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Four kinds of ENU-induced white spot mice and chromosome locations of the mutant genes

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Chinese Science Bulletin 2003年第24期48卷 2658-2664页

作者： WU Baojin1,2, MAO Huihua1, SHAO Yixiang1, XUE Zhengfeng1 & LI Houda1 1. comparative medicine center of yangzhou university, yangzhou 225009, China 2. Medical College of yangzhou university, yangzhou 225001, China Correspondence should be addressed to Li Houda (e-mail: houdali @***) comparative medicine center of yangzhou university Yangzhou China Medical College of yangzhou university Yangzhou China

Phenotype-driven is the name for an ap-proach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and repro-duced a total offspring of 3860. Of these descendants, 210 exhibited mutation phenotypes by screening, and more than 10 of them are hereditable. Four kinds of mutant mice, named Wbct, W-1Bao, W-2Bao, and W-3Bao, showed dominant hereditary white spot mutation with partial albinism on their belly, distal limbs and tail terminal. To map these mutant genes, 39 microsatellites, equally distributed on the mouse genome and with difference between B6 and DBA/2J (D2), were selected to scan the genome after discrimination of the white spots in the F2 mice [(B6×D2)×D2]. It is found that, the log odds score (LODS) between W-1Bao and D5Mit168 is 0.56, and the LODS of W-1Bao and D5Mit352 is 4.47. With the gradual application of microsatellites D5Mit290, D5Mit312, D5Mit308 and D5Mit356 that are close to the mutant gene, and the number of F2 mice going up to 537, the mutant W-1Bao is located between D5Mit356 and D5Mit308 on chro-mosome 5, about 42.19 cM from the centromere. In the same way, W-2Bao and W-3Bao are mapped nearby W-1Bao, and Wbct is located on chromosome 1, about 41.6 cM from the centro-mere. After searching for the mouse genome database (MGD) and performing a one-by-one study of all genes located on chromosome subregion, it is believed that the kit gene is an excellent candidate for the white spot mutations of W-1Bao, W-2Bao and W-3Bao.

关键词：染色体突变基因基因工程 ENU

Inactivated Sendai Virus Induces Apoptosis in Murine Melanoma Cells by IGF-1R Down-regulation

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Biomedical and Environmental Sciences 2013年第12期26卷 998-1002页

作者： GAO Hui XU Xiao Shuang CHEN Ze Dong ZHANG Quan XU Xiang Ming College of Clinical medicine of yangzhou university comparative medicine center Yangzhou University Suzhou polytechnic institute of agriculture

The mortality of cancer patients has considerably improved due to progress in surgery, chemotherapy and radiotherapy. However, some types of cancers, such as melanoma, remain refractory to conventional strategies. Although melanoma accounts for only 4% of all dermatological malignancies, it is responsible for 80% of mortalities from skin tumors[11. The reported survival rate of melanoma over 5 years is not yet encouraging due to its chemo-resistance and rapid metastasis. Therefore, it is necessary to develop new drugs with potent activity and weak side-effect against melanoma.

关键词： IGF Inactivated Sendai Virus Induces Apoptosis in Murine Melanoma Cells by IGF-1R Down-regulation

Inactivated Sendai Virus Induces ROS-dependent Apoptosis and Autophagy in Human Prostate Cancer Cells

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Biomedical and Environmental Sciences 2018年第4期31卷 280-289页

作者： QIAN Miao TAN Hai Ming YU Ning WANG Tao ZHANG Quan Institute of comparative medicine College of Veterinary Medicine Yangzhou University Jiangsu Co-innovation center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses Yangzhou University

Objective The current study aims to investigate the effect of Hemagglutinating virus of Japan envelope（HVJ-E） on induction of apoptosis and autophagy in human prostate cancer PC3 cells, and the underlying mechanisms. Methods PC3 cells were treated with HVJ-E at various multiplicity of infection（MOI）, and the generated reactive oxygen species（ROS）, cell viability, apoptosis, and autophagy were detected, respectively. Next, the role of ROS played in the regulation of HVJ-E-induced apoptosis and autuphagy in PC3 cells were analysed. In the end, the relationship between HVJ-E-induced apoptosis and autuophagy was investigated by using rapamycin and chloroquine. Results Flow cytometry assay revealed that HVJ-E treatment induced dose-dependent apoptosis and that the JNK and p38 MAPK signaling pathways were involved in HVJ-E-induced apoptosis in PC3 cells. In addition, HVJ-E was able to induce autophagy in PC3 cells via the class III PI3 K/beclin-1 pathway. The data also implyed that HVJ-E-triggered autophagy and apoptosis were ROS dependent. When ROS was blocked with N-acetylcysteine（NAC）, HVJ-E-induced LC3-II conversion and apoptosis were reversed. Interestingly, HVJ-E-induced apoptosis was significantly increased by an inducer of autophagy, rapamycin pretreatment, both in vitro and in vivo. Conclusion HVJ-E exerts anticancer effects via autophagic cell death in prostate cancer cells.

关键词： Inactivated Sendai virus(HVJ-E) Reactive oxygen species(ROS) Apoptosis Autophagy

A Dietary Source of High Level of Fluoroquinolone Tolerance in mcr-Carrying Gram-Negative Bacteria

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Research 2024年第3期2023卷 105-121页

作者： Tianqi Xu Dan Fang Fulei Li Zhiqiang Wang Yuan Liu Jiangsu Co-innovation center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary MedicineYangzhou UniversityYangzhou 225009China. Joint International Research Laboratory of Agriculture and Agri-Product Safety the Ministry of Education of ChinaYangzhou UniversityYangzhou 225009China. Institute of comparative medicine Yangzhou UniversityYangzhou 225009China.

The emergence of antibiotic tolerance,characterized by the prolonged survival of bacteria following antibiotic exposure,in natural bacterial populations,especially in pathogens carrying antibiotic resistance genes,has been an increasing threat to public ***,the major causes contributing to the formation of antibiotic tolerance and underlying molecular mechanisms are yet poorly ***,we show that potassium sorbate(PS),a widely used food additive,triggers a high level of fluoroquinolone tolerance in bacteria carrying mobile colistin resistance gene *** studies demonstrate that PS treatment results in the accumulation of intracellular fumarate,which activates bacterial two-component system and decreases the expression level of outer membrane protein OmpF,thereby reducing the uptake of *** addition,the supplementation of PS inhibits aerobic respiration,reduces reactive oxygen species production and alleviates DNA damage caused by bactericidal ***,we demonstrate that succinate,an intermediate product of the tricarboxylic acid cycle,overcomes PS-mediated ciprofloxacin *** multiple animal models,ciprofloxacin treatment displays failure outcomes in PS preadministrated animals,including comparable survival and bacterial loads with the vehicle *** together,our works offer novel mechanistic insights into the development of antibiotic tolerance and uncover potential risks associated with PS use.

关键词： overcome thereby additive

Homozygous lethality and heterozygous spotting due to a novel missense mutation in the mouse Kit gene

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Current Zoology 2009年第6期55卷 430-434页

作者： Baojin WU Lijing YIN Xiaoshu YIN Weiwei YANG Bing CHEN Zhengfeng XUE Peilin WU Laboratory of Experimental Animal Science Hangzhou Normal University Hangzhou 310036 China center for comparative medicine Yangzhou University Yangzhou Jiangsu Province 225009 China

N-ethyl-N-nitrosourea （ENU） mutagenesis in mice can be used to study gene function in vivo and to establish genetic mouse models of human disease. In this study, a white spotted mouse （named Kit^W-1 Bao） was obtained by ENU-induced mutagenesis. Inheritance testing showed a single-gene dominant mutation and lethality in the Kit^W-1 Bao homozygous mice. The mutation was mapped to Chromosome 5 between markers DSMit356 and DSMit308. The region contains the Kit gene, whose mutations are known to lead to pigmentation defects in mice. Sequence analysis of the Kit cDNA from Kit^W-1 Bao heterozygotes revealed an A to T missense mutation resulting in an amino acid substitution of Asp （D） by Val （V） at amino acid position 849 within a highly conserved tyrosine kinase domain. The combined phenotype displayed by the Kit^W-1 Bao heterozygous and homozygous mutant mice demonstrates the critical function of the highly conserved aspartie acid residue at position 849 in the Kit gene product

关键词： White spotting Death Mapping Identification Kit

Immobilized MHC class I chain-related protein A synergizes with IL-15 and soluble 4-1BB ligand to expand NK cells with high cytotoxicity ex vivo

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Cellular & Molecular Immunology 2010年第6期7卷 477-484页

作者： Weijuan Gong Weiming Xiao Li Qian Chunxiang Gong Maozhi Hu Xianyuan Pan Mingchun Ji Department of Immunology School of MedicineYangzhou UniversityYangzhouPR China Department of Gastroenterology The Second Affiliated HospitalYangzhou UniversityYangzhouChina Testing center Yangzhou UniversityYangzhouChina

Major histocompatibility complex(MHC)class I chain-related protein A(MICA),which is a ligand for human NKG2D,is expressed by a variety of epithelial tumor cells and promotes the activation of natural killer(NK),CD81 and cd-T *** ectopic expression of MICA on tumor cells elicits anti-tumor responses,soluble MICA downregulates the activities of *** this study,we showed that recombinant,immobilized MICA(iMICA)molecules coated on plastic wells weakly promote peripheral NK cell activation,secretion of interferon(IFN)-c and degranulation without inducing *** addition,iMICA synergized with IL-15 and soluble 4-1BB ligand(s4-1BBL)to expand NK cells 25-to 42-fold in a 13-day culture,whereas NK cells stimulated only with IL-15 and s4-1BBL expanded 10-to *** contrast toNKcells expanded by IL-15 and s4-1BBL stimulation,NKcells expanded long term in the presence of iMICA exhibited increased cytotoxicity against leukemia *** results suggest that large numbers ofNKcells with high cytotoxicity can be generated by stimulation with IL-15 and s4-1BBL in the presence of iMICA and that these cells can be used for adoptive cancer immunotherapy.

关键词： expansion IL-15 MICA natural killer 4-1BBL

Abnormal gonad development in Kit^(W-2Bao) mice caused by a Kit gene missense mutation

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Chinese Science Bulletin 2010年第36期55卷 4143-4149页

作者： WU BaoJin YIN LiJing LU ZhengLan YIN YuShu YANG WeiWei YANG Rong KANG XiaoDong LIU GuiJie YIN HongPing YU LiPing Gu MeiEr WU PeiLin Laboratory of Experimental Animal Science Hangzhou Normal University Hangzhou 310036 China comparative medicine Centre of yangzhou university Yangzhou 225000 China

KitW-2Bao mice are single-gene autosomal dominant mutation mice with a B6 background that were bred in our laboratory. Heterozygotes had morphological characteristics including albinism of the abdomen, extremities, and tail, whereas the homozygotes had albinism of the body, black eyes, and infertility. The homozygous mutants showed small, structurally abnormal gonads, and lacked germ cells. Heterozygous male mice lacked germ cells in some contorted seminiferous tubules. This mutation has been mapped at 43.8 cM from the centromere in chromosome 5 by linkage analysis and Kit has been identified as the candidate gene. After Kit full-length mRNA amplification, it was found that a G to T conversion at position 1228 in the ORF changed the 410th amino acid from V to F. This amino acid change could affect the protein’s secondary structure. Heterozygous mutant mice were intercrossed and homozygous mutant mice were bred and genotyped. We found that no primordial germ cells (PGCs) appeared in the urogenital ridge area at fetus day 11.5 in the homozygotes. The number of PGCs also significantly decreased in heterozygotes. At fetus day 15.5, the differentiation of the testis tubule structure was unclear; as well, they contained no spermatogonia. Female homozygotes contained no primordial follicles in the ovary. The numbers of PGCs and primordial follicles were significantly decreased in heterozygous mice. W-2Bao is the only mutated site in the extracellular 4th Ig-like domain and this mutant mouse model provides new material for the study of the mechanism of reproductive system development.

关键词：突变小鼠结构异常单基因套件性腺原始生殖细胞工具包杂合子缺乏

Mapping of genetic modifiers of Plcd1 in scant hair mice(snthr^(1Bao))

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Chinese Science Bulletin 2010年第35期55卷 4026-4031页

作者： WU BaoJin ZENG YongMei MAO HuiHua YIN LiJing ZHU Jie YANG WeiWei YIN XiaoShu WU PeiLin ZHANG WeiDong Laboratory of Experimental Animal Science Hangzhou Normal University Hangzhou 310036 China comparative medicine center Yangzhou University Yangzhou 225002 China The Jackson Laboratory Bar Harbor ME 04609 USA

The scant hair mutant mouse(locus symbol:snthr1Bao) is a recessive mutation that originated in an ethylnitrosourea chemical carcinogenesis study using the DBA/2J inbred *** gene responsible for the mutation was previously determined to be phospholipase C,delta 1(Plcd1;mutant allele symbol Plcd1snthr1Bao).To map the modifiers of Plcd1,an intercross(DBA/2J-snthr1Bao/snthr1Bao × C57BL/6J+/+) was *** F2 mutant progeny exhibited a variety of alopecia phenotypes;all F2 mutants(n=507) were classified into 3 groups(mild,moderate,and severe alopecia) and genotyped based on 96 microsatellites.A major QTL was identified on mouse chromosome(mChr) 15 at 12 cM with an LOD score greater than 7(P < 0.0001).Three minor QTLs were detected on mChr 2,5,and 7 at 40,84 and 48 cM,*** QTLs on mChr 7 and 15 were associated with minor alopecia while the QTLs on mChr 2 and 5 were associated with moderate to severe *** antagonistic or synergistic effects among or between the 4 QTLs were *** the functions of the 4 potential regulatory QTLs and mutant Plcd1snthr1Bao,we found that these QTLs might contribute to variations of scant hair severity by altering the Ca2+ signal pathways in mouse skin.

关键词：突变小鼠遗传修饰映射 QTL检测基因符号信号转导通路基因分型未成年人

Characterization of three African swine fever viruses from diferent clinical settings revealed a potential attenuation mechanism

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Animal Diseases 2024年第4期4卷 263-274页

作者： Jiajia Zhang Yuening Wang Kaili Zhang Dafu Deng Kai Peng Wanglong Zheng Nanhua Chen Penggang Liu Shaobin Shang Ruichao Li Jianzhong Zhu College Veterinary medicine Yangzhou UniversityYangzhouChina Joint International Research Laboratory of Agriculture and Agri‑Product Safety Yangzhou UniversityYangzhouChina comparative medicine Research Institute Yangzhou UniversityYangzhouChina Jiangsu Coinnovation center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses Yangzhou UniversityYangzhou 225009China

African swine fever(ASF)is an acute and fatal hemorrhagic disease in domestic pigs and wild boars caused by African swine fever virus(ASFV)that currently threatens the pig industry *** the 2018 ASF outbreak in China,ASFV has evolved and caused diverse clinical manifestations,such as chronic and asymptomatic ***,it is important to understand the molecular mechanisms underlying ASFV attenuation in the ***,we isolated three ASFVs from one diseased and two asymptomatic pigs by using primary porcine alveolar macrophages(PAMs)from both domestic pigs and Bama *** three ASFVs exhibited similar phenotypes in cell culture,includ‑ing cytopathic efects(CPEs),hemadsorptions(HADs),viral protein expressions and growth *** sequenc‑ing revealed that all three ASFVs were genotype II *** comparisons suggested that the disruption of the viral genes MGF360 and MGF110,rather than EP402R and EP153R,is likely involved in the potential attenuation of ASFV via the upregulation of innate immune responses.

关键词： African swine fever virus Virus isolation Genome sequence Genotype II Attenuation