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A high-resolution transcriptomic atlas depicting nitrogen fixation and nodule development in soybean

Journal of Integrative Plant Biology 2023年第6期65卷 1536-1552页

作者： Baocheng Sun Yu Wang Qun Yang han gao Haiyu Niu Yansong Li Qun Ma Qing Huan Wenfeng Qian Bo Ren State Key Laboratory of Plant Genomics Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China

Although root nodules are essential for biological nitrogen fixation in legumes,the cell types and molecular regulatory mechanisms contributing to nodule development and nitrogen fixation in determinate nodule legumes,such as soybean(Glycine max),remain incompletely understood.Here,we generated a single-nucleus resolution transcriptomic atlas of soybean roots and nodules at 14 days post inoculation(dpi)and annotated 17 major cell types,including six that are specific to nodules.We identified the specific cell types responsible for each step in the ureides synthesis pathway,which enables spatial compartmentalization of biochemical reactions during soybean nitrogen fixation.By utilizing RNA velocity analysis,we reconstructed the differentiation dynamics of soybean nodules,which differs from those of indeterminate nodules in Medicago truncatula.Moreover,we identified several putative regulators of soybean nodulation and two of these genes,GmbHLH93 and GmSCL1,were as-yet uncharacterized in soybean.Overexpression of each gene in soybean hairy root systems validated their respective roles in nodulation.Notably,enrichment for cytokinin-related genes in soybean nodules led to identification of the cytokinin receptor,GmCRE1,as a prominent component of the nodulation pathway.GmCRE1 knockout in soybean resulted in a striking nodule phenotype with decreased nitrogen fixation zone and depletion of leghemoglobins,accompanied by downregulation of nodule-specific gene expression,as well as almost complete abrogation of biological nitrogen fixation.In summary,this study provides a comprehensive perspective of the cellular landscape during soybean nodulation,shedding light on the underlying metabolic and developmental mechanisms of soybean nodule formation.

关键词： cytokinin determinate nodule nitrogen fixation nodulation RNA velocity single nucleus RNA sequencing(snRNA-seq) soybean ureide

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RR-1 cuticular protein TcCPR69 is required for growth and metamorphosis in Tribolium castaneum

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Insect Science 2022年第6期29卷 1612-1628页

作者： Jia Xie Guifang Peng Miao Wang Qisheng Zhong Xiaowen Song Jingxiu Bi Jing Tang Fan Feng han gao Bin Li Jiangsu Key Laboratory of Biodiversity and Biotechnology College of Life SciencesNanjing Normal UniversityNanjingChina Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests Ministry of EducationCollege of Plant Protection HainanUniversityHaikouChina

Cuticle is not only critical for protecting insects from noxious stimuli but is also involved in a variety of metabolic activities.Cuticular proteins(CPs)affect cuticle structure and mechanical properties during insect growth,reproduction,and environmental adaptation.Here,we describe the identification and characterization of a member of the RR-1 subfamily of CPs with an R&R consensus(CPR)in Tribolium castaneum(TcCPR69).Although it was previously reported to be highly expressed in the wings,we found that knocking down TcCPR69 by RNA interference(RNAi)did not cause obvious wing abnormalities but markedly disrupted the growth and metamorphosis of beetles with 100%cumulative mortality;additionally,the chitin content of the pharate adult was decreased and the new abdominal cuticle was significantly thinner before molting.TcCPR69 showed chitin-binding ability and the expression levels of key genes involved in chitin metabolism(trehalase[TcTRE],chitin synthase[TcCHSA and TcCHSB],and chitinase[TcCHT5 and TcCHT10])were also decreased by TcCPR69 knockdown.TcCPR69 gene expression peaked shortly after molting and was increased 2.61 fold at 12 h after 20-hydroxyecdysone(20E)injection.This was reversed by RNAi of the ecdysone-related genes ecdysone receptor(TcECR)and fushi tarazu transcription factor 1(TcFTZ-F1).These results indicate that TcCPR69 is positively regulated by 20E signaling to contribute to cuticle formation and maintain chitin accumulation during the growth and metamorphosis of beetles.

关键词： chitin accumulation cuticle protein development 20-hydroxyecdysone molting RNA interference

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Effects of polyol excipient stability during storage and use on the quality of biopharmaceutical formulations

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Journal of Pharmaceutical Analysis 2022年第5期12卷 774-782页

作者： Min-Fei Sun Jia-Ning Liao Zhen-Yi Jing han gao Bin-Bin Shen You-Fu Xu Wei-Jie Fang Institute of Drug Metabolism and Pharmaceutical Analysis College of Pharmaceutical SciencesZhejiang UniversityHangzhou310058China hangzhou Institute of Innovative Medicine Zhejiang UniversityHangzhou Biopharm TownHangzhou310016China Zhejiang Haisheng Pharmaceutical Inc. TaizhouZhejiang318000China

Biopharmaceuticals are formulated using a variety of excipients to maintain their storage stability.However,some excipients are prone to degradation during repeated use and/or improper storage,and the impurities generated by their degradation are easily overlooked by end users and are usually not strictly monitored,affecting the stability of biopharmaceuticals.In this study,we evaluated the degradation profile of polyol excipient glycerol during repeated use and improper storage and identified an unprecedented cyclic ketal impurity using gas chromatography with mass spectrometry(GC-MS).The other polyol excipient,mannitol,was much more stable than glycerol.The effects of degraded glycerol and mannitol on the stability of the model biopharmaceutical pentapeptide,thymopentin,were also evaluated.The thymopentin content was only 66.4% in the thymopentin formulations with degraded glycerol,compared to 95.8% in other formulations after the stress test.Most glycerol impurities(i.e.,aldehydes and ketones)reacted with thymopentin,affecting the stability of thymopentin formulations.In conclusion,this work suggests that more attention should be paid to the quality changes of excipients during repeated use and storage.Additional testing of excipient stability under real or accelerated conditions by manufacturers would help avoid unexpected and painful results.

关键词： Excipient stability GC-MS Glycerol LC-MS/MS Mannitol Thymopentin

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Characterization,heterologous expression and engineering of trehalase for biotechnological applications

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Systems Microbiology and Biomanufacturing 2022年第3期2卷 445-460页

作者： han gao Jin-Song Gong Chang Su Heng Li Zheng-Hong Xu Jin-Song Shi Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of EducationSchool of Life Sciences and Health EngineeringJiangnan UniversityLihu Avenue No.1800Wuxi 214122People’s Republic of China National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing School of BiotechnologyJiangnan UniversityWuxi 214122People’s Republic of China Jiangsu Provincial Engineering Research Center for Bioactive Product Processing Jiangnan UniversityWuxi 214122People’s Republic of China

Trehalose is a non-reducing disaccharide connected byα-1,1-glycosidic bonds;it is widely distributed in bacteria,fungi,yeast,insects,and plant tissues and plays various roles.It can be hydrolyzed by trehalase into two glucose molecules.Trehalases from different sources have been expressed in Escherichia coli,Pichia pastoris,Saccharomyces cerevisiae,baculovirus-silkworm,and other expression systems;however,it is most common in E.coli.The structural characteristics of different glycoside hydrolase(GH)family trehalases and the sources of trehalase have been analyzed.The catalytic mechanism of GH37 trehalase has also been elucidated in detail.Moreover,the molecular modification of trehalase has mainly focused on directed evolution for improving enzyme activity.We comprehensively reviewed the current application status and adaptable transformations was comprehensively overviewed in the context of industrial performance.We suggest that the level of recombinant production is far from meeting industrial requirements,and the catalytic performance of trehalase needs to be improved urgently.Finally,we discuss developmental prospects and future trends.

关键词： Trehalase Structure Expression Catalytic mechanism Application

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Anti-glial fibrillary acidic protein antibody and anti-aquaporin-4 antibody double-positive neuromyelitis optica spectrum disorder:A case report

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World Journal of Clinical Cases 2023年第34期11卷 8192-8199页

作者： Ting-Yu Jin Bing-Tong Lin Li-Jv Dai Xia Lu han gao Jin Hu Department of Neurology First Affiliated Hospital of Jiaxing UniversityJiaxing 314000Zhejiang ProvinceChina

BACKGROUND A case of neuromyelitis optica spectrum disorder(NMOSD)with positive cerebrospinal fluid(CSF)anti-aquaporin-4 antibody(AQP4-IgG)and anti-glial fibrillary acidic protein IgG(GFAP-IgG)at the time of relapse was reported.The exact roles of GFAP-IgG in NMOSD are not fully understood and are the subject of ongoing research.This study revealed the possible connection between GFAPIgG and the occurrence or development of diseases.CASE SUMMARY A 19-year-old woman was admitted to the hospital due to a constellation of symptoms,including dizziness,nausea,and vomiting that commenced 1 year prior,reoccurred 2 mo ago,and were accompanied by visual blurring that also began 2 mo ago.Additionally,she presented with slurred speech and ptosis,both of which emerged 1 mo ago.Notably,her symptoms deteriorated 10 d prior to admission,leading to the onset of arm and leg weakness.During hospitalization,magnetic resonance imaging showed high T2-fluid attenuated inversion recovery signals,and slightly high and equal diffusion-weighted imaging signals.The serum antibody of AQP4-IgG tested positive at a dilution of 1:100.CSF antibody testing showed positive results for GFAP-IgG at a dilution of 1:10 and AQP4-IgG at a dilution of 1:32.Based on these findings,the patient was diagnosed with NMOSD.She received intravenous methylprednisolone at a daily dose of 500 mg for 5 d,followed by a tapering-off period.Afterward,the rate of reduction was gradually slowed down and the timely use of immunosuppressants was implemented.CONCLUSION The CFS was slightly GFAP-IgG-positive during the relapse period,which can aid in the diagnosis and treatment of the disease.

关键词： Anti-glial fibrillary acidic protein antibody Neuromyelitis optica spectrum disorder Anti-aquaporin-4 antibody Cerebrospinal fluid Case report

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Development and validation of a novel criterion of histologic healing in ulcerative colitis defined by inflammatory cell enumeration in lamina propria mucosa: A multicenter retrospective cohort in China

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中华医学杂志(英文版) 2024年第11期137卷 2024 May 13页

作者： gao han Peng Kangsheng Shi Yadi Zhu Shenshen Sun Ruicong Xu Chunjin Liu Ping Pang Zhi Zhu Lanxiang Chen Weichang Feng Baisui Wu Huili Zhou Guangxi Li Mingsong Li Junxiang Ding Baijing Liu Zhanju Department of Gastroenterology The Third Affiliated Hospital of Guangzhou Medical University Guangzhou Guangdong China Department of Gastroenterology Dongfang Hospital of Beijing University of Chinese Medicine Beijing China Center for IBD Research and Department of Gastroenterology Shanghai Tenth People’s Hospital Tongji University School of Medicine Shanghai China Clinical Medicine Sanquan College of Xinxiang Medical University Xinxiang Henan China Department of Gastroenterology The First People’s Hospital of Shangqiu City Affiliated to Xinxiang Medical University Shangqiu Henan China Department of Gastroenterology Wuhu First People’s Hospital Wuhu Anhui China Department of Gastroenterology Suzhou Municipal Hospital Affiliated to Nanjing Medical University Suzhou Jiangsu China Department of Gastroenterology The First Affiliated Hospital of Soochow University Suzhou Jiangsu China Department of Gastroenterology The Second Affiliated Hospital of Zhengzhou University Zhengzhou Henan China Department of Gastroenterology Zhengzhou Central Hospital Affiliated to Zhengzhou University Zhengzhou Henan China Department of Gastroenterology The Affiliated Hospital of Jining Medical College Jining Shandong China

Background: Histological healing is closely associated with improved long-term clinical outcomes and lowered relapses in patients with ulcerative colitis (UC). Here, we developed a novel diagnostic criterion for assessing histological healing in UC patients.Methods: We conducted a retrospective cohort study in UC patients, whose treatment was iteratively optimized to achieve mucosal healing at Shanghai Tenth People’s Hospital of Tongji University from January 2017 to May 2022. We identified an inflammatory cell enumeration index (ICEI) for assessing histological healing based on the proportions of eosinophils, CD177+ neutrophils, and CD40L+ T cells in the colonic lamina propria under high power field (HPF), and the outcomes (risks of symptomatic relapses) of achieving histological remissionvs. persistent histological inflammation using Kaplan-Meier curves. Intrareader reliability and inter-reader reliability were evaluated by each reader. The relationships to the changes in the Nancy index and the Geboes score were also assessed for responsiveness. The ICEI was further validated in a new cohort of UC patients from other nine university hospitals.Results: We developed an ICEI for clinical diagnosis of histological healing, i.e., Y = 1.701X1 + 0.758X2 + 1.347X3 - 7.745 (X1, X2, and X3 represent the proportions of CD177+ neutrophils, eosinophils, and CD40L+ T cells, respectively, in the colonic lamina propria under HPF). The receiver operating characteristics curve (ROC) analysis revealed that Y <-0.391 was the cutoff value for the diagnosis of histological healing and that an area under the curve (AUC) was 0.942 (95% confidence interval [CI]: 0.905-0.979) with a sensitivity of 92.5% and a specificity of 83.6% (P <0.001). The intraclass correlation coefficient (ICC) for the intrareader reliability was 0.855 (95% CI: 0.781-0.909), and ICEI had good inter-reader reliability of 0.832 (95% CI: 0.748-0.894). During an 18-month follow-up, patients with histological healing had a substantially better outcome compared with

关键词： Ulcerative colitis Histological healing Inflammatory cell enumeration index Prognosis

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Modulating reaction pathways of formic acid oxidation for optimized electrocatalytic performance of PtAu/CoNC

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Nano Research 2022年第2期15卷 1221-1229页

作者： Mengchao Liang Tianyu Xia han gao Kai Zhao Tianqi Cao Meng Deng Xiaoyan Ren Shunfang Li Haizhong Guo Rongming Wang Key Laboratory of Material Physics Ministry of EducationSchool of Physics and MicroelectronicsZhengzhou UniversityZhengzhou 450052China Collaborative Innovation Center of Light Manipulations and Applications Shandong Normal UniversityJinanShandong 250358China Beijing Advanced Innovation Center for Materials Genome Engineering Beijing Key Laboratory for Magneto-Photoelectrical Composite and Interface ScienceSchool of Mathematics and PhysicsUniversity of Science and Technology BeijingBeijing 100083China

Formic acid oxidation(FAO)is a typical anode reaction in fuel cells that can be facilitated by modulating its direct and indirect reaction pathways.Herein,PtAu bimetallic nanoparticles loaded onto Co and N co-doping carbon nanoframes(CoNC NFs)were designed to improve the selectivity of the direct reaction pathway for efficient FAO.Based on these subtle nanomaterials,the influences of elemental composition and carbon-support materials on the two pathways of FAO were investigated in detail.The results of fuel cell tests verified that the appropriate amount of Au in PtAu/CoNC can promote a direct reaction pathway for FAO,which is crucial for enhancing the oxidation efficiency of formic acid.In particular,the obtained PtAu/CoNC with an optimal Pt/Au atomic ratio of 1:1(PtAu/CoNC-3)manifests the best catalytic performance among the analogous obtained Pt-based electrocatalysts.The FAO mass activity of the PtAu/CoNC-3 sample reached 0.88 A·mg_(Pt)^(-1),which is 26.0 times higher than that of Pt/C.The results of first-principles calculation and CO stripping jointly demonstrate that the CO adsorption of PtAu/CoNC is considerably lower than that of Pt/CoNC and PtAu/C,which indicates that the synergistic effect of Pt,Au,and CoNC NFs is critical for the resistance of Pt to CO poisoning.This work is of great significance for a deeper understanding of the oxidation mechanism of formic acid and provides a feasible and promising strategy for enhancing the catalytic performance of the catalyst by improving the direct reaction pathway for FAO.

关键词： formic acid oxidation direct oxidation pathway metal organic framework co-doping CO binding energy

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Reading Time and DNA Sequence Preference of TET3 CXXC Domain Revealed by Single-Molecule Profiling

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Chinese Journal of Chemistry 2023年第10期41卷 1177-1184页

作者： Zeyu Wang Zhiqiang Cao Kangkang Ma Man Lu Meije Wang han gao Deshun Gong Lin Liang Zhongbo Yu Frontiers Science Center for Cell Responses State Key Laboratory of Medicinal Chemical Biologyand College of PharmacyNankai University38 Tongyan RoadTianjin 300350China State Key Laboratory of Medicinal Chemical Biology and College of Life Sciences Nankai UniversityTianjin 300350China College of Animal Science Shanxi Agricultural UniversityTaiguShanxi 030801China

Recognition of CpG dinucleotide DNA in epigenetic information flow plays a pivotal role for cellular differentiation and development.The TET3 CXXC domain binds to CpG DNA,serving a basic epigenetic information reading mechanism.During the selective recognition of a CpG motif by a CXXC domain from crowded binding sites in a gene sequence,the protein-DNA interactions are beyond CpG dinu-cleotide.However,the selective binding dynamics of CpG within a long DNA context by epigenetic enzymes have been rarely exploit-ed,which is hard for ensemble methods to probe.Here,we used single-molecule magnetic tweezers to quantitatively examine the dynamics of TET3's CXXC domain on a Hoxa9 promoter DNA.Our single-molecule binding profile revealed that CXXC-DNA interactions involve both CpG motifs and their flanking sequences.The residence time of TET3 CXXC differs by about 1000 times in five distin-guished CpG clusters in the context of a CpG island.Moreover,we performed multi-state hidden Markov modeling analysis on the zip-ping/unzipping dynamics of a CpG hairpin,discovering TET3 CXXC's preference on CpG motifs regarding the-2 to+2 flanking bases.Our results shed light on the selective binding dynamics of a CXXC on a gene sequence,facilitating studies on epigenetic information reading mechanisms.

关键词： Enzymes DNA recognition CpG flanking sequence Single-molecule studies Magnetic tweezers

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Research Trends of Macrophage Polarization:A Bibliometric Analysis

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Chinese Medical Journal 2018年第24期131卷 2968-2975页

作者： han gao Feng-Yan Huang Zhi-Ping Wang Department of Occupational and Environmental Health School of Public HealthShandong UniversityJinanShandong 250012China

Background:Macrophage polarization is involved in the development of many diseases such as obesity,diabetes,and cancer.This study aimed to understand the trends and hotspots ofmacrophage polarization research. Methods:We searched through the Web of Science Core Collection database to obtain original articles in this research domain.CiteSpace, HistCite,and VOSviewer software were used to facilitate the analysis and visualization of scientific productivity and emerging trends. Results:The survey included 3064 articles,and the annual number of publications exhibited an exponential increase.These articles have received a total of 74,801 citations,and the number of annual citations grew from 68 to 18,074 in a decade.Research on macrophage polarization was performed in 76 countries,and the USA ranked first in terms of research output by contributing 1129 (36.8%)articles. The USA also had the highest H-index,total citations,and highly cited article number.PLOS One,Journal of Immunology,and Scientific Reports were the three journals that published the most articles.Interdisciplinary research areas involving macrophage polarization,such as biomaterials,cancer,and diabetes,were identified by journal citation analysis.The top 20 most productive institutions were located mainly in the USA,France,and China,and top authors originated mainly from the USA and Italy.Tumor biology,obesity,and infection were research hotspots and may be promising in the next few years. Conclusions:This study provides a comprehensive analysis that delineates the scientific productivity,collaboration,and research hotspots of macrophage polarization research.

关键词： Bibliometric Analysis Macrophage Macrophage Polarization

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The latest breakthrough on genetic characteristics ofinflammatory bowel disease in Chinese and other EastAsianancestries

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Precision Clinical Medicine 2023年第3期6卷 135-140页

作者： han gao Zhanju Liu Center for Inflammatory Bowel Disease Research and Department of Gastroenterology The Shanghai Tenth People's HospitalTongji UniversityShanghai 200072China

Inflammatory bowel diseases(IBDs)are complex chronic disorders of the gastrointestinal tract with the following two subtypes:Crohn's disease and ulcerative colitis.Disease presentation and progression within and across IBDs,especially Crohn's disease,are highly heterogeneous in the location,severity of inflammation,intestinal stenosis and obstruction,and extraintestinal manifestations.Clinical classifications fail to accurately predict the disease course and response to therapies.To date,most IBD genetic associations are derived from individuals of European ancestries,leading to a limitation of the discovery and application of IBD genetics in the rest of the world populations.In this mini-review,we summarize the latest progress of genome-wide association studies of IBD across global ancestries especially the Chinese population,the similarities and differences in genetic architecture between European and East Asian ancestries,as well as,the clinical significances relevant to IBD genetic study.

关键词： Chinese population Crohn's disease East Asian infammatory bowel disease susceptible gene ulcerative colitis

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