Journal of bioscience and bioengineering最新文献

{"title":"Combined use of phosphorus containing calcium silicate and phosphate-solubilizing bacterium as a solid biofertilizer for sustainable soybean cultivation","authors":"Akiko Hanada ,&nbsp;Tomoaki Kasuga ,&nbsp;Mana Iwamoto ,&nbsp;Akane Mizusawa ,&nbsp;Satoshi Murata ,&nbsp;Tsuyoshi Aketo ,&nbsp;Kazuya Hirata ,&nbsp;Kosuke Kataoka ,&nbsp;Atsushi Arakaki ,&nbsp;Naoko Ohkama-Ohtsu ,&nbsp;Tsuyoshi Tanaka","doi":"10.1016/j.jbiosc.2025.06.008","DOIUrl":"10.1016/j.jbiosc.2025.06.008","url":null,"abstract":"<div><div><span>Recycled phosphorus (P) from wastewater has attracted attention as an alternative P source for sustainable agriculture<span>. We previously developed amorphous calcium silicate<span> hydrate (aCSH) as a simple and efficient material for the recovery of inorganic P from wastewater. Although P recovered using aCSH (P_aCSH) is expected to be useful as a biofertilizer material, research on its effectiveness as a microbial carrier and fertilizer remains unexplored. In this study, we aimed to investigate the potential of the combined use of P_aCSH and a newly isolated phosphate-solubilizing bacterium, </span></span></span><span><span>Burkholderia</span><em> contaminans</em></span><span><span> NKPB12, as a biofertilizer for soybean cultivation. Compared with conventional carriers, P_aCSH showed a higher </span>cell immobilization capacity for NKPB12. Soybean cultivation test results confirmed the promotion of soybean growth when P_aCSH/crystalline CSH (cCSH) with NKPB12 was used as the sole P source. These results indicate that the combination of P_aCSH/cCSH and NKPB12 can be applied as a biofertilizer with recycled P for soybean cultivation. To our knowledge, this is the first report demonstrating the dual functionality of recovered P material as both a P source and bacterial carrier in biofertilizer applications.</span></div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 4","pages":"Pages 211-217"},"PeriodicalIF":2.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144618113","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}

{"title":"Correlation analysis of urinary D/L-amino acid ratios and renal injury by using 5/6 nephrectomy model rats","authors":"Kosuke Fukuda ,&nbsp;Yutaka Umakoshi ,&nbsp;Kenichi Watanabe ,&nbsp;Masahiro Yahata ,&nbsp;Izuru Miyawaki ,&nbsp;Eiichiro Fukusaki","doi":"10.1016/j.jbiosc.2025.06.006","DOIUrl":"10.1016/j.jbiosc.2025.06.006","url":null,"abstract":"<div><div><span>Renal injury, such as </span>chronic kidney disease (CKD), is a global public health problem. Recent studies have reported that several D/L-amino acid ratios are affected during renal injury; however, most reports have focused on renal dysfunction, and few have focused on the association between D/L-amino acid changes and histopathological alterations in the kidney. Therefore, we aimed to investigate the correlation between D/L-amino acid ratios and the histopathological findings of renal injury in a 5/6 nephrectomy rat model. In this study, the D/L-amino acid ratios were evaluated using urine samples collected 1, 2, 3, and 4 weeks after surgery. The D/L-Ser and D/L-Ala ratios were 3.0-fold and 9.3-fold higher, respectively, and the other D/L-amino acid ratios, such as D/L-Arg, D/L-Asn, D/L-His, D/L-Ile, D/L-Lys, D/L-Phe, and D/L-Thr ratios, also showed higher values than those in the sham group. The D/L-amino acid ratios in the urine were consistently maintained throughout the evaluation period compared to those in the sham group. However, only the D/L-Gln ratio was lower than that in the sham group. Correlation analysis between D/L-amino acid ratios and histopathological alterations in the kidney revealed a good correlation between D/L-Ala, D/L-Glu, and D/L-Gln one week after surgery and tubular dilatation. No other findings showed a strong correlation with the changes in the D/L-amino acid ratios of the aforementioned amino acids. By evaluating urinary D/L-amino acid ratios, we could potentially detect tubular dilatation in the kidney noninvasively and obtain insights into the physiological condition of the kidney.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 4","pages":"Pages 260-266"},"PeriodicalIF":2.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144608465","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}

{"title":"Design of a thermostable bilirubin oxidase from Myrothecium verrucaria","authors":"Haruka Kado Horiguchi ,&nbsp;Shohei Yamada ,&nbsp;Hironori Semba ,&nbsp;Hirokazu Tsuboi ,&nbsp;Takayuki Bogaki ,&nbsp;Akio Koda ,&nbsp;Kazuhiko Ishikawa ,&nbsp;Yutaro Mori ,&nbsp;Chiaki Ogino ,&nbsp;Masahiro Takagi ,&nbsp;Yoshio Tsujino","doi":"10.1016/j.jbiosc.2025.05.006","DOIUrl":"10.1016/j.jbiosc.2025.05.006","url":null,"abstract":"<div><div>Bilirubin oxidase (BOD), identified as a multicopper oxidase produced by <em>Myrothecium verrucaria</em>, plays a critical role in the oxidation of bilirubin to biliverdin, which is pivotal in various biochemical processes. To construct a highly thermostable BOD, we have used three protein engineering methods: (i) stabilization of the main chain (proline substitution), (ii) design of salt bridges, and (iii) improvement of hydrophobic interactions. Significant enhancement of thermostability was achieved through stabilization of the main chain (L476P, A496P), introduction of a salt bridge (Q495R), and improvement of hydrophobic interactions (A264V). Furthermore, the combination of these point mutations, which contributed to structural stabilization, resulted in a novel thermostable mutant. Utilizing the cumulative effect of point mutations based on the three strategies, we were able to obtain a thermostable enzyme that exhibited approximately 3.9-fold higher residual activity than wild-type BOD (WT) even after incubation at 60 °C for 1 h and nearly 10 °C higher optimum temperature than that of WT. Importantly, these mutations did not affect its 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) oxidation activity. This approach provides a valuable strategy for improving the thermostability of multivalent copper oxidases and offers promising prospects for industrial applications.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 59-65"},"PeriodicalIF":2.3,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144284503","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}

{"title":"Response to the letter to the editor on “Evaluation methods for decellularized tissues: A focus on human amniotic membrane”","authors":"Miriam Guadalupe Salgado García ,&nbsp;Néstor Fabián Díaz ,&nbsp;Guadalupe García López ,&nbsp;Ikuri Álvarez Maya ,&nbsp;Claudia Hernández Jimenez ,&nbsp;Yvonne Roman Maldonado ,&nbsp;David José Mendoza Aguayo ,&nbsp;Néstor Emmanuel Díaz Martínez","doi":"10.1016/j.jbiosc.2025.05.007","DOIUrl":"10.1016/j.jbiosc.2025.05.007","url":null,"abstract":"","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Page 115"},"PeriodicalIF":2.3,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274956","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}

{"title":"Establishment of a nitrate-reductase promoter-driven inducible expression system for metabolic engineering in the marine diatom Fistulifera solaris","authors":"Tomoki Yamanaka ,&nbsp;Takashi Yabuuchi ,&nbsp;Maeda Yoshiaki ,&nbsp;Tomoko Yoshino ,&nbsp;Kosuke Kataoka ,&nbsp;Tsuyoshi Tanaka","doi":"10.1016/j.jbiosc.2025.05.009","DOIUrl":"10.1016/j.jbiosc.2025.05.009","url":null,"abstract":"<div><div>Microalgae have emerged as promising hosts for producing valuable compounds, including lipids and recombinant proteins. The oleaginous diatom <em>Fistulifera solaris</em> has shown potential for industrial applications. However, the constitutive expression of proteins often causes cellular toxicity, necessitating a controlled gene expression system to enhance productivity. In this study, we characterized the endogenous promoter of the nitrate reductase (NR) gene as an inducible expression system. The promoter was activated by nitrate and suppressed by ammonium. We also established a nitrate-spike method for controlled promoter activation by adding NaNO₃ at specific time points, which achieved strong induction without compromising the cellular lipid content. This study provides the first characterization of the NR promoter in an oleaginous diatom and demonstrates its use for controlled compound production. The inducible expression system developed in this study represents fundamental technology for enhancing the productivity of <em>F. solaris</em>.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 73-78"},"PeriodicalIF":2.3,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144248068","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}

{"title":"Recovery of hyaluronic acid using thermosensitive polymer-salt aqueous biphasic system with polymer recycling","authors":"Nagaraju Marimuthu ,&nbsp;Yin Hui Chow ,&nbsp;Phei Er Kee ,&nbsp;John Chi-Wei Lan ,&nbsp;Zee Wei Lai ,&nbsp;Li Wan Yoon ,&nbsp;Yew Joon Tam","doi":"10.1016/j.jbiosc.2025.04.007","DOIUrl":"10.1016/j.jbiosc.2025.04.007","url":null,"abstract":"<div><div>Microbial fermentation emerges as a viable alternative to animal tissue-based extraction for industrial hyaluronic acid (HA) production. However, microbial HA requires rigorous separation and purification processes to meet high purity standards for cosmeceutical use. Aqueous biphasic system (ABS) presents a promising strategy for HA recovery due to its cost-effectiveness, high yield, purity and eco-friendly nature. Thermosensitive polymer, specifically ethylene oxide-propylene oxide (EOPO)-salt ABS, was proposed in the present study to recover microbial HA from <em>Streptococcus zooepidemicus</em> fermentation broth. The effects of molecular weight (MW) of EOPO, types of salt, phase composition, and the amount of crude extract on the efficiency of ABS for HA recovery were studied. Furthermore, the recovery efficiency of ABS formed using recycled EOPO was investigated. HA exhibited a preference for the salt-rich bottom phase of ABS because of its hydrophilic surface features. Highest partition coefficient of 11.41 ± 0.00 and yield of 93.42 % ± 0.00 % were achieved in ABS containing 14 % (w/w) EOPO₂₅₀₀, 12 % (w/w) sodium citrate at pH 8.2 and 15 % (w/w) crude extract. More than 70 % of EOPO₂₅₀₀ was recovered through the thermo-separation process, and the 1st recycling of EOPO₂₅₀₀ maintained a recovery yield of HA above 80 % in ABS. Moreover, Fourier transform infrared spectroscopy validated that HA retained its structure after the recovery process. In conclusion, this study demonstrates the effectiveness of EOPO-salt ABS for microbial HA recovery, achieving high recovery yield. The recycling of EOPO enhances the economic viability and environmental sustainability of the system. These findings pave the way for greener, scalable HA production practices while maintaining product integrity.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 79-89"},"PeriodicalIF":2.3,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144248069","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}

{"title":"Discovery and affinity maturation of antibody fragments from an unfavorably enriched phage display selection by deep sequencing and machine learning","authors":"Sakiya Kawada ,&nbsp;Yoichi Kurumida ,&nbsp;Tomoyuki Ito ,&nbsp;Thuy Duong Nguyen ,&nbsp;Hafumi Nishi ,&nbsp;Hikaru Nakazawa ,&nbsp;Yutaka Saito ,&nbsp;Tomoshi Kameda ,&nbsp;Koji Tsuda ,&nbsp;Mitsuo Umetsu","doi":"10.1016/j.jbiosc.2025.05.004","DOIUrl":"10.1016/j.jbiosc.2025.05.004","url":null,"abstract":"<div><div>Phage display selection has been used for directed evolution of antibody fragments. However, variants with binding affinity cannot be always identified due to undesirable enrichment of target-unrelated variants in the biopanning process. Here, our goal was to obtain functional variants by deep sequencing and machine learning from a phage display library where functional variants were not appropriately enriched. Deep sequencing of the previously biopanned pools revealed that amplification bias might have prevented the enrichment of target-binding phages. We performed a sequence similarity search based on the deep sequencing analysis so that the influence of bias was decreased, leading to discovery of a variant with binding affinity, which could not be discovered by a conventional screening method alone. We applied machine learning to the deep sequencing data; the machine learning proposed effective mutations for increasing affinity, allowing us to identify a variant with improved affinity (EC₅₀ = 3.46 μM). In summary, we present the possibility of obtaining functional variants even from unfavorably enriched phage libraries by using deep sequencing and machine learning.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 51-58"},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144234236","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}

{"title":"Preparation of minicircle DNA vectors for gene knock-in to mammalian cells","authors":"Yoshinori Kawabe,&nbsp;Makoto Hamaoka,&nbsp;Akio Kuno,&nbsp;Yusaku Yoshimura,&nbsp;Masamichi Kamihira","doi":"10.1016/j.jbiosc.2025.05.005","DOIUrl":"10.1016/j.jbiosc.2025.05.005","url":null,"abstract":"<div><div>Minicircle DNA vectors (MCs) are gene carriers with favorable characteristics for gene therapy and the production of recombinant cells using animal cells as hosts. Typically, MCs are prepared by inserting target gene fragments as an expression unit into a standard plasmid vector, followed by the removal of bacterial sequences. In this study, we report a method for MC preparation utilizing the Cre-<em>loxP</em> recombination system. By inserting the target gene fragments between two <em>loxP</em> sites introduced into the plasmid vector, the plasmid backbone can be removed through the action of Cre recombinase. We explored optimal reaction conditions for MC generation using Cre. Furthermore, site-specific knock-ins into the CHO cell genome were performed using the remaining <em>loxP</em> sequence in the generated MCs via the Cre-<em>loxP</em> reaction. When MCs were used as gene donors, significant improvements in gene integration efficiency and enhanced gene expression in CHO cells were observed compared with conventional plasmids. The MC preparation and gene knock-in techniques developed in this study are highly useful for CHO cell engineering.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 107-112"},"PeriodicalIF":2.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187060","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}

{"title":"Construction of contractile human iPSC-derived skeletal muscle tissues on 96-well scale microdevices","authors":"Seitaro Nakamura ,&nbsp;Yuhei Kamei ,&nbsp;Ayumu Matsushima ,&nbsp;Kazuki Yamamoto ,&nbsp;Hirokazu Akiyama ,&nbsp;Muhammad Irfanur Rashid ,&nbsp;Yohei Okada ,&nbsp;Tomoya Uchimura ,&nbsp;Hidetoshi Sakurai ,&nbsp;Hiroyuki Honda ,&nbsp;Kazunori Shimizu","doi":"10.1016/j.jbiosc.2025.05.003","DOIUrl":"10.1016/j.jbiosc.2025.05.003","url":null,"abstract":"<div><div>Tissue-engineered three-dimensional (3D) skeletal muscles can be potentially used in contractile force-based phenotypic screening to elucidate the mechanisms of skeletal muscle dysfunction and develop preventive and therapeutic strategies. Human induced pluripotent stem cells (hiPSCs) expressing tetracycline-inducible myogenic differentiation 1 (MYOD1) are a promising cell source for construction of tissue-engineered skeletal muscles. Although we successfully constructed contractile tissues using these hiPSCs in a previous study, further improvements are required because of their weak contractile force and inefficient screening capabilities. In this study, we aimed to construct iPSC-derived muscle tissues with high contractile force using a 96-well scale microdevice that we had previously developed. To increase the contractile force, we optimized the time of supplementation of the transforming growth factor-β (TGF-β) inhibitor, SB431542 (SB), to identify culture conditions that enhance contractile force. The maximum contractile force with addition of SB was approximately five times greater than that without SB (58.45 ± 20.14 μN with SB compared to 11.64 ± 4.86 μN without SB). Various analyses, including immunostaining, transmission electron microscopy, gene expression analysis, and proteomics, revealed enhanced myotube differentiation and muscle tissue maturation in the presence of SB. Experiments using inhibitors indicated that TGFβ1, not myostatin, is partially involved in these effects. Furthermore, we confirmed that tissues constructed from iPSCs derived from patients with Duchenne muscular dystrophy also showed improved contractility following addition of SB. Therefore, iPSC-derived muscle tissues cultured with SB on the 96-well microdevice provide a promising platform for screening compounds that can ameliorate disease pathology.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 98-106"},"PeriodicalIF":2.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181801","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}

{"title":"Novel promoter of Pseudozyma antarctica exhibits stable glucose-mediated activation and copper-mediated suppression of flanking genes","authors":"Tohru Yarimizu ,&nbsp;Atsuhiro Miura ,&nbsp;Xiao-hong Cao ,&nbsp;Takumi Tanaka ,&nbsp;Tomotake Morita ,&nbsp;Hitoshi Shimoi ,&nbsp;Hirokazu Ueda ,&nbsp;Hiroko Kitamoto","doi":"10.1016/j.jbiosc.2025.05.002","DOIUrl":"10.1016/j.jbiosc.2025.05.002","url":null,"abstract":"<div><div><em>Pseudozyma antarctica</em> is a leaf-colonizing basidiomycetous yeast that produces a plastic-degrading enzyme (PaE) using xylose as an inducer and carbon source. Notably, glucose, a readily available carbon source, has not been utilized for PaE production. Herein, we identified two promoters of <em>P. antarctica</em> for stable induction of PaE gene transcription at high levels in commonly used yeast media with glucose, along with assessing the effect of copper supplementation. Through microarray analysis, the top five genes with high expression in glucose cultures were identified and their promoter activities were evaluated by reporter assay. The first and second genes were located 903 nucleotides apart on the <em>P. antarctica</em> chromosome and shared reciprocal inverted sequences as promoters, namely Pa<em>FRE1</em> and Pa<em>CTR1</em>, respectively, owing to their weak similarity to <em>FRE</em> and <em>CTR</em> in <em>Saccharomyces cerevisiae</em> genome, which regulate the copper uptake. When <em>P. antarctica</em> was grown in three common media using glucose as a carbon source, both Pa<em>FRE1</em> and Pa<em>CTR1</em> induced high and stable expression and produced more heterologous artificial luciferase and endogenous PaE than that by the highly expressed actin promoter. Additionally, the activities of Pa<em>FRE1</em> and Pa<em>CTR1</em> promoters were suppressed by copper addition. Altogether, this study shows that these <em>P. antarctica</em> promoters can be used to produce various proteins such as PaE.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 2","pages":"Pages 66-72"},"PeriodicalIF":2.3,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181644","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}