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| 1 | Generation of B Cell-Deficient Pigs by Highly Efficient CRISPR/Cas9-Mediated Gene Targeting显示文摘Generating B cell-deficient mutant is the first step to produce human antibody repertoires in large animal models. In this study, we applied the clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated(Cas) system to target the JHregion of the pig Ig M heavy chain gene which is crucial for B cell development and differentiation. Transfection of Ig M-targeting Cas9 plasmid in primary porcine fetal fibroblasts(PFFs) enabled inducing gene knock out(KO) in up to 53.3% of colonies analyzed, a quarter of which harbored biallelic modification, which was much higher than that of the traditional homologous recombination(HR). With the aid of somatic cell nuclear transfer(SCNT) technology, three piglets with the biallelic Ig M heavy chain gene mutation were produced. The piglets showed no antibody-producing B cells which indicated that the biallelic mutation of the Ig M heavy chain gene effectively knocked out the function of the Ig M and resulted in a B cell-deficient phenotype. Our study suggests that the CRISPR/Cas9 system combined with SCNT technology is an efficient genome-editing approach in pigs. | Fengjiao Chen Ying Wang Yilin Yuan Wei Zhang Zijian Ren Yong Jin Xiaorui Liu Qiang Xiong Qin Chen Manling Zhang Xiaokang Li Lihua Zhao Ze Li Zhaoqiang Wu Yanfei Zhang Feifei Hu Juan Huang Rongfeng Li Yifan Dai | 2015 | Journal of Genetics and Genomics2015，42，8： | 15 |
| 2 | Particulate matter 2.5 triggers airway inflammation and bronchial hyperresponsiveness in mice by activating the SIRT2-p65 pathway显示文摘Exposure to particulate matter 2.5(PM2.5)potentially triggers airway inflammation by activating nuclear factor-κB(NF-κB).Sirtuin 2(SIRT2)is a key modulator in inflammation.However,the function and specific mechanisms of SIRT2 in PM2.5-induced airway inflammation are largely understudied.Therefore,this work investigated the mechanisms of SIRT2 in regulating the phosphorylation and acetylation of p65 influenced by PM2.5-induced airway inflammation and bronchial hyperresponsiveness.Results revealed that PM2.5 exposure lowered the expression and activity of SIRT2 in bronchial tissues.Subsequently,SIRT2 impairment promoted the phosphorylation and acetylation of p65 and activated the NF-κB signaling pathway.The activation of p65 triggered airway inflammation,increment of mucus secretion by goblet cells,and acceleration of tracheal stenosis.Meanwhile,p65 phosphorylation and acetylation,airway inflammation,and bronchial hyperresponsiveness were deteriorated in SIRT2 knockout mice exposed to PM2.5.Triptolide(a specific p65 inhibitor)reversed p65 activation and ameliorated PM2.5-induced airway inflammation and bronchial hyperresponsiveness.Our findings provide novel insights into the molecular mechanisms underlying the toxicity of PM2.5 exposure.Triptolide inhibition of p65 phosphorylation and acetylation could be an effective therapeutic approach in averting PM2.5-induced airway inflammation and bronchial hyperresponsiveness. | Manling Liu Zhaoling Shi Yue Yin Yishi Wang Nan Mu Chen Li Heng Ma Qiong Wang | 2021 | Frontiers of Medicine2021，15，5： | 2 |
| 3 | Overexpressing dominant-negative FGFR2-IIIb impedes lung branching morphogenesis in pigs显示文摘Genetic studies with mouse models have shown that fibroblast growth factor receptor 2-IIIb(FGFR2-IIIb)plays crucial roles in lung development and differentiation. To evaluate the effect of FGFR2-IIIb in pig lung development, we employed somatic cell nuclear transfer(SCNT) technology to generate transgenic pig fetuses overexpressing the transmembrane(dn FGFR2-IIIb-Tm) and soluble(dn FGFR2-IIIb-HFc) forms of the dominant-negative human FGFR2-IIIb driven by the human surfactant protein C(SP-C) promoter,which was specifically expressed in lung epithelia. Eight dn FGFR2-IIIb-Tm transgenic and twelve dn FGFR2-IIIb-HFc transgenic pig fetuses were collected from three and two recipient sows, respectively.Repression of FGFR2-IIIb in lung epithelia resulted in smaller lobes and retardation of alveolarization in both forms of dn FGFR2-IIIb transgenic fetuses. Moreover, the dn FGFR2-IIIb-HFc transgenic ones showed more deterioration in lung development. Our results demonstrate that disruption of FGFR2-IIIb signaling in the epithelium impedes normal branching and alveolarization in pig lungs, which is less severe than the results observed in transgenic mice. The dn FGFR2-IIIb transgenic pig is a good model for the studies of blastocyst complementation as well as the mechanisms of lung development and organogenesis. | Qin Chen Bin Fang Ying Wang Chu Li Xiaoxue Li Ronggen Wang Qiang Xiong Lining Zhang Yong Jin Manling Zhang Xiaorui Liu Lin Li Lisha Mou Rongfeng Li Haiyuan Yang Yifan Dai | 2018 | Journal of Genetics and Genomics2018，45，3： | 2 |
| 4 | Termination of Transcription of LAT Increases the Amounts of ICP0 mRNA but Does Not Alter the Course of HSV-1 Infection in Latently Infected Murine Ganglia显示文摘On entering sensory ganglia,herpes simplex viruses 1(HSV-1)establishes a latent infection with the synthesis of a latency associated transcript(LAT)or initiates productive infection with expression of a set of immediate early viral proteins.The precise mechanisms how expression of a genes is suppressed during the latency are unknown.One mechanism that has been proposed is illustrated in the case of ICP0,a key immediate early viral regulatory protein.Specifically,the 2 kb LAT intron is complementary to the 30 terminal portion of ICP0 m RNA.To test the hypothesis that accumulation of LAT negatively affects the accumulation of ICP0 m RNA,we inserted a DNA fragment encoding two poly(A)sequences into LAT to early terminate LAT transcript without interrupting the complementary sequence of ICP0 transcript(named as SR1603).Comparisons of the parent(SR1601)and mutant(SR1603)HSV-1 viruses showed the following:Neurons harboring latent SR1603 virus accumulated equivalent amounts of viral DNA but higher amounts of ICP0 m RNA and lower amounts of LAT,when compared to neurons harboring the SR1601 virus.One notable difference between the two viruses is that viral RNA accumulation in explanted ganglia harboring SR1603 virus initiated significantly sooner than that in neurons harboring SR1601 virus,suggesting that ICP0 may act as an activator of viral gene expression in permissive cells.Collectively,these data suggest that increased ICP0 m RNA by suppressed LAT did not affect the establishment of latency in latently infected murine ganglia. | Haifang Jiang Jiaming Wu Xianjie Liu Ruitao Lu Manling Zhou Meiling Chen Yonghong Liu Grace Guoying Zhou Wenmin Fu | 2021 | Virologica Sinica2021，36，2： | 2 |
| 5 | Poaceae Orthologs of Rice OsSGL,DUF1645 Domain-Containing Genes,Positively Regulate Drought Tolerance,Grain Length and Weight in Rice显示文摘Grain yield is a polygenic trait that can be influenced by environmental factors and genetic compositions at all plant growth stages.Currently,the molecular mechanisms behind the coordination of the interaction between grain yield-related traits remain unknown.In this study,we characterized the function of four STRESS_tolerance and GRAIN_LENGTH(Os SGL)Poaceae ortholog genes that are transcribed into DUF1645 domain-containing proteins in relation to the grain length,grain weight,and drought stress-tolerance of rice.The transgenic plants with overexpressing or heterologous high levels of Poaceae OsSGL ortholog genes exhibited longer grain size than the wild type plants.Larger cells were seen in panicles of the four transgenic lines with paraffin sectioning and scanning electron microscopy analyses.In addition,four Poaceae OsSGL ortholog genes positively affected the drought tolerance of rice.Four transgenic plants displayed higher resistance to drought stress at the seedling and vegetative stages.RNA-sequencing and qRT-PCR results indicated that over-or heterologous-expression of four Poaceae OsSGL ortholog genes also affected the transcriptome of rice plants.These genes may play a role in auxin and cytokinin biosynthesis and their transduction pathways.Taken together,these results suggested that the four OsSGL orthologs have a conserved function in the regulation of stress-tolerance and cell growth by modulating hormonal biosynthesis and signaling. | LIU Kai LI Minjuan ZHANG Bin YIN Xuming XIA Xinjie WANG Manling CUI Yanchun | 2022 | Rice science2022，29，3： | 2 |
| 6 | Lipocalin-2-induced proliferative endoplasmic reticulum stress participates in Kawasaki disease-related pulmonary arterial abnormalities显示文摘Clinical cases have reported pulmonary arterial structural and functional abnormalities in patients with Kawasaki disease(KD);however,the underlying mechanisms are unclear.In this study,a KD rat model was established via the intraperitoneal injection of Lactobacillus casei cell wall extract(LCWE).The results showed that pulmonary arterial functional and structural abnormalities were observed in KD rats.Furthermore,proliferative endoplasmic reticulum stress(ER stress)was observed in the pulmonary arteries of KD rats.Notably,the level of lipocalin-2(Lcn 2),a trigger factor of inflammation,was remarkably elevated in the plasma and lung tissues of KD rats;increased Lcn 2 levels following LCWE stimulation may result from polymorphonuclear neutrophils(PMNs).Correspondingly,in cultured pulmonary artery smooth muscle cells(PASMCs),Lcn 2 markedly augmented the cleavage and nuclear localization of activating transcription factor-6(ATF6),upregulated the transcription of glucose regulated protein 78(GRP78)and neurite outgrowth inhibitor(NOGO),and promoted PASMCs proliferation.However,proapoptotic C/EBP homologous protein(CHOP)and caspase 12 levels were not elevated.Treatment with 4-phenyl butyric acid(4-PBA,a specific inhibitor of ER stress)inhibited PASMCs proliferation induced by Lcn 2 and attenuated pulmonary arterial abnormalities and right ventricular hypertrophy and reduced right ventricular systolic pressure in KD rats.In conclusion,Lcn 2 remarkably facilitates proliferative ER stress in PASMCs,which probably accounts for KD-related pulmonary arterial abnormalities. | Zhaoling Shi Yue Yin Chen Li Hui Ding Nan Mu Yishi Wang Shanshan Jin Heng Ma Manling Liu Jie Zhou | 2021 | Science China(Life Sciences)2021，64，6： | 1 |
| 7 | Prepara- tion and characterization of Cu2 O-TiO2 : Efficient pho- tocatalytic degradation of methylene blue 显示文摘 | Xu Yuehua Liang Dahui Liu Manle | 2008 | Mater Res Bull2008，43，12： | 1 |
| 8 | Factors in?uencing the somatic cell nuclear transfer ef?ciency in pigs显示文摘Using a data set from our laboratory, we assessed the effects of several factors on pig cloning ef?ciency. The results demonstrated that cells at high con?uence( > 90%) used as donor cell resulted in higher pregnancy rate, delivery rate and overall cloning ef?ciency(number of live offspring born per reconstructed embryo transferred to recipients) compared with the cells at 60% to79% con?uence and 80% to 89% con?uence. Cells with four, ?ve and six passages compromised the pregnancy and delivery rates compared with ?rst passage cells. The number of blastocysts transferred by somatic cell nuclear transfer(SCNT) did not signi?cantly affect the cloning ef?ciency, but transfer of blastocyst derived from in vitro culture 5 d after SCNT achieved a signi?cantly higher pregnancy rate compared with one to two cell SCNT embryos from overnight culture. The highest pregnancy rate, delivery rate and the largest litter size were obtained when Bama Miniature pig ?broblasts were used as donor cells and Landrace/Yorkshire hybrid gilts were used as recipients. Recipients treated with chemicals for estrus synchronization had higher pregnancy rates compared with untreated recipients. Our data might be helpful for improving SCNT ef?ciency in pigs. | Yong JIN Manling ZHANG Xinrong JU Shuang LIANG Qiang XIONG Lihua ZHAO Xiaowei NIE Daorong HOU Qiang LIU Junzheng WANG Chenyu WANG Xiaokang LI Lining ZHANG Xiaorui LIU Ying WANG Haiyuan YANG Yifan DAI Rongfeng LI | 2019 | Frontiers of Agricultural Science and Engineering2019，6，1： | 0 |
| 9 | Crystalline Germanium-Dipyrromethene Radicals:from a Delocalized Neutral to a Localized Cation显示文摘Both dipyrromethene complexes and radicals of heavier main group elements have been of high interest.However,cationic germanium radicals and dipyrromethene-based radicals of heavier main group elements are still escaped to be isolated.Herein,we report the isolation and full characterization of a neutral Ge(I)-masked dipyrromethene-based radical 3 and the first cationic Ge(III)-centered radical 5^(·+)as stable crystalline solids.3 behaves as a germanium(I)radical in its reaction with diphenyl disulfide to form the Ge-S bond,although X-ray crystallographic,EPR spectroscopic,computational studies revealed that the unpaired electron of 3 is mainly delocalized over the C_(9)N_(2)Ge backbone and the allylic radical character is also significant in 3.In contrast to 3,the spin density of 5^(·+)is mainly localized at the Ge center with minor contribution from the dipyrromethene ligand.Moreover,reduction of 5^(·+)with potassium graphite quantitatively regenerates 5,illustrating a reversible one-electron redox pair. | Yuyang Dai Manling Bao Wenjuan Wang Zhuofeng Xie Chunmeng Liu Yuanting Su | 2022 | Chinese Journal of Chemistry2022，40，20： | 0 |
| 10 | Novel CD123×CD33 bicistronic chimeric antigen receptor(CAR)-T therapy has potential to reduce escape from single-target CAR-T with no more hematotoxicity显示文摘Dear Editor,Antigen escape is responsible for resistance[1]or disease relapse[2]from single-target chimeric antigen receptor(CAR)-T therapy.Dual-target CAR-T therapy has the potential to overcome the escape problem.However,the efficacy and safety assessment of dual-target CAR-T therapy in treating acute myeloid leukemia(AML)need further investigation.Tandem CAR-T therapy has been widely used in research and clinic.However,clustering of two connected single-chain variable fragments(scFvs)[3]and the inappropriate conjugation distance[4]pose a risk of damaging tandem CAR-T cells’function. | Zhenzhen Wang Yang Lu Yu Liu Junli Mou Xiaoyu Liu Manling Chen Ying Wang Yingxi Xu Qing Rao Haiyan Xing Kejing Tang Zheng Tian Bing Wang Wei Qi Min Wang Shaowei Qiu Dongsheng Xiong Jianxiang Wang | 2023 | Cancer Communications2023，43，10： | 0 |
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