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Chinese Journal of Natural Medicines 2018年第6期16卷 411-417页

作者： MA Li-Li YUAN Yan-Yan ZHAO Ming ZHOU Xin-Rong Tashina Jehangir WANG Fu-Yan XI Yang BU Shi-Zhong Runliang Diabetes Laboratory Diabetes Research Center School of Medicine Ningbo University

Nonalcoholic fatty liver disease(NAFLD) and type 2 Diabetes Mellitus(T2 DM) are highly prevalent diseases and are closely associated, with NAFLD being present in the majority of T2 DM patients. In Asian traditional medicine, mori Cortex is widely used for the treatment of diabetes and hyperlipidemia. However, whether it has a therapeutic effect on T2 DM associated with NAFLD is still unknown. The present study showed that the oral treatment with mori Cortex extract(MCE; 10 g·kg-1·d-1) lowered the blood lipid levels and reversed insulin resistance(IR) in high fat-diet/streptozotocin-induced type 2 diabetes in rats. The expression levels of sterol receptor element-binding protein-1 c(SREBP-1 c) and carbohydrate-responsive element binding protein(Ch REBP), which are involved in steatosis in NAFLD rats, were measured in the liver samples. MCE decreased the protein and m RNA expression levels of SREBP-1 c and Ch REBP. In conclusion, down-regulation of SREBP-1 c and Ch REBP might contribute to the protective effect of MCE on hepatic injury and IR in the rats with T2 DM associated with NAFLD.

关键词： mori Cortex extract Type 2 diabetes Non-alcoholic fatty liver Sterol receptor element-binding protein-It Carbohy-drate-responsive element binding protein

p10 genes of Bombyx mori nuclear polyhedrosis virus and Autographa californica multiple nuclear polyhedrosis virus

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Science China Chemistry 1995年第1期38卷 50-59页

作者：张耀洲吴祥甫李载平 Shanghai Institute of Biochemistry Chinese Academy of Sciences Shanghai 200031 China

EcoR I-P fragment has been cloned from Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) genomic DNA and used as a probe. 0.5-kb and 1.1-kb fragments including p10 gene from Bombyx mori nuclear polyhedrosis virus (BmNPV) have been hybridized. The p10 ORF was located in the EcoR I-R fragment. Initiation codon ATG of p 10 from BmNPV has been mutated by PCR, and the ATG region became a Bgl II site. A novel transfer vector pBmAcPV-1 has been constructed using both the p10 5’-flanking region whose initiation codon ATG has been mutated with BmNPV and the p 10 3’-flanking region of AcMNPV. The vector can recombine with not only AcMNPV DNA to express foreign gene in Sf cells, but also BmNPV DNA to express foreign gene in Bm cells. CAT gene was expressed at high level in Bm cells under the control of the mutated p10 promoter of BmNPV.

关键词： Bombyx mori nuclear polyhedrosis virus Autogmpha californica multiple nuclear polyhedrosis virus nudeotide sequence of p10 p10 transfer vector CAT gene expression.

CRISPR disruption of TCTP gene impaired normal development in the silkworm Bombyx mori

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Insect Science 2019年第6期26卷 973-982页

作者： Zu-Lian Liu Jun Xu Lin Ling Ru Zhang Peng Shang Yong-Ping Huang Faculty of Life Sciences Northwestern Polytechnical University Xi'anChina Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

The translationally controlled tumor protein (TCTP) is a highly conserved and multifunctional protein with activities ranging from cytoskeletal regulation to transcription regulation in numerous organisms. In insects, TCTP is essential for cell growth and proliferation. Recently, TCTP has been reported to affect the innate intestinal immune pathway in the Bombyx mori silkworm, a lepidopteran model insect. However, the comprehensive physiological roles of TCTP in the silkworm remain poorly understood. Here, we performed functional analysis of BmTCTP by using a binary transgenic CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/RNA-guided CRISPER-associated protein 9 nucleases) system. Disruption of BmTCTP led to developmental arrestment and subsequent lethality in third instar larvae. Histological analysis revealed that growth impairment originated from decreased cell size, and the proliferation and differentiation of intestinal epithelial cells were also affected. RNA-seq analysis revealed that genes involved in carbohydrate metabolism, lipid metabolism and digestive system pathways were significantly affected by BmTCTP depletion. Together, the results demonstrated that BmTCTP plays a key role in controlling larval growth and development.

关键词： Bombyx mori CRISPR/Cas9 TCTP

Genome-wide identification of chitin-binding proteins and characterization of BmCBP1 in the silkworm, Bombyx mori

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Insect Science 2019年第3期26卷 400-412页

作者： Zhi-Lang Li Sha Tian Huan Yang Xia Zhou Shu-Ping Xu Zi-Yu Zhang Jing Gong Yong Hou Qing-You Xia State Key Laboratory of Silkworm Genome Biology College of Biotechnology Southwest University Chongqing China

The insect cuticle plays important roles in numerous physiological functions to protect the body from invasion of pathogens, physical injury and dehydration. In this report, we conducted a comprehensive genome-wide search for genes encoding proteins with peritrophin A-type (ChtBD2) chitin-binding domain (CBD) in the silkworm, Bombyx mori. One of these genes, which encodes the cuticle protein BmCBP1, was additionally cloned, and its expression and location during the process of development and molting in B. mori were investigated. In total, 46 protein-coding genes were identified in the silkworm genome, including those encoding 15 cuticle proteins analogous to peritrophins with one CBD (CPAP1s), nine cuticle proteins analogous to peritrophins with three CBD (CPAP3s), 15 peritrophic membrane proteins (PMPs), four chitinases, and three chitin deacetylases, which contained at least one ChtBD2 domain. Microarray analysis indicated that CPAP-encoding genes were widely expressed in various tissues, whereas PMP genes were highly expressed in the midgut. Quantitative polymerase chain reaction and western blotting showed that the cuticle protein BmCBP1 was highly expressed in the epidermis and head, particularly during molting and metamorphosis. An immunofluorescence study revealed that chitin co-localized with BmCBP1 at the epidermal surface during molting. Additionally, BmCBP1 was notably up-regulated by 20-hydroxyecdysone treatment. These results provide a genome-level view of the chitin-binding protein in silkworm and suggest that BmCBP1 participates in the formation of the new cuticle during molting.

关键词： Bombyx mori chitin cuticle protein metamorphosis molting

Targeting of human aFGF gene into silkworm,Bombyx mori L. through homologous recombination

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Journal of Zhejiang University Science 2004年第6期5卷 644-650页

作者：吴小锋曹翠平 LabofGeneEngineering CollegeofAnimalSciencesZhejiangUniversityHangzhou310029China

The long-arm and short-arm genes of fibroin light chain (L-chain) of silkworm, Bombyx mori L., and the gene of human acidic fibroblast growth factor were cloned respectively and subsequently inserted into a transfer vector pVL 1392 used as a tool to target the L-chain region of the silkworm genome. Genomic DNA from their offsprings was extracted and the expected targeting was detected using polymerase chain reaction and DNA sequencing, as well as protein analysis. The results showed that positive events occurred and that the FGF gene was integrated into the L-chain locus through homologous recombination.

关键词：再结合媒介物基因目标 Bombyx mori L.蚕丝蛋白发光链 FGF基因转移基因技术

BmApontic is involved in neurodevelopment in the silkworm Bombyx mori

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Journal of Integrative Agriculture 2020年第6期19卷 1439-1446页

作者： MENG Miao YU Qi WANG Qin LIU Chun LIU Zhao-yang REN Chun-jiu CUI Wei-zheng LIU Qing-xin Laboratory of Developmental Genetics Shandong Agricultural UniversityTai'an 271018P.R.China Shandong Agricultural University Library Tai'an 271018P.R.China State Key Laboratory of Silkworm Genome Biology Southwest UniversityChongqing 400715P.R.China

The nervous system of the silkworm is vital for the development of *** achieves and maintains normal life activities by regulating the function of the organs and all kinds of physiological processes in the ***(BmApt),as an imports nt bZIP tran scripti on factor,is required for the formatio n of pigme ntation in the ***,the fun ction of BmApt in the development of the nervous system of the silkworm remains ***,we showed that amino acid seque nee of BmApt was evoluti on arily con served in its Myb/SANT motif and basic DNA bindi ng *** was expressed in the nervous system at the embry onic *** n of Bmapt by RNA interfere nee resulted in abno rmal development of ***,the expression of BmnetrinA,BmnetrinB and Bmfrazzled was decreased in the Bmapt knockdown *** results dem on strate that BmApt controls neurodevelopme nt by activati ng the expressi on of Bmnetrin and Bmfrazzled.

关键词： Bombyx mori BmAp on tic Bmnetrin Bmfrazzled n eurodevelopme nt RNA interfere nee

BmHpo mutation induces smaller body size and late stage larval lethality in the silkworm,Bombyx mori

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Insect Science 2018年第6期25卷 1006-1016页

作者： Xia Xu Hong-Lun Bi Zhong-Jie Zhang Yang Yang Kai Li Yong-Ping Huang Yong Zhang Lin He School of Life Science East China Normal UniversityShanghaiChina Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiChina

As a core member of the Hippo signaling pathway,Hpo plays a critical role in regulating growth and *** studies reported that loss of function of Hpo results in increased proliferation,reduced apoptosis and induction of tissue overgrowth in *** this study,we used CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/Casg)to study Hpo gene (BmHpo)function in the lepidopteran insect Bombyx mori,known commonly as the *** analysis of BmHpo revealed an array of deletions in *** found that BmHpo knockout resulted in defects in body size regnlation,in developmental defects and pigment accumulation and early death. Our data show that BmHpo is essential for regulation of insect growth and development and that CRISPR/Cas9technology can serve as a basis for functional analysis of target genes in lepidopteran insects.

关键词： Bombyx mori CRISPR/Cas9 developmental defects Hippo signaling path- way Hpo

Identification of a germline-expression promoter for genome editing in Bombyx mori

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Insect Science 2019年第6期26卷 991-999页

作者： Jun Xu Rong-Mei Chen Shu-Qing Chen Kai Chen Lin-Meng Tang De-Hong Yang Xu Yang Yong Zhang Hong-Sheng Song Yong-Ping Huang CAS Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant Sciences Shanghai Institute of Plant Physiology and Ecology Chinese Academy of Sciences ShanghaiChina College of Life Sciences Shanghai University ShanghaiChina Department of Biology University of NevadaRenoNevadaUSA

Identification of stage- and tissue-specific cis-regulatory elements will enable more precise genomic editing. In previous studies of the silkworm Bombyx mori, we identified and characterized several tissue- and sex-specific cisregulatory elements using transgenic technology, including a female- and fat body-specific promoter, vitellogenin, testis-specific promoters, Radial spoke head 1 (BmRl) and beta-tubulin 4 (Bmp4). Here we report a c/5-regulatory element specific for a somatic and germ cell-expressed promoter, nanos (Bmnos). We investigated activities of three truncated promoter sequences upstream of the transcriptional initiation site sequences of Bmnos in vitro (nos-0.6kb, nos-1 kb and nos-2kb) and in vivo (nos-2kb). In BmN cultured cells, all three lengths drove expression of the gene encoding enhanced green fluorescence protein (EGFP), although nos-2kb had the highest fluorescence activity. In transgenic silkworms, nos-2kb drove EGFP expression at the early embryonic stage, and fluorescence was concentrated in the gonads at later embryonic stages. In addition, this cisregulatory element was not sex differentiated. The fluorescence intensity gradually weakened following the larval developmental stage in the gonads and were broadly expressed in the whole body. The nos-2kb promoter drove the Cas9 system with efficiency comparable to that of the broad-spectrum strong IE1 promoter. These results indicate that Bmnos is an effective endogenous cisregulatory element in the early embryo and in the gonad that can be used in applications involving the clustered, regularly interspaced, short palindromic repeats (CRISPR)/Cas9 system.

关键词： Bombyx mori nanos promoter transgenic CRISPR/Cas9

Proboscipedia and Sex combs reduced are essential for embryonic labial palpus specification in Bombyx mori

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Journal of Integrative Agriculture 2020年第6期19卷 1482-1491页

作者： ZHANG Ru ZHANG Zhong-jie YU Ye HUANG Yong-ping QIAN Ai-rong TAN An-jiang Faculty of Life Sciences Northwestern Polytechnical UniversityXi'an 710072P.R.China Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032.P.R.China

Hox genes of proboscipedia(pb)and Sex combs reduced(Scr)and their cofactors are required for determi nation of in sect mouthpart identity and play important roles in in sect orga n morphoge ***,i n the lepidoptera n in sects,in cludi ng many agricultural and forest pests,the roles of these genes in determi nation of mouthpart morphogenesis are un *** we report that both BmPB(Gene ID:101740380)and BmSCR(Gene ID:692761)are essential for labial palpus specification in a lepidoptera n model in sect,the silkworm,Bombyx *** embry onic morphoge nesis,CRISPR/Cas9-mediated mutagenesis of BmPB induced transformation of labial palpus into thoracic leg and BmSCR mutation transformed labial palpus into *** of Hox-related cofactor extradenticle(BmEXD;Gene ID:692478)and its nuclear localization factor homothorax(BmHTH;Gene ID:576938425)also induced distortion of embryonic labial ***,mutagenesis of each of the four genes induced severe degeneration of spigot morphogenesis,an important part of spinneret *** real-time PCR analysis further revealed that the transcriptional interaction among these *** data thus provide no vel evidence for Hox gene regulati on of in sect embry onic patterning.

关键词： Bombyx mori Hox genes CRISPR/Cas9 embryonic patterning

Modified Cortex mori Capsules improving the successful rate of functional filtering blebs after reclinical glaucoma filtering surgery

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World Journal of Clinical Cases 2019年第21期7卷 3436-3445页

作者： Jing Yu Li-Xin Qiu Guo-Ping Qing Bo-Wen Zhao Hui Wang Beijing Tongren Eye Center Laboratory of Ophthalmology and Visual Science in Beijing Beijing Tongren HospitalCapital Medical University

BACKGROUND The major reason for filtering bleb failure or scarring of the bleb site is due to excessive scarring after glaucoma filtration surgery in the *** Chinese medicine has preeminence in the prevention of fibrosis formation through the regulation of systemic circulation and improvement of the properties of the inflammatory cells in the *** To examine the clinical efficacy of using the Modified Cortex mori Capsules(MCMC;Chinese name:Jiawei Sangbaipi Capsules)in the success rate of functional filtering blebs after glaucoma filtering surgery in clinical *** Sixty resurgery glaucoma patients were randomly divided into two groups:30 patients in surgery with the placebo group and 30 patients in surgery with the MCMC *** took either the placebo or the MCMC 2 wk before and after *** morphology and function filtering bleb,visual acuity,intraocular pressure,postoperative complications,the success rate of filtration surgery and clinical efficacy were *** Fifty patients completed the *** percentage of functional filtering blebs in the surgery plus MCMC group was 84%at 6 mo after surgery,which was higher than surgery plus placebo group(64%,P<0.05).The surgical success rate in the MCMC and placebo groups were 79%±8.3%and 57%±10.6%respectively(P<0.05).The visual acuity,intraocular pressure and the postoperative complications in the two groups had no significant *** Glaucoma filtering surgery while taking MCMC not only reduced excessive scar formation and increased the success rate of functional filtering blebs but also improved the success of glaucoma filtration operations.

关键词： Glaucoma Traditional Chinese medicine Modified Cortex mori Capsules Jiawei Sangbaipi Capsules Filtering bleb Scar formation