The Protein Journal最新文献_第8页

Comparative Characterization of Plasmodium falciparum Small Heat Shock Proteins and Their Inhibition by Quercetin (3,3′,4′,5,7-Pentahydroxyflavone) 恶性疟原虫小热休克蛋白的比较鉴定及其槲皮素（3,3',4',5,7-五羟基黄酮）的抑制作用。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-07-18 DOI: 10.1007/s10930-025-10281-w

Francisca Magum Timothy, Tawanda Zininga

{"title":"Comparative Characterization of Plasmodium falciparum Small Heat Shock Proteins and Their Inhibition by Quercetin (3,3′,4′,5,7-Pentahydroxyflavone)","authors":"Francisca Magum Timothy, Tawanda Zininga","doi":"10.1007/s10930-025-10281-w","DOIUrl":"10.1007/s10930-025-10281-w","url":null,"abstract":"<div><p><i>Plasmodium falciparum</i>, the parasite responsible for the most severe form of malaria, encodes three small heat shock proteins (sHsps), PfHsp20a, PfHsp20b, and PfHsp20c, each containing a conserved α-crystallin domain (ACD). These class I sHsps are hypothesized to play critical roles in proteostasis under stress, yet their specific functions have remained poorly defined. In this study, all three sHsps were recombinantly expressed and purified for structural and functional characterization. Circular dichroism and thermal shift assays revealed distinct conformational properties, with PfHsp20a exhibiting the highest thermal and chemical stability. Functional assays using malate dehydrogenase and citrate synthase confirmed that all three isoforms possess autonomous chaperone activity, although with varying efficiency. Notably, the plant-derived flavonoid quercetin disrupted both the structure and function of the sHsps in a concentration-dependent manner, with PfHsp20c being the most sensitive. Quercetin also inhibited the growth of <i>P. falciparum</i> Nf54 and Dd2 strains in vitro with IC<sub>50</sub> values of 5.4 μM and 7.8 μM, respectively. These results provide the first direct evidence of independent chaperone activity in <i>P. falciparum</i> sHsps and highlight their vulnerability to small molecule inhibition. This establishes their potential as novel drug targets for antimalarial intervention.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 5","pages":"580 - 597"},"PeriodicalIF":1.4,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s10930-025-10281-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144669245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Functional Activities of Cohesin Proteins Can Be Altered by Chemical Chaperones 黏结蛋白的功能活性可被化学伴侣改变。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-07-16 DOI: 10.1007/s10930-025-10276-7

Sayali Marathe, Haripriya Chougule, Vandana Nikam, Amitabha Majumdar, Tania Bose

引用次数: 0

Mechanism-Based Allosteric Inhibition of PTP1B by Prenylated Flavonoids from Glycyrrhiza echinata: In Vitro Experiments and in Silico Validation 基于机制的甘草烯酰化黄酮类化合物对PTP1B的变构抑制：体外实验和硅验证。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-07-16 DOI: 10.1007/s10930-025-10272-x

Emadeldin M. Kamel, Doaa A. Abdelrheem, Noha A. Ahmed, Fahad M. Alshabrmi, Faris F. Aba Alkhayl, Maha A. Alwaili, Naif G. Altoom, Al Mokhtar Lamsabhi

{"title":"Mechanism-Based Allosteric Inhibition of PTP1B by Prenylated Flavonoids from Glycyrrhiza echinata: In Vitro Experiments and in Silico Validation","authors":"Emadeldin M. Kamel, Doaa A. Abdelrheem, Noha A. Ahmed, Fahad M. Alshabrmi, Faris F. Aba Alkhayl, Maha A. Alwaili, Naif G. Altoom, Al Mokhtar Lamsabhi","doi":"10.1007/s10930-025-10272-x","DOIUrl":"10.1007/s10930-025-10272-x","url":null,"abstract":"<div><p>A phytochemical investigation of <i>Glycyrrhiza echinata</i> led to the isolation and structural characterization of twelve phenolic compounds. An in silico target fishing analysis identified protein tyrosine phosphatase 1B (PTP1B) as a potential biological target for these phytochemicals, prompting an in vitro evaluation of their PTP1B inhibitory activities. Gancaonin Q and licoflavone C exhibited notably low IC₅₀ values (1.61 ± 0.32 µM and 1.39 ± 0.33 µM, respectively), outperforming the reference inhibitor ursolic acid (IC₅₀ = 7.17 ± 0.69 µM), while norartocarpetin showed moderate activity (IC₅₀ = 42.41 ± 2.12 µM). Enzyme kinetic studies revealed that gancaonin Q and licoflavone C act as noncompetitive inhibitors of PTP1B. Subsequent in silico analyses supported these findings and provided mechanistic insights. Molecular docking confirmed robust binding interactions for gancaonin Q and licoflavone C at the PTP1B allosteric site. Free energy landscape (FEL) calculations indicated that both compounds stabilized the enzyme within low-energy conformations, and MM/PBSA estimations corroborated their favorable binding free energies. Molecular dynamics simulations further demonstrated the stability of the ligand-enzyme complexes, characterized by reduced structural fluctuations in comparison with the free enzyme and norartocarpetin-bound states. Finally, ADMET assessments indicated promising pharmacokinetic and toxicity profiles, with some scope for structural refinement. Overall, these results highlight gancaonin Q and licoflavone C as promising lead compounds for the development of PTP1B inhibitors with therapeutic potential.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 5","pages":"526 - 549"},"PeriodicalIF":1.4,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144644497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Delineation of Recombinant Cestode Phosphoenolpyruvate Carboxykinase Activity Co-expressed with Molecular Chaperones 与分子伴侣共表达的重组Cestode磷酸烯醇丙酮酸羧激酶活性的描述。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-07-11 DOI: 10.1007/s10930-025-10279-4

Joplin Nongkhlaw, Superior Syngkli, Miranda Moirangthem, Bidyadhar Das

{"title":"Delineation of Recombinant Cestode Phosphoenolpyruvate Carboxykinase Activity Co-expressed with Molecular Chaperones","authors":"Joplin Nongkhlaw, Superior Syngkli, Miranda Moirangthem, Bidyadhar Das","doi":"10.1007/s10930-025-10279-4","DOIUrl":"10.1007/s10930-025-10279-4","url":null,"abstract":"<div><p>Phosphoenolpyruvate carboxykinase (PEPCK; EC 4.1.1.32) of cestodes is considered a possible anthelmintic target because of its differential role in their hosts. In an earlier study, the recombinant PEPCK from <i>Raillietina echinobothrida</i> (rePEPCK) was overexpressed as inclusion bodies and was solubilized following renaturation with chemical additives, specifically L-arginine. Molecular chaperones are alternatives to chemical additives and detergents because they preserve the stability and conformation of the proteins. Hence, in this study, the recombinant rePEPCK was subcloned into the pE-SUMO vector and co-expressed along with the molecular chaperones (e.g. pG-KJE8, pG-Tf2) in <i>Escherichia coli</i> BL21 (DE3) cells. The protein was purified using affinity chromatography and subsequently characterized. The overexpressed rePEPCK was found to be a monomer of ~ 75 kDa. The optimum activity of the enzyme was observed in 50 mM Tris–HCl buffer at pH 7.0. In comparison, Mn<sup>2+</sup> at 4.0 mM and GDP at 0.6 mM were observed to be the ideal cofactor and nucleotide, respectively. The <i>V</i><sub>max</sub> of the purified rePEPCK was found to be ~ 0.279 U/mg protein and <i>K</i><sub>m</sub> value of ~ 35.87 μM for its substrate. The turnover number (<i>k</i><sub>cat</sub>) of rePEPCK was found to be 4.7 s<sup>−1</sup> with catalytic efficiency (<i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub>) 1.31 × 10<sup>5</sup> M<sup>−1</sup> s<sup>−1</sup>. The chaperones interacted with the key amino acids of PEPCK. This investigation explored the role of the chaperones in producing biologically active rePEPCK for its characterisation and may improve the understanding of the biochemical and biophysical properties of the enzyme as an anthelmintic target.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"425 - 436"},"PeriodicalIF":1.4,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144610834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fluorescence Spectroscopy Reveals Resveratrol Binding to a Hydrophobic Pocket in Leishmania amazonensis Sir2-Related Protein 1 (LaSir2RP1) 荧光光谱揭示白藜芦醇与亚马逊利什曼原虫sir2相关蛋白1 （LaSir2RP1）疏水口袋的结合。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-22 DOI: 10.1007/s10930-025-10275-8

Melissa R. Fessel, Ana P. R. Povinelli, Veronica S. Fontes, Maria Isabel N. Cano, Carlos H. I. Ramos

{"title":"Fluorescence Spectroscopy Reveals Resveratrol Binding to a Hydrophobic Pocket in Leishmania amazonensis Sir2-Related Protein 1 (LaSir2RP1)","authors":"Melissa R. Fessel, Ana P. R. Povinelli, Veronica S. Fontes, Maria Isabel N. Cano, Carlos H. I. Ramos","doi":"10.1007/s10930-025-10275-8","DOIUrl":"10.1007/s10930-025-10275-8","url":null,"abstract":"<div><p>Resveratrol, a natural phytoalexin synthesized by certain plants in response to injury, exhibits antimicrobial properties and various medically significant effects, including anticancer and antiaging activities. Although its exact mechanism of action is still under investigation, it is believed to involve its interaction with a group of protein deacetylases known as sirtuins. Sirtuins are crucial in regulating metabolism, stress responses, and processes such as lifespan extension through caloric restriction. In this study, we report the inhibitory effect of resveratrol on the growth of <i>Leishmania amazonensis</i> promastigotes, highlighting its potential as a microbicide. Through fluorescence spectroscopy assays and in silico analysis, we identified and characterized the interaction between resveratrol and Sir2-related protein 1 from <i>L. amazonensis</i> (rLaSir2RP1). Our results demonstrate a direct interaction between resveratrol and rLaSir2RP1, characterized by a binding constant of 10<sup>5</sup> M<sup>−1</sup>. This interaction involves a single binding site located near a hydrophobic pocket, which includes its solely tryptophan residue.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"387 - 398"},"PeriodicalIF":1.4,"publicationDate":"2025-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144370026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-Yield Biosynthesis Process for Producing Insulin Lispro 胰岛素Lispro高产生物合成工艺研究

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-22 DOI: 10.1007/s10930-025-10278-5

Shuchen Pei, Shusheng Liu, Jihong Luo, Junlin Chen, Hua Luo, Wen Wu, Xinan Chen

{"title":"High-Yield Biosynthesis Process for Producing Insulin Lispro","authors":"Shuchen Pei, Shusheng Liu, Jihong Luo, Junlin Chen, Hua Luo, Wen Wu, Xinan Chen","doi":"10.1007/s10930-025-10278-5","DOIUrl":"10.1007/s10930-025-10278-5","url":null,"abstract":"<div><p>Insulin lispro, a fast-acting insulin analogue, is widely used for the treatment of diabetes due to its rapid onset and high flexibility. This study established a three-step chromatography purification process, L fermentation broth can obtain 475 mg/L insulin lispro. Through a multi-step process involving fermentation, recovery of inclusion bodies, renaturation, enzymatic digestion, and chromatographic purification, a high-purity insulin lispro product was obtained. The final product achieved a purity of 99.7%. The high molecular weight protein was ≤ 0.1%, zinc content was 0.41%, host cell protein contamination was 2.31 ng/mg, and residual DNA was 1.57 ng/mg. Analytical results, including retention time, molecular weight, and peptide mapping, were consistent with theoretical expectations and matched the control. This purification process provides a streamlined and cost-effective method for large-scale production of insulin lispro, supporting further development of recombinant insulin analogs and offering high-quality active pharmaceutical ingredients for pharmaceutical formulations.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"363 - 376"},"PeriodicalIF":1.4,"publicationDate":"2025-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144370027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correction: From Crude Extracts to Purity: A Comparative Study of Butyrylcholinesterase Purification 更正：从粗提取物到纯度：丁基胆碱酯酶纯化的比较研究。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-14 DOI: 10.1007/s10930-025-10274-9

Gamze Sonmez, Bahattin Enes Karatas, Ebru Bodur

引用次数: 0

Purification and Characterization of a New Phytolectin Extracted from Eucalyptus vicina Fruits with Immunomodulatory Effect 一种具有免疫调节作用的桉叶植物素的纯化及特性研究。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-11 DOI: 10.1007/s10930-025-10277-6

Sana Boufeker, Ahlem Bahi, Imene Torche, Yacer Boudersa, Ali Shaikh-Ibrahim, Monna Taki Ellh Bousaoula, Sondos Hejazi

{"title":"Purification and Characterization of a New Phytolectin Extracted from Eucalyptus vicina Fruits with Immunomodulatory Effect","authors":"Sana Boufeker, Ahlem Bahi, Imene Torche, Yacer Boudersa, Ali Shaikh-Ibrahim, Monna Taki Ellh Bousaoula, Sondos Hejazi","doi":"10.1007/s10930-025-10277-6","DOIUrl":"10.1007/s10930-025-10277-6","url":null,"abstract":"<div><p>Plant-derived lectins are getting growing interest for their potential biomedical applications, however, there are few studies on lectins from <i>Eucalyptus vicina</i> fruits. The aim of the current study was to isolate and characterize a new phytolectin from <i>Eucalyptus vicina</i> fruits (EVL) and evaluate its immunomodulatory activity. EVL was purified through ammonium sulfate precipitation (70%) followed by ion-exchange chromatography. The purified protein appeared to have a molecular weight of around 55 kDa, determined using sodium dodecyl sulfate–polyacrylamide gel electrophoresis SDS-PAGE. EVL had strong hemagglutination activity against rabbit erythrocytes and all human blood types and displayed a high affinity against fetuin. The lectin exhibited optimal activity at alkaline pH (8–10) and was remarkably thermostable for 1 h at 70 degrees. Metal ions did not alter activity to a significant degree. The immunomodulatory effects were assessed in vivo using a carbon clearance assay, and EVL injection resulted in a dose-dependent increase in phagocytic activity. Overall, our data suggests that EVL could be a valuable candidate for research and future immunomodulatory therapies.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"351 - 362"},"PeriodicalIF":1.4,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144268321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification of Potential RNAi Targets for Cotton Mealybug (Phenacoccus solenopsis Tinsley) Management 棉花粉蚧（Phenacoccus solenopsis Tinsley）管理潜在RNAi靶点的鉴定

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-06 DOI: 10.1007/s10930-025-10269-6

Sanchita Singh, Somnath Rahangdale, Shivali Pandita, Manisha Singh, Gauri Saxena, Gourav Jain, Praveen C. Verma

{"title":"Identification of Potential RNAi Targets for Cotton Mealybug (Phenacoccus solenopsis Tinsley) Management","authors":"Sanchita Singh, Somnath Rahangdale, Shivali Pandita, Manisha Singh, Gauri Saxena, Gourav Jain, Praveen C. Verma","doi":"10.1007/s10930-025-10269-6","DOIUrl":"10.1007/s10930-025-10269-6","url":null,"abstract":"<div><p><i>Phenacoccus solenopsis</i> Tinsley (Hemiptera: Pseudococcidae), commonly known as the cotton mealybug, is a highly invasive pest worldwide, particularly in tropical and subtropical regions. Despite posing a significant economic threat to various agricultural crops, a promising, environmentally friendly control strategy against this pest is lacking. Furthermore, the molecular aspects of this insect pest remain understudied. This pioneering study provides the first proteome data for four different developmental stages of the cotton mealybug. A comparative analysis of differential protein expression (DEPs) among six groups revealed the highest number of DEPs (550 up-regulated and 1118 down-regulated) when comparing the egg and first nymphal stages to the second nymphal instar (fold change ≥ 2, <i>P</i> < 0.05). From the generated proteomics data, potential target genes were identified for cotton mealybug management. These genes were further evaluated for RNAi-based pest control and optimization of the dsRNA delivery system in cotton mealybug. Notably, RNAi-based pest management analysis revealed that dsRNA of the Ferritin-like precursor (Psfer) gene (TRINITY_DN17055_c1_g1_i1) induced significant mortality (~ 69%), followed by dsRNA of the probable cytochrome P450 6a14-like (Psp450 6a14) gene (TRINITY_DN47081_c0_g1) and odorant-binding protein 2 precursor (Psobp) gene (TRINITY_DN11547_c0_g1). This investigation proposes a potential alternative, eco-friendly strategy for managing cotton mealybug populations and related pests. Furthermore, this study provides valuable insights into the proteome of the cotton mealybug and Hemiptera, offering avenues for proteome-based identification of RNAi targets for pest management and crop improvement.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"464 - 481"},"PeriodicalIF":1.4,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144236382","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Expression and Purification of Human Stem Cell Factor and Its Effect on the Growth of A549 Cells 人干细胞因子的表达纯化及其对A549细胞生长的影响。

IF 1.4 4区生物学

The Protein Journal Pub Date : 2025-06-02 DOI: 10.1007/s10930-025-10273-w

Zhenling Ma, Xulu Han, Yiyang Zhu, Panpan Fu, Zhenzhen Li, Yongqin Zhao, Junzheng Yang, Wei Liu

{"title":"Expression and Purification of Human Stem Cell Factor and Its Effect on the Growth of A549 Cells","authors":"Zhenling Ma, Xulu Han, Yiyang Zhu, Panpan Fu, Zhenzhen Li, Yongqin Zhao, Junzheng Yang, Wei Liu","doi":"10.1007/s10930-025-10273-w","DOIUrl":"10.1007/s10930-025-10273-w","url":null,"abstract":"<div><p>Stem cell factor (SCF) is a type of hematopoietic cytokine that functions in early hematopoietic stem cells. SCF, together with its cognate receptor c-kit, plays crucial roles in cell survival, proliferation, and differentiation. A dysregulated SCF/c-kit system is reportedly, associated strongly with various kinds of cancer in humans. However, the function and mechanism of SCF in non-small cell lung cancer (NSCLC) remains to be elucidated to date. In this context, the present study attempted to purify two different sizes of human SCF proteins and then investigate the effect of these exogenous SCFs in A549 cells. First, the <i>homo sapiens</i> Kit ligand (KITG) transcript variants a and b were amplified, and recombinant pET-30a vectors were constructed. Soluble His-SCF and His-SCF-2 were then expressed and purified using 0.1 mM isopropyl-β-D-1-tiogalactopiranoside (IPTG) at 25 °C and 0.3 mM IPTG at 25 °C, respectively. MTT assay was conducted next, which revealed that the purified proteins significantly promoted A549 cell activity and increased the phosphorylation of P65. Further, NF-κB pathway blockade was achieved through treatment with BAY11-7082, and it was observed to decrease the A549 cell activity promoted by the purified proteins. These findings provided insights into the biological role of SCF in the development of NSCLC.</p></div>","PeriodicalId":793,"journal":{"name":"The Protein Journal","volume":"44 4","pages":"377 - 386"},"PeriodicalIF":1.4,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144210615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0