Biotechnology and Bioengineering最新文献

{"title":"Microtiter Plate Cultivation Systems Enable Chemically Diverse Metabolic Footprints During Bacterial Natural Product Discovery","authors":"Anton Lindig, Georg Hubmann, Stephan Lütz","doi":"10.1002/bit.29002","DOIUrl":"https://doi.org/10.1002/bit.29002","url":null,"abstract":"Rediscovery of known structures is a frequent problem in screening for bioactive bacterial natural products (NPs). Highly parallelized microtiter plate cultivation systems (MPCS) can improve the chance to discover novel NPs by testing a multitude of cultivation conditions simultaneously. An in‐depth analysis and comparison of cultivation systems for NP discovery, however, has not been carried out so far. We compared the growth and metabolic footprint of four distinct bacterial species in three MPCS, shake flasks, and stirred tank bioreactors (STR). While the big majority of the cultivation systems provided good growth, we found a considerable divergence in secondary metabolite (SM) formation. The SM space was approximated by the appearance of unique mass features (MFs) in the supernatant extracts throughout the cultivation period. Molecular network analysis was applied to visualize the changes from detected MFs at the molecular level. The cultivation systems had a minor impact on the unicellular growing <jats:italic>Bacillus amyloliquefaciens</jats:italic>. This impact was more pronounced for the tested filamentous bacteria, resulting in a diversified metabolic footprint. The maximal overlap of 31% of produced MFs indicates a lack of comparability between the cultivation systems, resulting in different entries of growth phases and the formation of associated SMs. The detected SMs and its derivatives exhibited structural modification depending on the cultivation system. A comparison of <jats:italic>Streptomyces griseochromogenes</jats:italic> NP profile revealed that MPCS yielded less divergent SM formation than shake flasks. Our comprehensive assessment is the first to demonstrate the impact of cultivation systems on the bacterial metabolic footprint, confirming that MPCS provide a robust platform for the parallelization of bacterial cultivations for the discovery of bacterial NPs and accessing the chemical NP space more broadly.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"78 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Mathematical Model for Determining Probabilistic Design Space in Mesenchymal Stem Cell Passage Culture","authors":"Keita Hirono, Yusuke Hayashi, Yuuki Ogawa, Masahiro Kino-oka, Hirokazu Sugiyama","doi":"10.1002/bit.29001","DOIUrl":"https://doi.org/10.1002/bit.29001","url":null,"abstract":"With their many therapeutic functions, mesenchymal stem cells (MSCs) are promising sources for regenerative medicine. However, in the manufacture of MSCs, without a method for exploring the effects of long-term passage on cell proliferation potentials, the design of passage culture processes is challenging. Here, for the process design of the MSC passage culture, we propose a model for predicting the growth rate as a function of the cumulative population doubling level (cPDL) for each passage. Three steps were implemented: (1) passage culture experiments to correlate apparent growth rate with cPDL were conducted, (2) a model for predicting the growth rate as a function of cPDL was developed, and (3) a model to design the passage culture of MSCs from bone marrow (BM-MSCs) and umbilical cord (UC-MSCs) with stochastic simulation was applied. Two design variables (passage number and harvesting time) were investigated to define feasible operation regions as probabilistic design spaces to meet three quality indicators (senescence level, confluency level, and total number of cells) with given probabilities. Consequently, 10 and 62 conditions out of 165 were identified as feasible for BM- and UC-MSCs, respectively, which would contribute to the industrial MSC passage culture process design.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"52 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143872520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic Remodeling of Mechano‐Sensing Complexes in Suspended Fibroblast Cell‐Sheets Under External Mechanical Stimulus","authors":"Madoka Suzuki, Keiko Kawauchi, Hiroaki Machiyama, Hiroaki Hirata, Shin'ichi Ishiwata, Hideaki Fujita","doi":"10.1002/bit.28996","DOIUrl":"https://doi.org/10.1002/bit.28996","url":null,"abstract":"Freestanding cell‐sheets are valuable bio‐materials for use in regenerative medicine and tissue engineering. Because cell‐sheets experience various mechanical stimulations during handling, it is important to understand the responses of cells to these stimulations. Here, we demonstrate changes in the localization of various proteins during the stretching of fibroblast cell‐sheets. These proteins are known to be involved in mechano‐sensing. Upon stretching, actin filaments appear parallel to the stretching direction. At cell‐cell junctions, β‐catenin forms clusters that co‐localize with accumulated vinculin and zyxin as well as the actin filaments. The p130 Crk‐associated substrate, known to be present in focal adhesions, is also recruited to these clusters and phosphorylated. Our results suggest that mechano‐sensing machinery is formed at cell‐cell junctions when the cell‐sheets are stretched.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"7 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143866982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combinatorial Metabolic Engineering for Enhanced Gibberellic Acid Biosynthesis in Fusarium fujikuroi.","authors":"Xia Ke,Yao Chen,Rui Jia,Hao-Nan Wang,Zhi-Qiang Liu,Yu-Guo Zheng","doi":"10.1002/bit.29005","DOIUrl":"https://doi.org/10.1002/bit.29005","url":null,"abstract":"Gibberellic acid (GA3), a quintessential diterpenoid phytohormone, is indispensable in agronomic practices, horticulture, and the wine industry. This study implemented a combinatorial metabolic engineering strategy within Fusarium fujikuroi (F. fujikuroi) by integrating the potentiation of global regulatory factors (GRFs), and amplification of biosynthetic precursors, alongside dynamic modulation of cofactors with dissolved oxygen supply, to precisely enhance GA3 biosynthesis. Transcriptomic analyses revealed that positive GRFs (AreB, Hat1, and Ada3) enhanced carbon and nitrogen metabolism, increased biomass accumulation, and upregulated transcription levels of the GA3 biosynthetic gene cluster. The use of endogenous nitrogen-responsive promoters ensured a balanced supply of cofactors and oxygen, thereby preventing the accumulation of terpenoid by-products. These combinatorial metabolic engineering strategies presented in this study make a significant step toward the enhancement of GA3 yield (3.22 g/L) via submerged fermentation of F. fujikuroi, offering novel insights to enable high-level biosynthesis of secondary metabolites in fungal chassis.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"24 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143872021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling Analysis of Oxygen Transfer Efficiency in Rest Cell Catalysis for Extra High-Titer Xylonic Acid Bioproduction","authors":"Xia Hua, Yong Xu","doi":"10.1002/bit.29004","DOIUrl":"https://doi.org/10.1002/bit.29004","url":null,"abstract":"The conflict arising from high-titer products and substantial oxygen requirements in aerobic bioconversion results in high-viscosity and oxygen transfer bottlenecks in dynamically changing biosystems. Currently, in the bioproduction of xylonic acid (XA), strategies to address the oxygen transfer bottleneck predominantly focus on macro-level modifications of the bioreactor. In this study, aiming at the high-viscosity biosystem, the optimal rotational speed equation was established at the fluid level by quantitatively investigating the variations and limitations of fluid rheological characteristics, gas holdup, cell respiration rate, and volume transfer coefficient of broth under different concentrations and rotational speeds. Based on the cell respiration rate under the optimal rotation speed, the theoretical production performance was calculated, and 679.3 g/L XA was achieved with the productivity of 14.2 g/L/h by batch feeding mode. Verified using actual production under the same conditions as a control, 649.3 g/L XA was finally accumulated with a productivity of 13.5 g/L/h, which was equivalent to 95.8% of the theoretical production. The intensification strategy for oxygen transfer provided insightful ideas to overcome the stubborn obstacles of obligate aerobic catalysis. Moreover, the study offered technical assistance and application potential for the production of high-titer XA from high-viscosity sugar-rich lignocellulosic hydrolysate.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"7 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143867042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimized Biosynthetic Pathway for Nonnatural Amino Acids: An Efficient Approach for L-2-Aminobutyric Acid Production","authors":"Jianmiao Xu, Yuan Tao, Qilan Shan, Yan Feng, Yihong Wang, Zhiqiang Liu, Yuguo Zheng","doi":"10.1002/bit.29003","DOIUrl":"https://doi.org/10.1002/bit.29003","url":null,"abstract":"L-2-Aminobutyric acid (L-2-ABA) is a nonnatural chiral α-amino acid which is widely used in various chiral pharmaceuticals and medical intermediates. Currently, the microbial metabolic engineering approach to enable <i>Escherichia coli</i> to produce L-2-ABA autonomously exists the problem of low synthesis efficiency, limiting its large-scale application. In this study, we successfully constructed a strain of <i>E. coli</i> that can produce L-2-ABA efficiently via multi-pathway transformation. Firstly, the growth defect of the start strain was restored by the help of screening transcriptional regulators. To maximize the accumulation of L-2-ABA, enhancements were made to the main synthetic pathways as well as cofactor systems and energy supply. Subsequently, transport proteins associated with osmotic stress tolerance were modified to improve adaptability of the strain during fermentation. Ultimately, the titer of L-2-ABA reached 42.14 g/L through the final strain ABAT38 in a 5-liter bioreactor, with a productivity of 0.40 g/L/h and a glucose conversion of 0.39 g/g, which exceeded the highest levels reported before. The strategies proposed in this study contribute to the production of L-2-ABA. At the same time, it has reference significance for the biosynthesis of related nonnatural amino acids with phosphoenolpyruvate as the intermediate metabolite.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"17 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143862090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrating Affinity Chromatography in the Platform Process for Adenovirus Purification","authors":"Yuxuan Wu, Eduardo Barbieri, William K. Smith, Arianna Minzoni, Ryan E. Kilgore, Wenning Chu, Michael A. Daniele, Stefano Menegatti","doi":"10.1002/bit.29006","DOIUrl":"https://doi.org/10.1002/bit.29006","url":null,"abstract":"Adenoviral vectors (AdVs) are gaining prominence in cancer therapy and vaccine development, posing the need for a modern AdV manufacturing platform. Current AdV purification by ion-exchange chromatography indeed struggles to achieve the product's yield and purity of processes that employ affinity technologies. Addressing these challenges, this study presents the first affinity-based process that delivers high product yield and clearance of host cell proteins and DNA (HCPs and hcDNA) in two chromatography steps. The affinity capture utilizes resins functionalized with peptide ligands that target AdV hexon proteins (AEFFIWNA and TNDGPDYSSPLTGSG), and provide high capacity (&gt; 5·10¹⁰ vp/mL of resin) and yield under mild elution conditions (~50% at pH 8.0). Peptide-functionalized adsorbents prepared using different matrices (polymethylmethacrylate vs. agarose) were initially tested to compare the purification performance. AEFFIWNA-SulfoLink resin was selected for its yield of cell-transducing AdVs (~50%) and removal of HCPs and hcDNA (144-fold and 56-fold). Similarly, TNDGPDYSSPLTGSG-Toyopearl resin afforded ~50% yield and &gt; 50-fold reduction of impurities. Additional gains in product purity were achieved by optimizing the washing step, which removed free hexon proteins and additional HCPs. All peptide-functionalized resins maintained their purification performance for 10 cycles upon regeneration at pH ~2.0. The purification process was assembled to include clarification, affinity capture in bind-and-elute mode using AEFFIWNA-SulfoLink resin, and polishing in flow-through mode using mixed-mode resins. The optimized process provided a yield ~50% of cell-infecting units (IFU) and a product titer ~10⁷ IFU/mL, along with residual HCP and hcDNA levels (8.76 ng/mL and 44 ng per dose, respectively) that meet clinical requirements.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"33 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143862091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systems Metabolic Engineering of Clostridium tyrobutyricum for 1,3‐Propanediol Production From Crude Glycerol","authors":"Yongzhang Mo, Xiaolong Guo, Yang Lan, Jufang Wang, Hongxin Fu","doi":"10.1002/bit.29010","DOIUrl":"https://doi.org/10.1002/bit.29010","url":null,"abstract":"<jats:italic>Clostridium tyrobutyricum</jats:italic> has emerged as a non‐pathogenic microbial cell factory capable of anaerobic production of various value‐added products, such as butyrate, butanol, and butyl butyrate. This study reports the first systematic engineering of <jats:italic>C. tyrobutyricum</jats:italic> for the heterologous production of 1,3‐propanediol (1,3‐PDO) from industrial by‐product crude glycerol. Initially, the glycerol reductive pathway for 1,3‐PDO production was constructed, and the unique glycerol oxidation pathway in <jats:italic>C. tyrobutyricum</jats:italic> was elucidated. Subsequently, the glycerol metabolism and 1,3‐PDO synthesis pathways were enhanced. Furthermore, the intracellular reducing power supply and the fermentation process were optimized to improve 1,3‐PDO production. Consequently, 54.06 g/L 1,3‐PDO with a yield of 0.64 mol/mol and a productivity of 1.13 g/L·h was obtained using crude glycerol and fish meal. The strategies described herein could facilitate the engineering of <jats:italic>C. tyrobutyricum</jats:italic> as a robust host for synthesizing valuable chemicals.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"37 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143853311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Temporal Galactose-Manganese Feeding in Fed-Batch and Perfusion Bioreactors Modulates UDP-Galactose Pools for Enhanced mAb Glycosylation Homogeneity","authors":"Aron Gyorgypal, Erica Fratz-Berilla, Casey Kohnhorst, David N. Powers, Shishir P. S. Chundawat","doi":"10.1002/bit.28999","DOIUrl":"https://doi.org/10.1002/bit.28999","url":null,"abstract":"Monoclonal antibodies (mAbs) represent a majority of biotherapeutics in the market today. These glycoproteins undergo posttranslational modifications, such as N-linked glycosylation, that influence the structural &amp; functional characteristics of the antibody. Glycosylation is a heterogenous posttranslational modification that may influence therapeutic glycoprotein stability and clinical efficacy, which is why it is often considered a critical quality attribute (CQA) of the mAb product. While much is known about the glycosylation pathways of Chinese Hamster Ovary (CHO) cells and how cell culture chemical modifiers may influence the N-glycosylation profile of the final product, this knowledge is often based on the final cumulative glycan profile at the end of the batch process. Building a temporal understanding of N-glycosylation and how mAb glycoform composition responds to real-time changes in the biomanufacturing process will help build integrated process models that may allow for glycosylation control to produce a more homogenous product. Here, we look at the effect of specific nutrient feed media additives (e.g., galactose, manganese) and feeding times on the N-glycosylation pathway to modulate N-glycosylation of a Herceptin biosimilar mAb (i.e., Trastuzumab). We deploy the N-GLYcanyzer process analytical technology (PAT) to monitor glycoforms in near real-time for bench-scale bioprocesses operated in both fed-batch and perfusion modes to build an understanding of how temporal changes in mAb N-glycosylation are dependent on specific media additives. We find that Trastuzumab terminal galactosylation is sensitive to media feeding times and intracellular nucleotide sugar pools. Temporal analysis reveals an increased desirable production of single and double galactose-occupied glycoforms over time under glucose-starved fed-batch cultures. Comparable galactosylation profiles were also observed between fed-batch (nutrient-limited) and perfusion (non-nutrient-limited) bioprocess conditions. In summary, our results demonstrate the utility of real-time monitoring of mAb glycoforms and feeding critical cell culture nutrients under fed-batch and perfusion bioprocessing conditions to produce higher-quality biologics.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"49 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143849358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to “Sequential Assembly of Cell-Laden Hydrogel Constructs to Engineer Vascular-Like Microchannels”","authors":"","doi":"10.1002/bit.29007","DOIUrl":"https://doi.org/10.1002/bit.29007","url":null,"abstract":"<p>Y. Du, M. Ghodousi, H. Qi, N. Haas, W. Xiao, and A. Khademhosseini, “Sequential Assembly of Cell-Laden Hydrogel Constructs to Engineer Vascular-Like Microchannels,” <i>Biotechnology and Bioengineering</i> 108, no. 7 (2011): 1693–1703, 10.1002/bit.23102.</p>\u0000<p>This correction is published due to concerns raised by a third party regarding high similarity between the cell-laden microgels between Figure 4B,D.</p>\u0000<p>The authors explained that the images in Figure 4B,D were from the same sample that was sequentially exposed to different conditions (first exposed to PI and subsequently put in oil) and analyzed. Thus, the microgels appeared repositioned between the imaging sessions. Due to the elapsed time since publication, the original image files were not available, therefore the authors have repeated the experiment in which the gels were not sequentially analyzed to verify the conclusion that “each assembly phase is conducive to cell viability.”</p>\u0000<p>The new data confirmed the same trends as observed before, therefore the experimental results and the corresponding conclusions of the paper remain unaffected.</p>\u0000<p>The corrected Figure 4 is below:</p>\u0000<figure><picture>\u0000<source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/6a30e99e-e16f-4118-ba84-e207a0666764/bit29007-fig-0001-m.jpg\"/><img alt=\"Details are in the caption following the image\" data-lg-src=\"/cms/asset/6a30e99e-e16f-4118-ba84-e207a0666764/bit29007-fig-0001-m.jpg\" loading=\"lazy\" src=\"/cms/asset/2374871b-cf8d-458e-9481-e2c2ee13960c/bit29007-fig-0001-m.png\" title=\"Details are in the caption following the image\"/></picture><figcaption>\u0000<div><strong>Figure 4<span style=\"font-weight:normal\"></span></strong><div>Open in figure viewer<i aria-hidden=\"true\"></i><span>PowerPoint</span></div>\u0000</div>\u0000<div>Viability tests for the cell-laden tubular hydrogels as a function of the steps in the assembly process. The sequential assembly procedure was shown to be cell-friendly as demonstrated by viability tests for: (A) 3T3 fibroblast-laden microgels; (B) microgels washed three times with photoinitiator (PI) (1% in DPBS); (C) microgels exposed to oil; (D) microgels exposed to both PI and oil; (E) microgel assembly after 2 days in culture medium; (F) quantification of samples (<i>n</i> = 3) after each fabrication step showed that the assembly process did not result in a significant loss of cell viability. Scale bar: 500 µm.</div>\u0000</figcaption>\u0000</figure>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"5 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143849528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}