Journal Article
- Fresh Washed Microbiota Transplantation Alters Gut Microbiota Metabolites to Ameliorate Sleeping Disorder Symptom of Autistic Children
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Nai-Hua Liu , Hong-Qian Liu , Jia-Yi Zheng , Meng-Lu Zhu , Li-Hao Wu , Hua-Feng Pan , Xing-Xiang He
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J. Microbiol. 2023;61(8):741-753. Published online September 4, 2023
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DOI: https://doi.org/10.1007/s12275-023-00069-x
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Accumulating studies have raised concerns about gut dysbiosis associating autism spectrum disorder (ASD) and its related
symptoms. However, the effect of gut microbiota modification on the Chinese ASD population and its underlying mechanism
were still elusive. Herein, we enrolled 24 ASD children to perform the first course of fresh washed microbiota transplantation
(WMT), 18 patients decided to participate the second course, 13 of which stayed to participate the third course, and there were
8 patients at the fourth course. Then we evaluated the effects of fresh WMT on these patients and their related symptoms.
Our results found that the sleeping disorder symptom was positively interrelated to ASD, fresh WMT significantly alleviated
ASD and its sleeping disorder and constipation symptoms. In addition, WMT stably and continuously downregulated Bacteroides/
Flavonifractor/Parasutterella while upregulated Prevotella_9 to decrease toxic metabolic production and improve
detoxification by regulating glycolysis/myo-inositol/D-glucuronide/D-glucarate degradation, L-1,2-propanediol degradation,
fatty acid β-oxidation. Thus, our results suggested that fresh WMT moderated gut microbiome to improve the behavioral
and sleeping disorder symptoms of ASD via decrease toxic metabolic production and improve detoxification. Which thus
provides a promising gut ecological strategy for ASD children and its related symptoms treatments.
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Citations
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- Effects of gastrointestinal symptoms on the efficacy of washed microbiota transplantation in patients with autism
Dong-Xia Hu, Cai-Mei Lu, Xin-Yu Si, Qing-Ting Wu, Li-Hao Wu, Hao-Jie Zhong, Xing-Xiang He
Frontiers in Pediatrics.2025;[Epub] CrossRef - The Impact of Fecal Microbiota Transplantation on Gastrointestinal and Behavioral Symptoms in Children and Adolescents with Autism Spectrum Disorder: A Systematic Review
Anna Liber, Małgorzata Więch
Nutrients.2025; 17(13): 2250. CrossRef - Pathogenic bacteria enriched in the oral microbiota might be associated with recurrent pulmonary infections in elderly individuals
Jingyi Xu, Ruyi Qu, Keke Yang, Yuezhu Wang, Meiyun Nie, Xiaodong Qi, Huajun Zheng, Ling Yang
Aging Clinical and Experimental Research.2025;[Epub] CrossRef - Does constipation affect the effectiveness of washed microbiota transplantation in treating autism spectrum disorders?
Zihao Pan, Zheng Gao, Junyi Chen, Yongxi Quan, Jiating Xu, Xiaofeng Liang, Wenrui Xie, Xingxiang He, Lihao Wu
Frontiers in Neuroscience.2025;[Epub] CrossRef - Untargeted urine metabolomics and machine learning provide potential metabolic signatures in children with autism spectrum disorder
Xian Liu, Xin Sun, Cheng Guo, Zhi-Fang Huang, Yi-Ru Chen, Fang-Mei Feng, Li-Jie Wu, Wen-Xiong Chen
Frontiers in Psychiatry.2024;[Epub] CrossRef - Washed Microbiota Transplantation Improves the Sleep Quality in Patients with Inflammatory Bowel Disease
Qianqian Li, Yujie Liu, Zulun Zhang, Sheng Zhang, Xiao Ding, Faming Zhang
Nature and Science of Sleep.2024; Volume 16: 1141. CrossRef
Research Support, Non-U.S. Gov'ts
- Expression, Purification, and Characterization of Recombinant Fibulin-5 in a Prokaryote Expression System
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Myoung Seok Jeong , Chang Soo Kang , Yeon Soo Han , In Seok Bang
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J. Microbiol. 2010;48(5):695-700. Published online November 3, 2010
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DOI: https://doi.org/10.1007/s12275-010-0320-6
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189
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Fibulin-5 is a widely expressed, integrin-binding extracellular matrix protein that mediates endothelial cell adhesion and scaffolds cells to elastic fibers. To investigate anti-angiogenesis activities and context-specific activities on responsive cells of recombinant fibulin-5 (rfibulin-5) expressed in Escherichia coli, the cDNA of fibulin-5 cloned from a human placenta cDNA library was inserted into the pET32a (+) vector to allow fibulin-5 expression as a Trx fusion protein. The fusion protein Trx-fibulin-5, expressed as insoluble inclusion bodies, was solubilized and its resulting expression level reached to 15% of the total cell protein. The Trxfibulin-5 was purified effectively by N2+-chelating chromatography and then identified by Western blotting analysis with an anti-His tag antibody. The purified Trx-fibulin-5 was refolded by dialysis against redox reagents, and the rfibulin-5 released from the fusion protein by enterokinase cleavage was purified using a RESOURCE RPC column. The final purified rfibulin-5 effectively inhibited angiogenesis in chicken embryos in a dose-dependent manner through a chorioallantoic membrane (CAM) assay. Additionally, rfibulin-5 potently suppressed in vitro proliferation of human umbilical vein endothelial cells, but stimulated that of human dermal fibroblasts. The expression and in vitro refolding of rfibulin-5 resulted in production of an active molecule with a yield of 2.1 mg/L.
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Citations
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- Impact of UV pre-treatment on the Longissimus thoracis et lumborum muscle proteomes of dry-aged beef cuts: A characterisation within two sampling locations
Sara Álvarez, Carlos Álvarez, Anne Maria Mullen, Eileen O'Neill, Mohammed Gagaoua
Meat Science.2025; 221: 109729. CrossRef - Genome-wide association study on reproductive traits in Jinghai Yellow Chicken
G.X. Zhang, Q.C. Fan, J.Y. Wang, T. Zhang, Q. Xue, H.Q. Shi
Animal Reproduction Science.2015; 163: 30. CrossRef - Production and on-column re-folding of human vascular endothelial growth factor 165 in Escherichia coli
Sun Kwon Bang, Young Sik Kim, Byung Soo Chang, Cheol Beom Park, In Seok Bang
Biotechnology and Bioprocess Engineering.2013; 18(5): 835. CrossRef
- Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development
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Yan Cheng , Youjun Feng , Ping Luo , Jiang Gu , Shu Yu , Wei-jun Zhang , Yan-qing Liu , Qing-xu Wang , Quan-ming Zou , Xu-hu Mao
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J. Microbiol. 2009;47(4):498-505. Published online September 9, 2009
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DOI: https://doi.org/10.1007/s12275-009-0116-8
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162
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Shiga toxin 2 (Stx2) is a major virulence factor for enterohemorrhagic Escherichia coli (EHEC), which is encoded by λ lysogenic phage integrated into EHEC chromosome. Stx2A1, A1 subunit of Stx2 toxin has gathered extensive concerns due to its potential of being developed into a vaccine candidate. However, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. Here we report use of the prokaryotic system pET-28a::espA-Stx2A1/BL21 to carry out the fusion expression of Stx2A1 which is linked to E. coli secreted protein A (EspA) at its N-terminus. Under the IPTG induction, EspA- Stx2A1 fusion protein in the form of inclusion body was obtained successfully, whose expression level can reach about 40% of total bacterial protein at 25°C, much higher than that at 37°C. Western blot test suggested the refolded fusion protein is of excellent immuno-reactivity with both monoclonal antibodies, which are specific to EspA and Stx2A1, respectively. Anti-sera from Balb/c mice immunized with the EspA-Stx2A1 fusion protein were found to exhibit strong neutralization activity and protection capability in vitro and in vivo. These data have provided a novel feasible method to produce Stx2A1 in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against EHEC O157:H7 infections.
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Citations
Citations to this article as recorded by

- The Diversity of Escherichia coli Pathotypes and Vaccination Strategies against This Versatile Bacterial Pathogen
Pravil Pokharel, Sabin Dhakal, Charles M. Dozois
Microorganisms.2023; 11(2): 344. CrossRef - In silico design, production and immunization evaluation of a recombinant bivalent fusion protein candidate vaccine against E. coli O157:H7
Hossein Samiei, Shahram Nazarian, Abass Hajizade, Emad Kordbacheh
International Immunopharmacology.2023; 114: 109464. CrossRef - Oral Administration with Live Attenuated Citrobacter rodentium Protects Immunocompromised Mice from Lethal Infection
Shuyu Wang, Xue Xia, Yue Liu, Fengyi Wan, Guy H. Palmer
Infection and Immunity.2022;[Epub] CrossRef - Diagnosis and Treatment for Shiga Toxin-Producing Escherichia coli Associated Hemolytic Uremic Syndrome
Yang Liu, Hatim Thaker, Chunyan Wang, Zhonggao Xu, Min Dong
Toxins.2022; 15(1): 10. CrossRef - Immunization of mice with chimeric antigens displaying selected epitopes confers protection against intestinal colonization and renal damage caused by Shiga toxin-producing Escherichia coli
David A. Montero, Felipe Del Canto, Juan C. Salazar, Sandra Céspedes, Leandro Cádiz, Mauricio Arenas-Salinas, José Reyes, Ángel Oñate, Roberto M. Vidal
npj Vaccines.2020;[Epub] CrossRef - Treatment Strategies for Infections With Shiga Toxin-Producing Escherichia coli
Sabrina Mühlen, Petra Dersch
Frontiers in Cellular and Infection Microbiology.2020;[Epub] CrossRef - Oral immunization of mice with Lactococcus lactis expressing Shiga toxin truncate confers enhanced protection against Shiga toxins of Escherichia coli O157:H7 and Shigella dysenteriae
Sagi Sreerohini, Konduru Balakrishna, Manmohan Parida
APMIS.2019; 127(10): 671. CrossRef - Mucosal and systemic responses of immunogenic vaccines candidates against enteric Escherichia coli infections in ruminants: A review
A. Lawan, F.F.A. Jesse, U.H. Idris, M.N. Odhah, M. Arsalan, N.A. Muhammad, K.R. Bhutto, I.D. Peter, G.A. Abraham, A.H. Wahid, M.L. Mohd-Azmi, M. Zamri-Saad
Microbial Pathogenesis.2018; 117: 175. CrossRef - Intestinal Pathogenic Escherichia coli: Insights for Vaccine Development
Maricarmen Rojas-Lopez, Ricardo Monterio, Mariagrazia Pizza, Mickaël Desvaux, Roberto Rosini
Frontiers in Microbiology.2018;[Epub] CrossRef - Current pathogenic Escherichia coli foodborne outbreak cases and therapy development
Shih-Chun Yang, Chih-Hung Lin, Ibrahim A. Aljuffali, Jia-You Fang
Archives of Microbiology.2017; 199(6): 811. CrossRef - In silico analysis of Shiga toxins (Stxs) to identify new potential vaccine targets for Shiga toxin-producing Escherichia coli
Maryam Golshani, Mana Oloomi, Saeid Bouzari
In Silico Pharmacology.2017;[Epub] CrossRef - Production and purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro) with high-purity and low endotoxin content
Douglas B. Figueiredo, Eneas Carvalho, Mauricio P. Santos, Stefanie Kraschowetz, Rafaela T. Zanardo, Gilson Campani, Gabriel G. Silva, Cíntia R. Sargo, Antonio Carlos L. Horta, Roberto de C. Giordano, Eliane N. Miyaji, Teresa C. Zangirolami, Joaquin Cabre
Applied Microbiology and Biotechnology.2017; 101(6): 2305. CrossRef - Escherichia coli outer membrane protein F (OmpF): an immunogenic protein induces cross-reactive antibodies against Escherichia coli and Shigella
Xiao Wang, Da Teng, Qingfeng Guan, Ruoyu Mao, Ya Hao, Xiumin Wang, Junhu Yao, Jianhua Wang
AMB Express.2017;[Epub] CrossRef -
Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for
Shigella
,
Salmonella,
enterotoxigenic
E. coli
(ETEC) enterohemorragic
Miguel O’Ryan, Roberto Vidal, Felipe del Canto, Juan Carlos Salazar, David Montero
Human Vaccines & Immunotherapeutics.2015; 11(3): 601. CrossRef - Recombinant outer membrane protein A induces a protective immune response against Escherichia coli infection in mice
Qingfeng Guan, Xiao Wang, Xiumin Wang, Da Teng, Ruoyu Mao, Yong Zhang, Jianhua Wang
Applied Microbiology and Biotechnology.2015; 99(13): 5451. CrossRef - Paving the way to construct a new vaccine against Escherichia coli from its recombinant outer membrane protein C via a murine model
Xiao Wang, Qingfeng Guan, Xiumin Wang, Da Teng, Ruoyu Mao, Junhu Yao, Jianhua Wang
Process Biochemistry.2015; 50(8): 1194. CrossRef - Comparative Genomics and Immunoinformatics Approach for the Identification of Vaccine Candidates for Enterohemorrhagic Escherichia coli O157:H7
Víctor A. García-Angulo, Anjana Kalita, Mridul Kalita, Luis Lozano, Alfredo G. Torres, S. M. Payne
Infection and Immunity.2014; 82(5): 2016. CrossRef - Advances in the development of enterohemorrhagic Escherichia coli vaccines using murine models of infection
Victor A. Garcia-Angulo, Anjana Kalita, Alfredo G. Torres
Vaccine.2013; 31(32): 3229. CrossRef - Evaluation of hha and hha sepB mutant strains of Escherichia coli O157:H7 as bacterins for reducing E. coli O157:H7 shedding in cattle
Vijay K. Sharma, Evelyn A. Dean-Nystrom, Thomas A. Casey
Vaccine.2011; 29(31): 5078. CrossRef - Vaccination of attenuated EIS-producing Salmonella induces protective immunity against enterohemorrhagic Escherichia coli in mice
Jiang Gu, Yuanyuan Ning, Haiguang Wang, Daping Xiao, Bin Tang, Ping Luo, Yan Cheng, Mingming Jiang, Na Li, Quanming Zou, Xuhu Mao
Vaccine.2011; 29(43): 7395. CrossRef
- Stable Expression and Secretion of Polyhydroxybutyrate Depolymerase of Paucimonas lemoignei in Escherichia coli
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Se Whan Park , Moon Gyu Chung , Hwa Young Lee , Jeong Yoon Kim , Young Ha Rhee
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J. Microbiol. 2008;46(6):662-669. Published online December 24, 2008
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DOI: https://doi.org/10.1007/s12275-008-0283-z
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177
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An efficient strategy for the expression and secretion of extracellular polyhydroxybutyrate depolymerase (PhaZ1) of Paucimonas lemoignei in Escherichia coli was developed by employing the signal peptide of PhaZ1 and a truncated ice nucleation protein anchoring motif (INPNC). Directly synthesized mature form of PhaZ1 was present in the cytoplasm of host cells as inclusion bodies, while a construct containing PhaZ1 and its own N-terminal signal peptide (PrePhaZ1) enabled the secretion of active PhaZ1 into the extracellular medium. However, the PrePhaZ1 construct was harmful to the host cell and resulted in atypical growth and instability of the plasmid during the cultivation. In contrast, INPNC-PhaZ1 and INPNCPrePhaZ1 fusion constructs did not affect growth of host cells. INPNC-PhaZ1 was successfully displayed on the cell surface with its fusion form, but did not retain PhaZ1 activity. In the case of INPNC-PrePhaZ1, the initially synthesized fusion form was separated by precise cleavage of the signal peptide, and active PhaZ1 was consequently released into the culture medium. The amount of PhaZ1 derived from E. coli (INPNC-PrePhaZ1) was almost twice as great as that directly expressed from E. coli (PrePhaZ1), and was predominantly (approximately 85%) located in the periplasm when cultivated at 22°C but was efficiently secreted into the extracellular medium when cultivated at 37°C.
- Expression of a Recombinant Cry1Ac Crystal Protein Fused with a Green Fluorescent Protein in Bacillus thuringiensis subsp. kurstaki Cry-B
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Jong Yul Roh , In Hee Lee , Ming Shun Li , Jin Hee Chang , Jae Young Choi , Kyung Saeng Boo , Yeon Ho Je
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J. Microbiol. 2004;42(4):340-345.
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DOI: https://doi.org/2101 [pii]
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To investigate the co-expression and crystallization of a fusion gene between the Bacillus thuringiensis crystal protein and a foreign protein in B. thuringiensis, the expression of the Cry1Ac fused with green fluorescent protein (GFP) genes in a B. thuringiensis Cry-B strain was examined. The cry1Ac gene was cloned in the B. thuringiensis-E. coli shuttle vector, pHT3101, under the control of the native cry1Ac gene promoter, while the GFP gene was inserted into the XhoI site upstream of the proteolytic cleavage site, in the middle region of the cry1Ac gene (pProAc-GFP). The B. thuringiensis Cry-B strain carrying pProAc-GFP (ProAc-GFP/CB) did not produce any inclusion bodies. However, the transformed strain expressed fusion protein forms although the expression level was relatively low. Furthermore, an immunoblot analysis using GFP and Cry1Ac antibodies showed that the fusion protein was not a single species, but rather multiple forms. In addition, the N-terminal fragment of Cry1Ac and a non-fused GFP were also found in the B. thuringiensis Cry-B strain after autolysis. The sporulated cells before autolysis and the spore-crystal mixture after autolysis of ProAc-GFP/CB exhibited insecticidal activities against Plutella xylostella larvae. Accordingly, the current results suggest that a fusion crystal protein produced by the transfomant, ProAc-GFP/CB, can be functionally expressed but easily degraded in B. thuringiensis.
- Identification of a cellular protein interacting with murine retrovirus Gag polyproteins
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Choi , Won Ja
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J. Microbiol. 1996;34(4):311-315.
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The retroviral Gag polyprotein directs the assembly of virion particles and plays an important role in some events after entry into a host cell. The Gag polyprotein of a virus mixture is responsible for inducing murine acquired immunodeficiency syndrome (MAIDS) when injected into susceptible strains of mice. In order to identify the host cellular proteins which interact with the MAIDS virus Gag proteins and possibly mediate the function of the Gag proteins, mouse T-cell leukemic cDNA expression library was screened using the yeast GAL4 two hybrid system. Of 11 individual positive clones, the clone Y1 was selected for the study of protein-protein interaction. Its DNA sequence revealed that it was an exact match to the murine SH3 domain-containing protein SH3P8. It is expressed as 2.4 kbp transcripts in testis at higher levels and in various tissues tested at lower levels. Glutathione S-transferase-Y1 fusion protein binds tightly to Pr60^def-gag as well as Pr 65^eco-gag.
- Production of the Nucleocapsid Protein of Newcastle Disease Virus in Escherichia coli and its Assembly into Ring- and Nucleocapsid-like Particles
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Chiew Ling Kho , Wen Siang Tan , Khatijah Yusoff
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J. Microbiol. 2001;39(4):293-299.
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The nucleocapsid (NP) protein of Newcastle disease virus (NDV) and its derivative (NP_cfus ) containing the myc region and six histidine residues fused to its C-terminus were expressed abundantly in Escherichia coli. The proteins were purified by sucrose gradient centrifugation. Both the NP and NP_cfus proteins self-assembled into ring-like particles with a diameter of 24 +- 2 nm around a central hole of 7 +- 1 nm. Some of these ring-like particles stacked together to form nucleocapsid-like structures which are heterogeneous in length with a diameter of 20 +- 2 nm and a central hollow of 5 +- 1 nm. Only a very small amount of the monomers in the particles was linked by inter-molecular disulfide bonds. Fusion of the C-terminal end to 29 amino acids inclusive of the myc epitope and His-tag did not impair ring assembly but inhibited the formation of the long herringbone structures. Immunogold labeling of the particles with the anti-myc antibody showed that the C-terminus of the NP_cfus protein is exposed on the surface of these ring-like particles.