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D-dopachrome tautomerase from Japanese sea bass(Lateolabrax japonicus) is a chemokine-like cytokine and functional homolog of macrophage migration inhibitory factor

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Zoological Research 2020年第1期41卷 39-50页

作者： Feng Xu ming-yun li Jiong Chen State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products Ningbo UniversityNingboZhejiang 315211China Laboratory of Biochemistry and Molecular Biology School of Marine SciencesNingbo UniversityNingboZhejiang 315832China Key Laboratory of Applied Marine Biotechnology of Ministry of Education Ningbo UniversityNingboZhejiang 315832China

D-dopachrome tautomerase(DDT),a member of the macrophage migration inhibitory factor(MIF)protein superfamily,is a newly described cytokine with chemokine-like ***,research on fish DDT remains *** this study,we identified a DDT homolog(LjDDT)from the Japanese sea bass,Lateolabrax *** analysis showed that LjDDT had typical sequence features of known DDT and MIF homologs and was most closely related to DDT of rock bream(Oplegnathus fasciatus).LjDDT transcripts were detected in all tested tissues of healthy Japanese sea bass,with the highest expression found in the *** infection with Vibrio harveyi,LjDDT transcripts were significantly down-regulated in the three tested tissues,including the liver,spleen,and head *** LjDDT(rLjDDT)and the corresponding antibody(anti-rLjDDT)were subsequently *** administration of 100μg/g anti-rLjDDT had a statistically significant protective effect on the survival of ***-infected ***,rLjDDT was able to induce the migration of monocytes/macrophages(MO/MФ)and lymphocytes both in vitro and in vivo,but without significant influence on the migration of *** exhibited chemotactic activity for lipopolysaccharide(LPS)-stimulated M1-type MO/MΦin vitro,but not for cAMP-stimulated M2-type MO/MΦ.Furthermore,the knockdown of LjCD74,but not LjCXCR4,significantly down-regulated the rLjDDT-enhanced migration of MO/MΦand relieved the rLjMIF-inhibited migration of MO/MΦ.These results indicate that LjCD74 may be the major chemotactic receptor of LjDDT and LjMIF in Japanese sea bass MO/MΦ.Combined rLjDDT+rLjMIF treatment had no significant effect on the migration of MsiRNA,LjCD74si-,or LjCXCR4sitreated MO/MΦcompared to the control group,suggesting that the roles of LjDDT and LjMIF may be *** conclusion,our study demonstrates for the first time that DDT may play a role in the immune responses of fish against bacterial infection through chemotactic

关键词： Cell migration D-dopachrome tautomerase Japanese sea bass Macrophage migration inhibitory factor Monocyte/macrophage

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Molecular cloning, pathologically-correlated expression and functional characterization of the colony- stimulating factor 1 receptor （CSF-1R） gene from a teleost, Plecoglossus altivelis

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Zoological Research 2016年第2期37卷 96-102页

作者： Qiang CHEN Xin-Jiang LU ming-yun li Jiong CHEN Laboratory of Biochemistry and Molecular Biology School of Madne Sciences Ningbo University Ningbo 315211 China Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture Ningbo University Ningbo 315211 China

Colony-stimulating factor 1 receptor （CSF-1R） is an important regulator of monocytes/macrophages （MO/MФ）. Although several CSF-1R genes have been identified in teleosts, the precise role of CSF- 1R in ayu （Plecoglossus altivelis） remains unclear. In this study, we characterized the CSF-1R homologue from P. altivelis, and named it PaCSF-1R. Multiple sequence alignment and phylogenetic tree analysis showed that PaCSF-1R was most closely related to that of Japanese ricefish （Oryzias latipes）. Tissue distribution and expression analysis showed that the PaCSF-1R transcript was mainly expressed in the head kidney-derived MO/MФ, spleen, and head kidney, and its expression was significantly altered in various tissues upon Vibrio anguillarum infection. After PaCSF-1R neutralization for 48 h, the phagocytic activity of MO/MФ was significantly decreased, suggesting that PaCSF-1R plays a role in regulating the phagocytic function of ayu MO/M（P.

关键词： Colony-stimulating factor 1 receptor Pathologically-correlated expression Monocytes/macrophages Phagocytosis Sequence analysis

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A soluble FcγR homolog inhibits Ig M antibody production in ayu spleen cells

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Zoological Research 2019年第5期40卷 404-415页

作者： Kai Chen Yu-Hong Shi Jiong Chen ming-yun li Laboratory of Biochemistry and Molecular Biology School of Marine Sciences Ningbo University Ningbo Zhejiang 315832 China State Key Laboratory for Quality and Safety of Agro-products Ningbo University Ningbo Zhejiang 315211 China

Classical Fc receptors (FcRs) mediate the binding to and recognition of the Fc portion of antibodies and play an important role during immune responses in mammals. Although proteins similar to soluble FcRs have been identified in fish, little is known about the role of such proteins in fish immunity. Here, we cloned a cDNA sequence encoding a soluble Fc receptor for an immunoglobulin G (FcγR) homolog from ayu (Plecoglossus altivelis)(PaFcγRl). The predicted protein was composed of two immunoglobulin C2-like domains but lacked a transmembrane segment and a cytoplasmic tail. The PaFcγRl transcripts were distributed at low levels in all tested tissues, but significantly increased after Vibrio anguillarum infection. The PaFcγRl protein was expressed in the head kidney, trunk kidney, and neutrophils. Recombinant PaFcγRl (rPaFcγRl) was secreted when transfected into mammalian cells and the native protein was also detected in serum upon infection. rPaFcγRl was also demonstrated to bind to ayu IgM, as assessed by cell transfection. Suppressive activity of the recombinant mature protein of PaFcγRl (rPaFcγRlm) on in vitro anti-sheep red blood cell (SRBC) responses was detected by a modified hemolytic plaque forming cell assay. In conclusion, our study revealed that PaFcγRl is closely involved in the negative regulation of IgM production in the ayu spleen.

关键词： Soluble FcγR homolog Sequence characterization IgM-binding protein Inhibition of IgM production Plecoglossus altivelis

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Role of sortase in Streptococcus mutans under the effect of nicotine

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International Journal of Oral Science 2013年第4期5卷 206-211页

作者： ming-yun li Rui-Jie Huang Xue-Dong Zhou Richard L Gregory School of Dentistry Indiana University State Key Laboratory of Oral Diseases West China Hospital of StomatologySichuan University Department of Pathology and Laboratory Medicine School of MedicineIndiana University

Streptococcus mutans is a common Gram-positive bacterium and plays a significant role in dental caries. Tobacco and/or nicotine have documented effects on S. mutans growth and colonization. Sortase A is used by many Gram-positive bacteria, including S. mutans, to facilitate the insertion of certain cell surface proteins, containing an LPXTGX motif such as antigen 1/11. This study examined the effect of nicotine on the function of sortase A to control the physiology and growth of S. mutans using wild-type S. mutans NG8, and its isogenic sortase-defective and -complemented strains. Briefly, the strains were treated with increasing amounts of nicotine in planktonic growth, biofilm metabolism, and sucrose-induced and saliva-induced antigen I/ll-dependent biofilm formation assays. The strains exhibited no significant differences with different concentrations of nicotine in planktonic growth assays. However, they had significantly increased （P~〈 0.05） biofilm metabolic activity （2- to 3-fold increase） as the concentration of nicotine increased. Furthermore, the sortase-defective strain was more sensitive metabolically to nicotine than the wild-type or sortase-complemented strains. All strains had significantly increased sucrose-induced biofilm formation （2- to 3-fold increase） as a result of increasing concentrations of nicotine. However, the sortase-defective strain was not able to make as much sucrose- and saliva-induced biofilm as the wild-type NG8 did with increasing nicotine concentrations. These results indicated that nicotine increased metabolic activity and sucrose-induced biofilm formation. The saliva-induced biofilm formation assay and qPCR data suggested that antigen 1/11 was upregulated with nicotine but biofilm was not able to be formed as much as wild-type NG8 without functional sortase A.

关键词： dental caries metabolism nicotine sortase A Streptococcus mutans

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Mudskipper interleukin-34 modulates the functions of monocytes/macrophages via the colony-stimulating factor-1 receptor 1

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Zoological Research 2020年第2期41卷 123-137页

作者： Hai-Yu Shen Yan Zhou Qian-Jin Zhou ming-yun li Jiong Chen State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products Ningbo UniversityNingboZhejiang 315211China Laboratory of Biochemistry and Molecular Biology School of Marine SciencesNingbo UniversityNingboZhejiang 315832China Key Laboratory of Applied Marine Biotechnology of Ministry of Education Ningbo UniversityNingboZhejiang 315832China

Interleukin-34(IL-34)is a novel cytokine that plays an important role in innate immunity and inflammatory processes by binding to the colonystimulating factor-1 receptor(CSF-1R).However,information on the function of IL-34 in fish remains *** the present study,we identified an IL-34 homolog from mudskippers(Boleophthalmus pectinirostris).In silico analysis showed that the mudskipper IL-34(BpIL-34)was similar to other known IL-34 variants in sequence and structure and was most closely related to an orange-spotted grouper(Epinephelus coioides)***-34 transcripts were constitutively expressed in various tissues,with the highest level of expression found in the *** tarda infection significantly up-regulated the mRNA expression of BpIL-34 in the mudskipper *** recombinant mature BpIL-34 peptide(rBpIL-34)was purified and used to produce anti-rBpIL-34 *** blot analysis combined with PNGase F digestion revealed that native BpIL-34 in monocytes/macrophages(MOs/MФs)was *** vitro,rBpIL-34 treatment enhanced the phagocytotic and bactericidal activity of mudskipper MOs/MФs,as well as the mRNA expression of pro-inflammatory cytokines like tumor necrosis factorα(BpTNF-α)and BpIL-1βin these ***,the knockdown of mudskipper CSF-1R1(BpCSF-1R1),but not mudskipper BpCSF-1R2,significantly inhibited the rBpIL-34-mediated enhanced effect on MO/MФ*** conclusion,our results indicate that mudskipper BpIL-34 modulates the functions of MOs/MФs via BpCSF-1R1.

关键词： Interleukin-34 Mudskipper Monocyte/macrophage function Edwardsiella tarda Colonystimulating factor-1 receptor

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Comparative transcriptome analysis on the alteration of gene expression in ayu （Plecoglossus altivelis）larvae associated with salinity change

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Zoological Research 2016年第3期37卷 126-135页

作者： Xin-Jiang LU Hao ZHANG Guan-Jun YANG ming-yun li Jiong CHEN Laboratory of Biochemistry and Molecular Biology School of Marine Sciences Ningbo University Ningbo 315211 China Collaborative Innovation Center for Zhejiang Marine High-Efficiency and Healthy Aquaculture Ningbo University Ningbo 315211 China

Ayu （Plecoglossus altivelis） fish, which are an amphidromous species distributed in East Asia, live in brackish water （BW） during their larval stage and in fresh water （FW） during their adult stage. In this study, we found that FW-acclimated ayu larvae exhibited a slower growth ratio compared with that of BW-acclimated larvae. However, the mechanism underlying FW acclimation on growth suppression is poorly known. We employed transcriptome analysis to investigate the differential gene expression of FW acclimation by RNA sequencing. We identified 158 upregulated and 139 downregulated transcripts in FW-acclimated ayu larvae compared with that in BW-acclimated larvae. As determined by Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway mapping, functional annotation of the genes covered diverse biological functions and processes, and included neuroendo- crinology, osmotic regulation, energy metabolism, and the cytoskeleton. Transcriptional expression of several differentially expressed genes in response to FW acclimation was further confirmed by real-time quantitative PCR. In accordance with transcriptome analysis, iodothyronine deiodinase （ID）, pro-opiom- elanocortin （POMC）, betaine-homocysteine S-meth- yltransferase 1 （BHMT）, fructose-bisphosphate aldolase B （aldolase B）, tyrosine aminotransferase （TAT）, and Na＋-K＋ ATPase （NKA） were upregulated after FW acclimation. Furthermore, the mRNA expressions of b-type natriurefic peptide （BNP） and transgelin were downregulated after FW acclimation. Our data indicate that FW acclimation reduced the growth rate of ayu larvae, which might result from the expression alteration of genes related to endocrine hormones, energy metabolism, and direct osmoregulation.

关键词： Plecoglossus altivelis Salinity change Transcriptome analysis Growth rate Real-time quantitative PCR

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Quaternary ammonium-induced multidrug tolerant Streptococcus mutans persisters elevate cariogenic virulence in vitro

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International Journal of Oral Science 2017年第4期9卷 224-231页

作者： Ya-ling Jiang Wei Qiu Xue-Dong Zhou Hao li Jun-Zhuo Lu Hockin HK Xu Xian Peng ming-yun li ming-Ye Feng Lei Cheng Biao Ren State Key Laboratory of Oral Diseases&National Clinical Research Center for Oral Diseases&Department of Operative Dentistry and Endodontics West China Hospital of StomatologySichuan UniversityChengduChina Biomaterials&Tissue Engineering Division Department of EndodonticsProsthodontics and Operative DentistryUniversity of Maryland Dental SchoolBaltimoreUSA

Dental caries are the most prevalent chronic infections in the oral cavity, and Streptococcus mutans acts as the main cariogenic bacterial species. Antibacterial quaternary ammonium compounds （QAs） have been developed to preveFnt or treat dental caries. However, there is no report on the tolerance of S. mutans to QAs. In this study, we investigated the development of S. mutans persistence induced by a novel dental caries defensive agent, dimethylaminododecyl methacrylate （DMADDM）, for the first time. Typical biphasic killing kinetics for persisters were observed in both S. mutans planktonic and biofilm cultures challenged by DMADDM at concentrations of 20 and 200 ***- z respectively. The persisters tolerated six other antibiotics with different antibacterial mechanisms, while only daptomycin and vancomycin could slightly reduce the persister numbers in planktonic cultures. The distribution of persisters in DMADDM-treated biofilms was similar to that in the untreated control, except that the total biomass and biofilm height were significantly reduced. A higher exopolysaccharides （EPS）：bacteria ratio was observed in DMADDM-treated biofilms. Persisters in biofilms significantly upregulated gtf gene expression, indicating an increase in the bacteria＇s ability to produce EPS and an elevated capability of cariogenic virulence. Carbon source metabolism was significantly reduced, as related metabolic genes were all downregulated in persisters. Concentrations of 0.1 mM, 1 mM and 10 mM of extra glucose significantly reduced the number of persisters both in planktonic and biofilm conditions. The formation of non- inheritable and multidrug tolerant persisters induced by DMADDM suggested that drug tolerance and new persistent eradication strategies should be considered for oral antibacterial agents.

关键词： antibacterial quaternary ammoniums dental caries persistence multidrug tolerance Streptococcus mutans biofilms

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Molecular characterization of an IL-1β gene from the large yellow croaker（Larimichthys crocea） and its effect on fish defense against Vibrio alginolyticus infection

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Zoological Research 2015年第3期36卷 133-141页

作者： Jun WU Yu-Hong SHI Xue-Heng ZHANG Chang-Hong li ming-yun li Jiong CHEN Laboratory of Biochemistry and Molecular Biology School of Manne Sciences Ningbo University Ningbo 315211 China Donghai Sea Collaborative Innovation Center for Industrial Upgrading Mariculture Ningbo University Ningbo 315211 China

Interleukin 1β（IL-1β）, the first interleukin to be characterized, plays a key role in regulating the immune response. In this study, we determined the c DNA and genomic DNA sequences of the IL-1β gene from the large yellow croaker, Larimichthys crocea. Phylogenetic analysis indicated that the IL-1β（Lc IL-1β） gene was most closely related to that of European seabass（Dicentrarchus labrax）, sharing 67.8% amino acid identity. In healthy large yellow croaker, Lc IL-1β transcription was detected in all tested tissues, with the highest level found in the head kidney. Upon Vibrio alginolyticus infection, Lc IL-1β transcription in all tested tissues was significantly upregulated. Intraperitoneal injection of recombinant Lc IL-1β（r Lc IL-1β） improved the survival rate and reduced the tissue bacterial load after V. alginolyticus infection. In addition, r Lc IL-1β induced monocytes/macrophages（MO/MΦ） chemotaxis and increased phagocytosis and bactericidal activity in vitro. These results suggest that Lc IL-1β plays an important role in the large yellow croaker immune response against V. alginolyticus.

关键词： Interleukin 1β Large yellow croaker Survival rate Vibrio alginolyticus Monocytes/macrophages

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Alanine racemase is essential for the growth and interspecies competitiveness of Streptococcus mutans

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International Journal of Oral Science 2016年第4期8卷 231-238页

作者： Yuan Wei Wei Qiu Xue-Dong Zhou Xin Zheng Ke-Ke Zhang Shi-Da Wang Yu-Qing li Lei Cheng Ji-Yao li Xin Xu ming-yun li State Key Laboratory of Oral Diseases West China Hospital of Stomatology Sichuan University Department of Operative Dentistry and Endodontics West China Hospital of Stomatology Sichuan University

D-alanine （D-Ala） is an essential amino acid that has a key role in bacterial unique enzyme that interconverts L-alanine and D-alanine in most bacteria, antimicrobial drug development. Streptococcus mutans is a major causative cell wall synthesis. Alanine racemase （Air） is a making this enzyme a potential target for factor of dental caries. The factors involved in the survival, virulence and interspecies interactions of S. mutans could be exploited as potential targets for caries control. The current study aimed to investigate the physiological role of Air in S. mutans. We constructed air mutant strain of S. mutans and evaluated its phenotypic traits and interspecies competitiveness compared with the wild-type strain. We found that air deletion was lethal to S. mutans. A minimal supplement of D-Ala （150 ***- 1） was required for the optimal growth of the air mutant. The depletion of D-alanine in the growth medium resulted in cell wall perforation and cell lysis in the air mutant strain. We also determined the compromised competitiveness of the air mutant strain relative to the wild-type S. mutans against other oral streptococci （S. sanguinis or S. gordonil~, demonstrated using either conditioned medium assays or dual-species fluorescent in situ hybridization analysis. Given the importance and necessity of air to the growth and competitiveness of S. mutans, Air may represent a promising target to modulate the cariogenicity of oral biofilms and to benefit the management of dental caries.

关键词： alanine racemase biofilm D-alanine dental caries Streptococcus mutans

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Male reproductive function before and after the adjustment of the COVID-19 prevention policy:a multicenter study in China

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Asian Journal of Andrology 2024年第3期26卷 308-314页

作者： Jian-Zheng li Han Wu li Wang yun-Shan Wang Feng Kong Long-Qiao Cao ming liang Xiong Wang ming-Zhen Yuan Shandong Provincial Hospital Affiliated to Shandong First Medical University Jinan 250000China Institute of Women Children and Reproductive HealthShandong UniversityJinan 250012China Engineering Laboratory of Urinary Organ and Functional Reconstruction of Shandong Province Jinan 250000China The First People’sHospital of Jining Jining 272000China Department of Reproductive Medicine The Second Affiliated Hospital of Shandong University of Traditional Chinese MedicineJinan 250000China Reproductive Medicine Center Affiliated Yantai Yuhuangding Hospital of Qingdao UniversityYantai 264000China

At the end of 2022,the adjustment of the coronavirus disease 2019(COVID-19)pandemic control policy in China resulted in alarge-scale increase in public *** compare the fertility parameters of male patients before and after the adjustments ofthe COVID-19 pandemic control policy in China,we collected data on patients’medical histories and laboratory examinations ontheir first visits between June 2022 and March 2023 in five different *** were divided into five groups according tothe timeline of the policy *** data we collected from male patients included semen quality and serum reproductivehormone levels,and intergroup comparisons were made using the Mann–Whitney U and Chi-square *** total,16784 casesunderwent regular semen analysis,11180 had sperm morphology assessments,and 7200 had reproductive hormone *** showed declining trends in semen volume,sperm motility,and the progressive sperm motility rate after the policy *** comparison revealed an initial decrease and gradual recovery in progressive motility *** morphology analysisshowed increased neck and tail abnormalities after the policy *** significant change in hormone levels was *** the adjustment of the COVID-19 prevention policy in China,a decline in sperm motility and morphology was *** trend may gradually recover over 2 *** the policy adjustment,reproductive hormone levels were relatively stablethroughout,except for an increase in luteinizing hormone(LH).These changes in semen parameters suggest that the policyadjustment had a short-to medium-term impact on male reproductive function.

关键词： COVID-19 multicenter study reproduction SARS-CoV-2 semen analysis

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