Sheng wu gong cheng xue bao = Chinese journal of biotechnology最新文献

[Signature motif identification and enzymatic characterization of a protein tyrosine phosphatase in Metarhizium anisopliae]. [金龟子绿僵菌蛋白酪氨酸磷酸酶的特征基序鉴定和酶学表征]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250124

Ze Tan, Pei Zhu, Zhenlun Li, Shuiying Yang

{"title":"[Signature motif identification and enzymatic characterization of a protein tyrosine phosphatase in Metarhizium anisopliae].","authors":"Ze Tan, Pei Zhu, Zhenlun Li, Shuiying Yang","doi":"10.13345/j.cjb.250124","DOIUrl":"https://doi.org/10.13345/j.cjb.250124","url":null,"abstract":"Protein tyrosine phosphatases (PTPs, EC 3.1.3.48) are key regulators of cellular processes, with the catalytic activity attributed to the conserved motif (H/V)CX5R(S/T), where cysteine and arginine residues are critical. Previous studies revealed that alternative splicing of extracellular phosphatase mRNA precursors in Metarhizium anisopliae generated two distinct transcripts, with the longer sequence containing a novel HCPTPMLS motif resembling PTP signatures but lacking the arginine residue. To identify the novel signature motif and characterize its enzymatic properties, we heterologously expressed and purified both proteins in Pichia pastoris and comprehensively characterized their enzymatic properties. The protein containing the HCPTPMLS motif (designated as L-protein) exhibited the highest activity at pH 5.5 and a strong preference for pTyr substrates. Its phosphatase activity was inhibited by Ag+, Zn2+, Cu2+, molybdate, and tungstate, but enhanced by Ca2+ and EDTA. AcP101 (lacking HCPTPMLS) showed the maximal activity at pH 6.5 and a strong preference toward pNPP (P < 0.05), with the activity inhibited by NaF and tartrate, but enhanced by Mg2+ and Mn2+. Functional analysis confirmed that the L-protein retained the PTP activity despite the absence of arginine in its signature motif, while AcP101 functioned as an acid phosphatase. This study provides the first functional validation of an arginine-deficient PTP motif, expanding the definition of PTP signature motifs and offering new insights for phosphatase classification.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3579-3588"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138804","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

[Advances in yeast biosynthesis of triterpenoids for cosmetic applications]. 酵母生物合成化妆品用三萜的研究进展

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250116

Yilin Li, Shuai Wang, Ying Wang, Chun Li

{"title":"[Advances in yeast biosynthesis of triterpenoids for cosmetic applications].","authors":"Yilin Li, Shuai Wang, Ying Wang, Chun Li","doi":"10.13345/j.cjb.250116","DOIUrl":"https://doi.org/10.13345/j.cjb.250116","url":null,"abstract":"Triterpenoids in cosmetic raw materials have attracted much attention due to their various skin-care effects such as anti-inflammatory, antioxidant, and moisturizing properties, showing broad application prospects. However, the conventional methods such as chemical synthesis and plant extraction for obtaining triterpenoid have problems like poor sustainability, which limit their application in large-scale production. In recent years, with the development of synthetic biology and metabolic engineering, yeast synthesis of compounds has provided a green and sustainable alternative for the production of triterpenoids. This article reviews the research progress in the synthesis of triterpenoids and their derivatives in the cosmetic field and elaborates on the two main synthesis pathways (mevalonate and methylerythritol phosphate pathways) and their advantages and limitations in different microbial hosts. In addition, this article introduces the current status of the synthesis of triterpenoids and their derivatives in yeast, discusses the current strategies for increasing the yield, and looks ahead to the future development directions, aiming to promote the applications of triterpenoids in the cosmetic field.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3405-3425"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138610","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

[Whole-cell catalytic synthesis of β-hydroxy-β-methylbutyric acid by constructing recombinant Escherichia coli]. [重组大肠杆菌全细胞催化合成β-羟基-β-甲基丁酸]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250243

Jiawei Ye, Hong Xu, Yaxin Liao, Zhiming Rao, Meijuan Xu

{"title":"[Whole-cell catalytic synthesis of β-hydroxy-β-methylbutyric acid by constructing recombinant Escherichia coli].","authors":"Jiawei Ye, Hong Xu, Yaxin Liao, Zhiming Rao, Meijuan Xu","doi":"10.13345/j.cjb.250243","DOIUrl":"https://doi.org/10.13345/j.cjb.250243","url":null,"abstract":"β-hydroxy-β-methylbutyric acid (HMB) is widely applied in sports nutrition, disease prevention and other fields. However, chemical synthesis methods, limited by toxic reagents and violent reactions, can hardly meet the demands of green production. The biosynthesis method mainly utilizes enzymatic catalysis or metabolic engineering techniques for synthesis, and has the advantages of high efficiency, low cost, and sustainability. Therefore, the production of HMB by the biosynthesis method has a good application prospect. In this research, a biosynthesis-based production strategy for HMB was developed. By using L-leucine as the substrate and constructing a dual-enzyme co-expression system, we established an efficient catalytic process. At first, the enzymatic properties of L-amino acid deaminase (PvL-AAD) from Proteus vulgaris and 4-hydroxyphenylpyruvate dioxygenase (Rn4-HPPD) from Rattus norvegicus were characterized. Rn4-HPPD had low relative activity and required an acidic environment for catalysis. Based on the surface charge modification strategy of the enzyme protein, site-directed mutagenesis and combinatorial mutagenesis were conducted on 10 sites of Rn4-HPPD. A double mutant Rn4-HPPDH18R/N302R was thus obtained, with the enzyme activities being 2.00 times and 2.39 times that of the wild type at pH 5.5 and pH 6.5, respectively. Subsequently, the expression of the two enzymes in Escherichia coli was optimized. After the optimal expression ratio of the two enzymes was determined as 1:3 and under the conditions of OD600 of 70, pH 6.0, 35 ℃, Fe2+ concentration of 1.5 mmol/L, and feeding of the substrate in batches in a 5 L fermenter, the maximum yield of HMB reached 8.60 g/L. This study not only enhances the optimal pH and activity of Rn4-HPPD but also provides new approaches for the efficient microbial synthesis of HMB.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3487-3503"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138845","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

[Synthetic promoters: theory, design, and prospects]. [合成助剂：理论、设计与展望]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250044

Peng Peng, Minghai Chen, Qin Li, Xian'en Zhang

{"title":"[Synthetic promoters: theory, design, and prospects].","authors":"Peng Peng, Minghai Chen, Qin Li, Xian'en Zhang","doi":"10.13345/j.cjb.250044","DOIUrl":"https://doi.org/10.13345/j.cjb.250044","url":null,"abstract":"Synthetic promoters are novel promoters artificially designed and do not exist in nature. They can initiate the expression of target genes with specific regulatory modes, offering advantages such as high expression efficiency, precise regulation, and modularity. These features endow synthetic promoters with significant application potential in fields such as industrial production, environmental monitoring, and disease diagnosis and treatment. This paper reviews the basic structures, functions, and classification of promoters, discusses various regulatory elements that influence promoter functions, including enhancers, signal response elements, and transcription factors. Additionally, the conventional and deep learning-based strategies for designing synthetic promoters are summarized. Finally, the theoretical significance of synthetic promoters is emphasized, which is followed by an overview of their current applications, along with a rational discussion on the challenges and future development directions of synthetic promoters. Given the critical role of promoters in gene regulation, this article provides a review and outlook on the research progress of synthetic promoters, which holds reference value for the design of cellular gene circuits.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3351-3374"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138900","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

[Establishment and optimization of a genetic manipulation system for Staphylococcus pasteuri]. 巴氏葡萄球菌基因操作系统的建立与优化

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250167

Tinghao Zhang, Ziqi Wang, Yuxin Song, Jinjin Wang, Feng Guo, Yongjun Zhang, Fuping Lu, Ming Li

{"title":"[Establishment and optimization of a genetic manipulation system for Staphylococcus pasteuri].","authors":"Tinghao Zhang, Ziqi Wang, Yuxin Song, Jinjin Wang, Feng Guo, Yongjun Zhang, Fuping Lu, Ming Li","doi":"10.13345/j.cjb.250167","DOIUrl":"https://doi.org/10.13345/j.cjb.250167","url":null,"abstract":"One of the technical bottlenecks limiting the high yield of 1,4-butanediamine is the insufficient tolerance of strains to 1,4-butanediamine. Enhancing the tolerance of strains to 1,4-butanediamine is therefore a primary challenge that needs to be addressed for the construction of strains with high yields of 1,4-butanediamine. Staphylococcus pasteuri 326180 exhibits exceptional tolerance to high-concentration 1,4-butanediamine, serving as both an ideal model for studying the mechanism underlying the 1,4-butanediamine tolerance and a novel host for constructing strains capable of efficiently producing 1,4-butanediamine. However, for both the research on the tolerance mechanism and the modification of chassis strains, gene editing of S. pasteuri needs to be carried out at the molecular level. The research objective of this paper is to establish a genetic manipulation system for S. pasteuri, laying foundation for subsequent studies on tolerance mechanism and the modification of chassis strains. This study systematically optimized the electroporation conditions, including key parameters such as the growth phase of cells, electric field strength, electroporation buffer, and recovery medium, successfully establishing an electroporation method for S. pasteuri. Additionally, we constructed the gene editing plasmid pCpfOA by replacing the resistance expression cassette, optimized the selection markers for gene editing, and finally established a CRISPR/Cpf1-based gene editing technology for S. pasteuri, achieving an editing efficiency of 90%. The genetic manipulation system of S. pasteuri established in this study provides technical support for research into the tolerance mechanism of this bacterium and the genetic modification of chassis strains.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3604-3616"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138786","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

[Mechanisms and applications of microbial synthesis of metal nanoparticles]. [微生物合成金属纳米颗粒的机理及应用]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250224

Xinruo Wang, Chaoning Hu, Yangyang Wang, Aoqi Song, Rui Tang, Feng Li, Hao Song

{"title":"[Mechanisms and applications of microbial synthesis of metal nanoparticles].","authors":"Xinruo Wang, Chaoning Hu, Yangyang Wang, Aoqi Song, Rui Tang, Feng Li, Hao Song","doi":"10.13345/j.cjb.250224","DOIUrl":"https://doi.org/10.13345/j.cjb.250224","url":null,"abstract":"The rapid growth of electronic waste has led to the accumulation of large amounts of valuable metal elements in the environment, causing serious environmental pollution and resource wastage. Compared with pyrometallurgical and hydrometallurgical processes which often result in severe environmental pollution and carbon footprints, microbial synthesis of metal nanoparticles has emerged as a green and environmentally friendly metallurgical technology for recovering valuable metals from electronic waste. This paper first reviews the mechanisms of metal nanoparticle synthesis within different structural compartments of microbial cells. It then introduces the applications of microbially synthesized metal nanoparticles in fields such as environmental remediation, energy production, biocatalysis, and biomedicine. Finally, it discusses the development prospects of microbial synthesis of metal nanoparticles, including exploration of diverse microbial resources and synthesis pathways, yield enhancement, integration of new technologies, and industrialization, aiming to promote further research and application of microbial synthesis of metal nanoparticles.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3387-3404"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138805","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

[High-throughput screening technologies in the engineering of actinomycete strains]. 放线菌菌株工程中的高通量筛选技术

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250267

Xueyan Liu, Meng Wang, Jifeng Liu, Yue Zhang

{"title":"[High-throughput screening technologies in the engineering of actinomycete strains].","authors":"Xueyan Liu, Meng Wang, Jifeng Liu, Yue Zhang","doi":"10.13345/j.cjb.250267","DOIUrl":"https://doi.org/10.13345/j.cjb.250267","url":null,"abstract":"Actinomycetes are important producers of high-value natural products, and the engineering of actinomycetes to enhance the biosynthesis of target natural products has long been a hot research topic in the scientific community. However, non-rational engineering methods suffer from low beneficial mutation rates, which limit the efficiency of mutant screening. The integration of high-throughput screening (HTS) technologies can effectively enhance the screening efficiency of elite mutants and significantly shorten the cycle of actinomycete strain engineering. This review comprehensively discusses various HTS technologies suitable for the engineering of actinomycete strains and compares them in terms of application scenarios, advantages, and disadvantages. HTS technologies include microplate-based screening, antimicrobial activity screening, antibiotic resistance screening, fluorescence-activated cell sorting (FACS), and fluorescence-activated droplet sorting (FADS). Additionally, this review summarizes the applications of these technologies in assisting actinomycete strain engineering and enhancing the yields of target compounds. The development and application of HTS technologies have not only facilitated the exploration of natural product resources in actinomycetes but also provided strong support for the rapid and efficient construction of high-performance engineered actinomycete strains.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3375-3386"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138878","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

[Discovery and protein engineering of penicillin G acylase for biosynthesis of cefradine]. [生物合成头孢拉定用青霉素G酰化酶的发现及蛋白工程]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250265

Lingyi Liu, Xiangying Li, Congcong Li, Lijuan Ma, Bo Yuan, Zhoutong Sun

引用次数: 0

[Protein engineering for the modification of a L-amino acid deaminase for efficient synthesis of phenylpyruvic acid]. [为高效合成苯丙酮酸而对l -氨基酸脱氨酶进行修饰的蛋白质工程]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250075

Xuanping Shi, Yue Wang, Zhina Qiao, Jiajia You, Zhiming Rao

{"title":"[Protein engineering for the modification of a L-amino acid deaminase for efficient synthesis of phenylpyruvic acid].","authors":"Xuanping Shi, Yue Wang, Zhina Qiao, Jiajia You, Zhiming Rao","doi":"10.13345/j.cjb.250075","DOIUrl":"https://doi.org/10.13345/j.cjb.250075","url":null,"abstract":"Phenylpyruvic acid (PPA) is used as a food and feed additive and has a wide range of applications in the pharmaceutical, chemical and other fields. At present, PPA is mainly produced by chemical synthesis. With the green transformation of the manufacturing industry, biotransformation will be a good alternative for PPA production. The L-amino acid deaminase (PmiLAAD) from Proteus mirabilis has been widely studied for the production of PPA. However, the low yield limits its industrial production. To further enhance the production of PPA and better meet industrial demands, a more efficient synthesis method for PPA was established. In this study, PmiLAAD was heterologously expressed in Escherichia coli. Subsequently, a colorimetric reaction method was established to screen the strains with high PPA production. The semi-rational design of PmiLAAD was carried out, and the obtained triple-site mutant V18 (V437I/S93C/E417A) showed a 35% increase in catalytic activity compared with the wild type. Meanwhile, the effect of N-terminal truncation on the catalytic activity of the V18 mutant was investigated. After the optimization of the whole-cell conditions for the obtained mutant V18-N7, fed-batch conversion was carried out in a 5-L fermenter, and 44.13 g/L of PPA was synthesized with a conversion rate of 88%, which showed certain potential for industrial application. This study lays foundation for the industrial production of phenylpyruvic acid and also offers insights into the biosynthesis of other chemicals.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3521-3536"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138879","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

[A multi-enzyme cascade process for the preparation of L-phosphinothricin]. [一种多酶级联工艺制备l -膦黄芩苷]。

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-09-25 DOI: 10.13345/j.cjb.250199

Manman Wang, Yu Yang, Xianbing Song, Xiaolian Li, Binchun Li, Ziqiang Wang

{"title":"[A multi-enzyme cascade process for the preparation of L-phosphinothricin].","authors":"Manman Wang, Yu Yang, Xianbing Song, Xiaolian Li, Binchun Li, Ziqiang Wang","doi":"10.13345/j.cjb.250199","DOIUrl":"https://doi.org/10.13345/j.cjb.250199","url":null,"abstract":"L-phosphinothricin (L-PPT) is an efficient broad-spectrum herbicide. To realize the multi-enzyme catalytic preparation of L-PPT, we constructed an engineered strain Escherichia coli YM-1 for efficient expression of D-amino acid transaminase, which could catalyze the generation of the intermediate 2-oxo-4-[(hydroxymethylphosphonyl)] butyric acid (PPO) from D-phosphinothricin (D-PPT). In addition, E. coli pLS was constructed to co-express glutamate dehydrogenase and glucose dehydrogenase, which not only catalyzed the generation of L-PPT from PPO but also regenerated the coenzyme nicotinamide adenine dinucleotide phosphate (NADPH). A fed-batch fermentation process was then established for E. coli YM-1 and pLS, and the apparent activities of D-amino acid transaminase and glutamate dehydrogenase were increased by 22.68% and 100.82%, respectively, compared with those in shake flasks. The process parameters were optimized for the catalytic preparation of L-PPT by whole-cell cascade of E. coli YM-1 and pLS with D, L-PPT as the substrate. After reaction for 8 h, 91.36% conversion of D-PPT was achieved, and the enantiomeric excess of L-PPT reached 90.22%. The findings underpin the industrial production of L-PPT.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 9","pages":"3589-3603"},"PeriodicalIF":0.0,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145138607","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