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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Yaofeng Li Xiangyun Chen Xiaofang Tang Chundong Zhang La Wang Peng Chen minhui Pan Cheng lu State Key Laboratory of Silkworm Genome Biology Southwest University Department of Biochemistry and Molecular Biology Chongqing Medical University Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University

Silk gland cells undergo multiple endomitotic cell cycles during silkworm larval ontogeny. Our previous study demonstrated that feeding is required for continued endomitosis in the silk gland cells of silkworm larvae. Furthermore, the insulin signaling pathway is closely related to nutritional signals. To investigate whether the insulin signaling pathway is involved in endomitosis in silk gland cells, in this study, we initially analyzed the effects of bovine insulin on DNA synthesis in endomitotic silk gland cells using 5-bromo-2’-deoxyuridine（Brd U） labeling technology, and found that bovine insulin can stimulate DNA synthesis. Insulin signal transduction is mainly mediated via phosphoinositide 3-kinase（PI3K）/Akt, the target of rapamycin（TOR） and the extracellular signal-regulated kinase（ERK） pathways in vertebrates. We ascertained that these three pathways are involved in DNA synthesis in endomitotic silk gland cells using specific inhibitors against each pathway. Moreover, we investigated whether these three pathways are involved in insulin-stimulated DNA synthesis in endomitotic silk gland cells, and found that the PI3K/Akt and TOR pathways, but not the ERK pathway, are involved in this process. These results provide an important theoretical foundation for the further investigations of the mechanism underlying efficient endomitosis in silk gland cells.

关键词： Bombyx mori silk gland DNA synthesis insulin signaling

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RNAi silencing of the SoxE gene suppresses cell proliferation in silkworm BmN4 cells

RNAi silencing of the SoxE gene suppresses cell proliferatio...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Ling Wei Zhiqing Li Daojun Cheng Takahiro Kusakabe minhui Pan Jun Duan Yonghu Wang Cheng lu State Key Laboratory of Silkworm Genome Biology Southwest University School of Life Science Southwest University Laboratory of Silkworm Science Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences

The transcription factor SoxE is mainly expressed in the gonad and involved in the regulation of gonad development and sex determination in animals. Here, we used the silkworm ovary-derived BmN4-SID1 cell line to survey the roles of the silkworm SoxE protein（BmSoxE） and predict its candidate binding targets. RNAi-mediated silencing of BmSoxE expression suppressed cell proliferation in BmN4-SID1 cells. A further cell cycle analysis revealed that this inhibition of cell proliferation was largely due to cell cycle arrest in G1 phase when BmSox E expression was blocked in BmN4-SID1 cells. Genome-wide microarray expression analyses demonstrated that the expression levels of a set of genes were significantly altered following BmSox E RNAi. More than half of these genes contained conserved binding sites for HMG box domain of the Sox proteins and were predicted to be candidate binding targets for BmSoxE. Importantly, some of the candidate targets may be associated with the effect of BmSox E on cell proliferation. Several candidate target genes showed gonad-specific expression in silkworm larvae. Taken together, these data demonstrate that BmSoxE is required for cell proliferation in silkworm BmN4-SID1 cells and provide valuable information for further investigations of the molecular control exerted by the BmSoxE protein over cell proliferation and gonad development in the silkworm.

关键词： Silkworm SoxE RNAi Cell proliferation Target

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Brainstem opioid peptidergic neurons regulate cough reflexes in mice

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The Innovation 2024年第6期5卷 50-61页

作者： Haicheng lu Guoqing Chen Miao Zhao Huating Gu Wenxuan Zheng Xiating Li Meizhu Huang Dandan Geng minhui Yu Xuyan Guan Li Zhang Huimeng Song Yaning Li Menghua Wu Fan Zhang Dapeng Li Qingfeng Wu Congping Shang Zhiyong Xie Peng Cao National Institute of Biological Sciences Beijing 102206China Tsinghua Institute of Multidisciplinary Biomedical Research Tsinghua UniversityBeijing 100084China Peking University-Tsinghua University-NIBS Joint Graduate Program School of Life SciencesTsinghua UniversityBeijing 100084China Department of Neurobiology School of Basic Medical SciencesBeijing Key Laboratory of Neural Regeneration and RepairAdvanced Innovation Center for Human Brain ProtectionCapital Medical UniversityBeijing 100069China Key Laboratory of Neural and Vascular Biology Ministry of EducationDepartment of Biochemistry and Molecular BiologyHebei Medical UniversityShijiazhuang 050011China Graduate School of Peking Union Medical College Chinese Academy of Medical SciencesBeijing 100005China State Key Laboratory of Molecular Development Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China School of Basic Medical Sciences Guangzhou National LaboratoryFifth Affiliated HospitalGuangzhou Medical UniversityGuangzhou 510799China Department of Psychological Medicine Zhongshan HospitalInstitute for Translational Brain ResearchState Key Laboratory of Medical NeurobiologyFudan UniversityShanghai 200433China

Cough is a vital defensive reflex for expelling harmful substances from the *** sensory afferents for the cough reflex have been intensively ***,the brain mechanisms underlying the cough reflex remain poorly ***,we developed a paradigm to quantitatively measure cough-like reflexes in *** this paradigm,we found that prodynorphin-expressing(Pdyn+)neurons in the nucleus of the solitary tract(NTS)are critical for capsaicin-induced cough-like *** neurons receive cough-related neural signals from Trpv1+vagal sensory *** activation of Pdyn+NTS neurons triggered respiratory responses resembling cough-like *** the divergent projections of Pdyn+NTS neurons,a glutamatergic pathway projecting to the caudal ventral respiratory group(cVRG),the canonical cough center,was necessary and sufficient for capsaicin-induced cough-like *** results reveal that Pdyn+NTS neurons,as a key neuronal population at the entry point of the vagus nerve to the brainstem,initiate cough-like reflexes in mice.

关键词： respiratory opioid cough

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CRISPR/Cas9-mediated ie-0 and ie-2 dual mutation confers resistance to BmNPV in Transgenic Silkworm

CRISPR/Cas9-mediated ie-0 and ie-2 dual mutation confers res...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

作者： Zhanqi Dong Zhigang Hu Qi Qin Feifan Dong Liang Huang Jiangqiong Long Peng Chen Cheng lu minhui Pan State Key Laboratory of Silkworm Genome Biology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture Southwest University

The CRISPR/Cas9 system has been widely used against a variety of infectious diseases as a therapeutic strategy. Here, we used the baculovirus DNA-replicating early transcriptional activators, immediate early-0(ie-0) and ie-2 genes, as double target sites to disrupt the Bombyx mori nucleopolyhedrovirus(BmNPV) genome and inhibit viral replication. We constructed a transgenic line expressing sgIE0-sgIE2 and hybridized with Cas9 transgenic lines to obtain a double positive transgenic Cas9(+)/sgIE0-sgIE2(+) line that could activate the CRISPR gene editing system. We first demonstrated that the system is capable of efficiently editing target genes and result in fragment deletions in the BmNPV genome. Mortality statistics showed that the survival rate of the transgenic Cas9(+)/sgIE0-sgIE2(+) line could reach more than 65% after inoculation with 1×10 occlusion bodies(OBs)/larva. Molecular analysis showed that BmNPV DNA replication and viral gene expression level in the transgenic Cas9(+)/sgIE0-sgIE2(+) line were significantly inhibited compared with the control Cas9(-)/sgRNA(-) line. These results indicated that IE-0 and IE-2, as baculovirus early transcriptional activators, can be used as target sites for gene therapy and that multi-gene editing may more effectively disrupt the viral genome and help breed disease resistance strain.

关键词： Bombyx mori BmNPV Transgenic CRISPR/Cas9 Antiviral therapy sgIE0-sgIE2

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Excision of Nucleopolyhedrovirus form Transgenic Silkworm using the CRISPR/Cas9 System30

Excision of Nucleopolyhedrovirus form Transgenic Silkworm us...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

作者： Zhanqi Dong Feifan Dong Xinbo Yu Liang Huang Yaming Jiang Zhigang Hu Peng Chen Cheng lu minhui Pan State Key Laboratory of Silkworm Genome Biology Southwest University College of Biotechnology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture Southwest University

CRISPR/Cas9-mediated genome engineering has been shown to efficiently suppress infection by disrupting genes of the pathogen. We recently constructed transgenic lines expressing CRISPR/Cas9 and the double sgRNA target Bombyx mori nucleopolyhedrovirus(BmNPV) immediate early-1(ie-1) gene in the silkworm, respectively, and obtained four transgenic hybrid lines by G1 generation hybridization: Cas9(-)/sgRNA(-), Cas9(+)/sgRNA(-), Cas9(-)/sgRNA(+), and Cas9(+)/sgRNA(+). We demonstrated that the Cas9(+)/sgRNA(+) transgenic lines effectively edited the target site of the BmNPV genome, and large fragment deletion was observed after BmNPV infection. Further antiviral analysis the Cas9(+)/sgRNA(+) transgenic lines shows that the median lethal dose(LD50) is 1,000-fold higher than the normal lines after inoculation with occlusion bodies(OB). The analysis of economic characters and off-target efficiency of Cas9(+)/sgRNA(+) transgenic hybrid line showed no significant difference compared with the normal lines. Our findings indicate that CRISPR/Cas9-mediated genome engineering more effectively targets the BmNPV genomes and could be utilized as an insect antiviral treatment.

关键词： Bombyx mori BmNPV CRISPR/Cas9 Transgenic Antiviral therapy

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Establishment of baculovirus-inducible CRISPR/Cas9 system for antiviral research in transgenic silkworms

Establishment of baculovirus-inducible CRISPR/Cas9 system fo...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传...

作者： Zhanqi Dong Liang Huang Feifan Dong Zhigang Hu Qi Qin Jiangqiong Long Mingya Cao Peng Chen Cheng lu minhui Pan State Key Laboratory of Silkworm Genome Biology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture Southwest University

The CRISPR/Cas9 system is a powerful genetic engineering technology that has been widely used in gene therapy, as well as in the development of novel antimicrobials and transgenic insects. However, several challenges such as lack of effective host target genes and off-target efficiency limit its application to insects. To overcome these challenges, we established a highly efficient virus-inducible CRISPR/Cas9 system in transgenic silkworms. This system includes a baculovirus-inducible promoter 39 K, which initiates transcription Cas9 protein, and U6 promoter, a target sgATAD3 A site of the ATPase family AAA domain-containing protein 3（ATAD3 A） gene. The double-positive transgenic line sgATAD3 A×39 K-Cas9（ATAD3 A-KO） was obtained by hybridization, which is inducible upon Bombyx mori nucleopolyhedrovirus（BmNPV） infection in silkworm. Further analysis showed that the BmNPV-inducible transgenic lines could significantly reduce off-target efficiency and exert no effect on economically relevant. Most importantly, this novel system can efficiently and consistently edit target genes and inhibit BmNPV replication as well as the stably expressed CRISPR/Cas9 system after inoculation with occlusion bodies（OB）. These results indicate that we have established a highly efficient BmNPV-inducible ATAD3 A-KO transgenic line with improved specificity for gene targeting and antiviral efficiency in transgenic silkworm, thereby providing insights into the treatment of infectious diseases and insect pest control.

关键词： Inducible CRISPR/Cas9 Transgenic Antiviral therapy BmNPV ATAD3A-KO

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Baculovirus LEF-11 nuclear localization signal is important for viral DNA replication

Baculovirus LEF-11 nuclear localization signal is important ...

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中国蚕学会第九届青年学术研讨会

作者： Tingting Chen Zhanqi Dong Nan Hu Zhigang Hu Feifan Dong Yaming Jiang Jun Li Peng Chen Cheng lu minhui Pan State Key Laboratory of Silkworm Genome Biology Southwest University Sichuan Provincial Suji Silkworm Farm Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of AgricultureSouthwest University

Baculovirus LEF-11 is a small nuclear protein that is involved in viral late gene transcription and DNA ***,the characteristics of its nuclear localization signal and its impact on viral DNA replication are *** the present study,systemic bioinformatics analysis showed that the baculovirus LEF-11 contains monopartite and bipartite classical nuclear localization signal sequences（cNLSs）,which were also detected in a few alphabaculovirus *** of representative LEF-11 proteins of four baculovirus genera indicated that the nuclear localization characteristics of baculovirus LEF-11 coincided with the predicted ***,Bombyx mori nucleopolyhedrovirus（BmNPV） LEF-11 could be transported into the nucleus during viral infection in the absence of a *** investigations demonstrated that the NLS of BmNPV LEF-11 is important for viral DNA *** findings of the present study indicate that the characteristics of the baculovirus LEF-11 protein and the NLS is essential to virus DNA replication and nuclear transport mechanisms.

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部分相干光与相干光在湍流中的传输特性实验研究

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强激光与粒子束 2015年第1期27卷 24-28页

作者：于诗杰龙敏慧卢芳韩香娥西安电子科技大学物理与光电工程学院西安710071

相干光经湍流传输引起光强闪烁效应,理论研究表明利用部分相干光可降低该影响,而相关的实验验证鲜有报道。利用水介质的对流湍流池模拟产生了Rytov方差为0.04~0.16的湍流,进行了弱起伏区湍流中部分相干光及相干光的传输实验,由接收光强... 详细信息

相干光经湍流传输引起光强闪烁效应,理论研究表明利用部分相干光可降低该影响,而相关的实验验证鲜有报道。利用水介质的对流湍流池模拟产生了Rytov方差为0.04~0.16的湍流,进行了弱起伏区湍流中部分相干光及相干光的传输实验,由接收光强计算出其光强闪烁指数随湍流Rytov方差的变化关系。实验结果表明：随着湍流强度的增加,部分相干光与相干光的闪烁效应均增强,但部分相干光的闪烁指数小于相干光。将实验数据与Andrews等人建立的闪烁指数理论模型进行比较,得出了较为一致的结论。

关键词：湍流池 Rytov方差部分相干光光强闪烁指数

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Baculovirus LEF-11 nuclear localization signal is important for viral DNA replication

Baculovirus LEF-11 nuclear localization signal is important ...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

Baculovirus LEF-11 is a small nuclear protein that is involved in viral late gene transcription and DNA replication. However, the characteristics of its nuclear localization signal and its impact on viral DNA replication are unknown. In the present study, systemic bioinformatics analysis showed that the baculovirus LEF-11 contains monopartite and bipartite classical nuclear localization signal sequences(cNLSs), which were also detected in a few alphabaculovirus species. Localization of representative LEF-11 proteins of four baculovirus genera indicated that the nuclear localization characteristics of baculovirus LEF-11 coincided with the predicted results. Moreover, Bombyx mori nucleopolyhedrovirus(BmNPV) LEF-11 could be transported into the nucleus during viral infection in the absence of a c NLSs. Further investigations demonstrated that the NLS of BmNPV LEF-11 is important for viral DNA replication. The findings of the present study indicate that the characteristics of the baculovirus LEF-11 protein and the NLS is essential to virus DNA replication and nuclear transport mechanisms.

关键词： Bombyx mori Baculovirus BmNPV LEF-11 Nuclear localization signal

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疝修补术前扩容治疗后应用抗生素对患者的影响

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中国普通外科杂志 2011年第3期20卷 317-320页

作者：戴旻晖陆璨中南大学湘雅医院手术室湖南长沙410008

目的探讨疝修补术手术前行扩容治疗后应用抗生素对患者的影响。方法将80例腰硬联合麻醉下疝修补患者随机分为A,B组,每组40例,两组患者均于麻醉后10 min使用抗生素,A组于扩容性补液补足生理病理需要量后使用抗生素,B组于给予生理病理需... 详细信息

目的探讨疝修补术手术前行扩容治疗后应用抗生素对患者的影响。方法将80例腰硬联合麻醉下疝修补患者随机分为A,B组,每组40例,两组患者均于麻醉后10 min使用抗生素,A组于扩容性补液补足生理病理需要量后使用抗生素,B组于给予生理病理需要量的1/2后使用抗生素,对两组患者术中自主感觉、生命体征的变化及应用升压药的情况进行比较分析。结果 A组术中自主感觉、生命体征的变化情况优于B组;在应用升压药方面,少于B组;3项临床观察指标均具有统计学意义(P<0.05)。两组切口均为甲级愈合。结论行疝修补术患者术前抗生素在补足生理病理需要量麻醉后10 min使用效果为好,可降低患者围术期的不良反应、提高患者舒适度及对手术的满意度,降低医疗费用。

关键词：疝/外科学抗生素预防麻醉,硬膜外,联合扩容性补液

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