腺癌细胞的作用：探索其作为一种创新疗法的潜力。

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.12.013

Yanara Alessandra Santana Moura, Marllyn Marques da Silva, Sara Cadete da Silva, Thiago Pajeú Nascimento, Ana Cristina Lima Leite, Milena Tereza Torres do Couto, Mariane Cajubá de Britto Lira-Nogueira, Tamiris Alves Rocha, Ana Lúcia Figueiredo Porto, Raquel Pedrosa Bezerra

{"title":"Fibrinolytic enzyme from Arthrospira platensis and its effects on breast cancer cells: Exploring its potential as an innovative therapy.","authors":"Yanara Alessandra Santana Moura, Marllyn Marques da Silva, Sara Cadete da Silva, Thiago Pajeú Nascimento, Ana Cristina Lima Leite, Milena Tereza Torres do Couto, Mariane Cajubá de Britto Lira-Nogueira, Tamiris Alves Rocha, Ana Lúcia Figueiredo Porto, Raquel Pedrosa Bezerra","doi":"10.1016/j.biochi.2024.12.013","DOIUrl":"10.1016/j.biochi.2024.12.013","url":null,"abstract":"Fibrinolytic enzymes are promising in treating cardiovascular diseases due to their capacity to dissolve blood clots. The fibrinolytic enzyme from Arthrospira platensis (FEAP) was purified by ion exchange chromatography to investigate its ability to activate plasminogen, as well as its thrombolytic and fibrinogenolytic potential. Subsequently, two different cytotoxic assays (MTT and NR) and hemolysis test were performed to evaluate FEAP's safety. Furthermore, cell migration and the genotoxic and hemolytic potential were also investigated. The purified enzyme showed thrombus degradation of 43 % and thrombolytic action directly on fibrin, which can reduce possible side effects, such as hemorrhage. MTT assay was more sensitive to determine the enzyme cytotoxicity, which decreased the viability of breast cancer tumor cells (Sarcoma-180 and MDA-MB-231) and macrophages (J774A.1). In addition, the enzyme also exhibited non-hemolytic, antimetastatic, and non-genotoxic characteristics. These findings are innovative for a fibrinolytic protease and may indicate that it is safe for people undergoing cancer treatment, reducing side effects such as hemorrhage, in addition to inhibiting tumor cells and preventing metastasis, which can help with chemotherapy treatment.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142924185","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

Enhanced activity of split trehalase biosensors by interspecies domain combineering. 通过种间结构域组合提高分离式三卤甲烷酶生物传感器的活性。

Biochimie Pub Date : 2025-01-01 Epub Date: 2024-11-04 DOI: 10.1016/j.biochi.2024.09.013

Yongpeng Fu, Jeroen De Buck

{"title":"Enhanced activity of split trehalase biosensors by interspecies domain combineering.","authors":"Yongpeng Fu, Jeroen De Buck","doi":"10.1016/j.biochi.2024.09.013","DOIUrl":"10.1016/j.biochi.2024.09.013","url":null,"abstract":"The split trehalase biosensor has potential as a versatile diagnostic technology. Split enzymes are engineered proteins, divided into inactive fragments, which can reassemble and regain activity when brought together by an analyte. The split TreA biosensor requires no sample processing and produces stable signals (in the form of glucose). Split trehalase reagents can function in blood, but periplasmic trehalase of E. coli requires blood acidification for maximal activity. The objective of this study was to obtain split trehalase with near physiological pH optimum. For this purpose, periplasmic trehalases of Cellvibrio spp. with higher activity at neutral pH, were split in analogy with the E. coli TreA into hood and catalytic domains. However, these split trehalases displayed self-complementation due to spontaneous reassembly. In contrast, when catalytic domains of Cellvibrio trehalases were combined with E. coli hood domains, these hybrids displayed conditional complementation capacity when split trehalase fragments fused to immunoglobulin-binding protein G (STIGA) were used to quantify immunoglobulin concentrations. Other hybrid combinations of Cellvibrio spp. had increased activity compared to the cognate pairs, albeit with strong self-complementation. A mutagenesis analysis of residues responsible for self-complementation led to uncoupling of self-complementation from allostery. The Michaelis-Menten kinetics of Cellvibrio enzymes and fragment pairs confirmed improved activity of a mutated hybrid pair of Cellvibrio hood and catalytic domains at physiological pH. In conclusion, the improvements in pH optimum and activity, demonstrated with STIGA, can now be leveraged to enhance other variations of the split trehalase biosensor platform, broadening its utility for testing clinical samples.","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":" ","pages":"167-175"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142585151","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