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Electrophoretic deposition of silk fibroin coatings with pre-defined architecture to facilitate precise control over drug delivery

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Bioactive Materials 2021年第11期6卷 4243-4254页

作者： Xian Cheng dingpei long Lili Chen John A.Jansen Sander C.G.Leeuwenburgh Fang Yang Department of Dentistry-Biomaterials Radboud Institute for Molecular Life SciencesRadboud University Medical CenterPhilips van Leydenlaan 256525EX Nijmegenthe Netherlands Jiangsu Key Laboratory of Oral Diseases Nanjing Medical UniversityNanjing210029PR China Institute for Biomedical Sciences Center for Diagnostics&TherapeuticsGeorgia State UniversityAtlantaGA30302USA Department of Stomatology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430022PR China

The therapeutic precision and clinical applicability of drug-eluting coatings can be substantially improved by facilitating tunable drug ***,the design of coatings which allows for precise control over drug release kinetics is still a major ***,a double-layered silk fibroin(SF)coating system was constructed by sequential electrophoretic deposition.A mixture of dissolved Bombyx mori SF(bmSF)molecules and pre-made bmSF nanospheres at different ratios was deposited as ***,this underlayer was covered by a top-layer comprising Antheraea pernyi SF(apSF)molecules(rich in arginylglycylaspartic acid,RGD)to improve the cellular response of the resulting double-layered ***,model drug doxycycline was either pre-mixed with dissolved bmSF molecules or pre-loaded into pre-made bmSF nanospheres at the same amount before their mixing and *** thickness and nanosphere content of the under-layer architecture were proportional to the deposition time and nanosphere concentration in precursor mixtures,*** surface topography,wettability,degradation rate and adhesion strength were comparable within the double-layered coating *** expected,RGD-rich apSF top-layer improved cell adhesion,spreading and proliferation compared with bmSF ***,the amount and duration of drug release increased linearly with increasing nanosphere concentration at fixed deposition time,whereas drug release amount increased linearly with increasing deposition *** results indicate that the dosage and kinetics of loaded drugs can be quantitatively tailored by altering nanosphere concentration and deposition time as main processing ***,this study illustrates the strong potential of pre-defining coating architecture to facilitate control over drug delivery.

关键词： Electrophoretic deposition Coating Drug delivery kinetics Silk fibroin Nanospheres

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An efficient and safe strategy for germ cell-specific automatic excision of foreign DNA in F1 hybrid transgenic silkworms

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Insect Science 2024年第1期31卷 28-46页

作者： dingpei long Rongpeng Liu Yang Huang Anyao Fu Yuli Zhang Zhanzhang Hao Qiang Li Hanfu Xu Zhonghuai Xiang Aichun Zhao State Key Laboratory of Resource Insects Institute of Sericulture and Systems BiologySouthwest UniversityChongqingChina Department of Biology Georgia State UniversityAtlantaGeorgiaUSA Guangxi Institute of Sericulture Science NanningGuangxiChina Department of Mathematics and Statistics Georgia State UniversityAtlantaGeorgiaUSA

The safety of transgenic technology is a major obstacle in the popularization and use of transgenic silkworms and their *** sericulture,only the first filial generation(F1)hybrid eggs produced by cross-breeding Japanese and Chinese original strains are usually used for the large-scale breeding of silkworms,but this may result in uncontrolled transgene dispersal during the popularization and application of the F1 hybrid transgenic *** address this issue,we developed a safe and efficient strategy using the GAL4/Upstream activating sequence(UAS)system,the FLP/flippase recognition target(FRT)system,and the gonad-specific expression gene promoters(RSHP1p and Nanosp)for the germ cell-specific automatic excision of foreign DNA in the F1 hybrid transgenic *** established 2 types of activator strains,R1p::GAL4-Gr and Nsp::GAL4-Gr,containing the testis-specific GAL4 gene expression cassettes driven by RSHP1p or Nanosp,respectively,and 1 type of effector strain,UAS::FLP-Rg,containing the UAS-linked FLP gene expression *** FLP recombinase-mediated sperm-specific complete excision of FRT-flanked target DNA in the F1 double-transgenic silkworms resulting from the hybridization of R1p::GAL4-Gr and UAS::FLP-Rg was 100%,whereas the complete excision efficiency resulting from the hybridization of Nsp::GAL4-Gr and UAS::FLP-Rg ranged from 13.73%to 80.3%.Additionally,we identified a gene,sw11114,that is expressed in both testis and ovary of Bombyx mori,and can be used to establish novel gonad-specific expression systems in transgenic *** strategy has the potential to fundamentally solve the safety issue in the production of F1 transgenic silkworm eggs and provides an important reference for the safety of transgenic technology in other insect species.

关键词： Bombyx mori FLP/FRT system GAL4/UAS system germ cell-specific excision safety of transgenic technology transgenic silkworm

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Robust reactive oxygen species modulator hitchhiking yeast microcapsules for colitis alleviation by trilogically intestinal microenvironment renovation

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Bioactive Materials 2024年第6期36卷 203-220页

作者： Jintao Li Jian Song Zhichao Deng Jian Yang Xiaoqin Wang Bowen Gao Yuanyuan Zhu Mei Yang dingpei long Xiaoqin Luo Mingxin Zhang Mingzhen Zhang Runqing Li Department of Radiology the First Affiliated HospitalSchool of Public HealthXi’an Jiaotong UniversityXi’anShaanxi710061China Institute of Cardiovascular Sciences Guangxi Academy of Medical Sciencesthe People’s Hospital of Guangxi Zhuang Autonomous RegionNanningGuangxi530021China School of Basic Medical Sciences Xi’an Jiaotong UniversityXi’anShaanxi710061China Department of Clinical Laboratory the First Affiliated Hospital of Xi’an Jiaotong UniversityXi’anShaanxi710061China Department of Thoracic Surgery the First Affiliated Hospital of Xi’an Jiaotong UniversityXi’anShaanxi710061China State Key Laboratory of Resource Insects Institute of Sericulture and Systems BiologySouthwest UniversityChongqing400715China Department of Gastroenterology The First Affiliated Hospital of Xi’an Medical UniversityXi’anShaanxi710077China

Ulcerative colitis(UC)is characterized by chronic inflammatory processes of the intestinal tract of unknown *** treatments lack understanding on how to effectively alleviate oxidative stress,relieve inflammation,as well as modulate gut microbiota for maintaining intestinal homeostasis *** this study,a novel drug delivery system based on a metal polyphenol network(MPN)was constructed via metal coordination between epigallocatechin gallate(EGCG)and Fe^(3+).Curcumin(Cur),an active polyphenolic compound,with distinguished anti-inflammatory activity was assembled and encapsulated into MPN to generate *** obtained Cur-MPN could serve as a robust reactive oxygen species modulator by efficiently scavenging superoxide radical(O_(2)•-)as well as hydroxyl radical(⋅OH).By hitchhiking yeast microcapsule(YM),Cur-MPN was then encapsulated into YM to obtain CM@*** findings demonstrated that CM@YM was able to protect Cur-MPN to withstand the harsh gastrointestinal environment and enhance the targeting and retention abilities of the inflamed *** administered orally,CM@YM could alleviate DSS-induced colitis with protective and therapeutic effects by scavenging ROS,reducing pro-inflammatory cytokines,and regulating the polarization of macrophages to M1,thus restoring barrier function and maintaining intestinal ***,CM@YM also modulated the gut microbiome to a favorable state by improving bacterial diversity and transforming the compositional structure to an anti-inflammatory phenotype as well as increasing the content of short-chain fatty acids(SCFA)(such as acetic acid,propionic acid,and butyric acid).Collectively,with excellent biocompatibility,our findings indicate that synergistically regulating intestinal microenvironment will be a promising approach for UC.

关键词： Ulcerative colitis Metal polyphenol network Reactive oxygen species Yeast microcapsules Intestinal microenvironment

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Insights into the regulatory characteristics of silkworm fibroin gene promoters using a modified Gal4/UAS system

Insights into the regulatory characteristics of silkworm fib...

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中国蚕学会第十届青年学术研讨会

作者：刘荣鹏 Wenhui Zeng Tingting Tan Tao Chen Qin Luo Dawei Qu Yiyun Tang dingpei long 徐汉福 State Key Laboratory of Silkworm Genome Biology College of BiotechnologySouthwest University

The silkworm Bombyx mori is a valuable insect that synthesizes bulk amounts of fibroin protein in its posterior silk gland（PSG） and weaves these proteins into silk *** mechanism by which the fibroin protein is efficiently synthesized and precisely regulated is an important aspect that has yet to be fully ***,we describe the regulatory characteristics of the promoters of fibroin protein-encoding genes,namely,fibroin heavy chain（fibH） and fibroin light chain（fibL）,using an optimized Gal4/UAS binary *** found that（i） UAS-linked enhanced green fluorescent protein（EGFP） was effectively activated in the PSGs of Gal4/UAS transgenic silkworms,and fluorescence was continuously detected in the PSGs after complete formation of silk glands.（ii） In the PSGs of fourth-and fifth-instar larvae,EGFP mRNA was detected in only day-3 to day-6 fifth-instar larvae,and the mRNA level of EGFP driven by fibL-Gal4（LG4） was higher than that of EGFP driven by fibH-Gal4（HG4）,while the EGFP protein could be detected at each day of both larval stages.（iii） High-level expression of Gal4 and UAS-linked EGFP caused a delay in PSG degradation in Gal4/UAS transgenic silkworms.（iv） At the early pupal stage,EGFP fluorescence was also detected in fat bodies of Gal4/UAS transgenic silkworms,indicating that the PSG-specific EGFP was transported into fat bodies during PSG degeneration;however,the underlying mechanism needs to be further *** study provides a modified Gal4/UAS system used for efficient tissue-specific expression of target genes in the PSGs of silkworms and provides new insights into the regulatory characteristics of the promoters of key fibroin protein-encoding genes.

关键词： Silkworm Gal4/UAS binary system fibroin protein-encoding gene promoter regulatory characteristics

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家蚕安全转基因技术研究现状与展望

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中国农业科学 2018年第2期51卷 363-373页

作者：龙定沛郝占章向仲怀赵爱春西南大学蚕学与系统生物学研究所/家蚕基因组生物学国家重点实验室/农业部蚕桑生物学与遗传育种重点实验室

转基因技术是实现基因功能研究和生物遗传改良的重要工具。转基因生物安全问题主要包括转基因操作技术安全与转基因产品安全这两个方面。近年来业内对转基因生物的安全性研究主要集中于转基因农作物、水生动物和家禽、家畜等大型动物在... 详细信息

转基因技术是实现基因功能研究和生物遗传改良的重要工具。转基因生物安全问题主要包括转基因操作技术安全与转基因产品安全这两个方面。近年来业内对转基因生物的安全性研究主要集中于转基因农作物、水生动物和家禽、家畜等大型动物在医药、农业和食品工业等领域的安全评估,而对于具有重要科研与经济价值的农业昆虫安全转基因技术研究的报道则较为有限,而且农业部至今未有转基因昆虫安全性评估的先例。家蚕(Bombyx mori)是重要的鳞翅目模式昆虫和农业经济昆虫,家蚕安全转基因技术的深入研究对于促进基础遗传学发展和推动蚕丝产业发展均具有重要价值和意义。自2000年第一例转基因家蚕建立以来,转基因技术已广泛应用于家蚕基因功能鉴定的基础研究和品种改良的应用研究领域,但转基因家蚕的安全性问题却成了限制转基因家蚕实用化应用的主要瓶颈。因此,开展家蚕安全转基因技术体系研究对促进转基因家蚕安全性评估和产业化具有重要意义。本文系统地概述了基于条件基因打靶的家蚕安全转基因技术体系的建立与研究现状,讨论了家蚕安全转基因技术的发展趋势和应用前景,以期为转基因动物特别是农业转基因昆虫的安全转基因技术建立和完善提供参考。

关键词：家蚕生物安全安全转基因技术位点特异性重组标记基因删除基因定点修饰

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Pre-assembled nanospheres in mucoadhesive microneedle patch for sustained release of triamcinolone in the treatment of oral submucous fibrosis

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中南大学学报（医学版） 2024年第8期49卷 1245-1260页

作者： CHENG Xian YANG Yanqing HUANG Junming GUO Qiuyun ZHU Wei long dingpei ZHOU Yueying FENG Hui WANG Jie LI Yusi ZHOU Jian LIU Yanping LIU Ousheng Xiangya Stomatological Hospital and Xiangya School of Stomatology Central South UniversityHunan Engineering Research Center for Oral Digital Intelligence and Personalized MedicineHunan 3D Printing Engineering Research Center of Oral CareWANG Songling Academician Workstation for Oral-maxilofacial and Regenerative MedicineCentral South UniversityChangsha 410078 Department of Stomatology Hainan General Hospital(Affiliated Hainan Hospital of Hainan Medical University)Haikou 570311 College of Mechanical and Vehicle Engineering National Engineering Research Centre for High Efficiency GrindingHunan UniversityChangsha 410082 State Key Laboratory of Resource Insects Southwest UniversityChongqing 400715 Salivary Gland Disease Center Affiliated Stomatological Hospital of Capital Medical UniversityBeijing Key Laboratory of Tooth Regeneration and Function ReconstructionBeijing Laboratory of Oral HealthBeijing 100050China

Objective:Drug-loaded mucoadhesive silk fibroin(SF)microneedle patch can overcome the limitations of low bioavailability and significant pain associated with traditional treatment methods,such as topical application or injection of triamcinolone for oral submucous fibrosis(OSF).However,these systems release the drug too quickly,failing to meet the clinical *** study aims to construct a mucoadhesive SF microneedle patch pre-assembled with silk fibroin nanospheres(SFN)and explore its ability to sustain the release of triamcinolone in the treatment of ***:SFN was pre-assembled via precipitation reaction and characterized by scanning electron microscope(SEM)for the *** particle size andζ-potential were measured by dynamic light scattering(DLS).Triamcinolone was loaded onto SFN using a diffusional post-loading *** effective loading of triamcinolone was confirmed using Fourier-transform infrared spectroscopy(FTIR).The concentration of unloaded triamcinolone was quantified by high-performance liquid *** encapsulation efficiency and loading capacity of SFN were then calculated to determine the optimal amount of drug *** SFN suspension was pre-mixed with SF solution to prepare the microneedle *** microneedle morphology was observed by *** mechanical tests were performed to evaluate the fracture force of microneedles at different nanosphere contents(5%,10%,and 20%),determining the optimal pre-mixing ***-vivo mouse oral mucosa permeation studies were performed to ascertain the insertion depth of the microneedles via histological *** adhesive top layer was synthesized using SF and tannic acid,with FTIR confirming its successful *** viscoelasticity was characterized by a rheometer,and differential scanning calorimetry analyzed thermal *** tests evaluated the interfacial bonding strength between the adhesive layer and microneedle base to ensure no detachm

关键词： oral disease oral mucosal disease oral submucous fibrosis mucoadhesive microneedle patch

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家蚕条件基因打靶技术的建立及应用研究进展

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蚕业科学 2018年第1期44卷 1-15页

作者：龙定沛郝占章刘丹丹代静向仲怀赵爱春家蚕基因组生物学国家重点实验室农业部蚕桑生物学与遗传育种重点实验室西南大学蚕学与系统生物学研究所

条件基因打靶技术是通过借助位点特异性重组酶和预先引入宿主基因组的重组酶特异识别位点,在宿主特定发育阶段的组织或细胞中实现目标基因时空特异性定点插入、删除、替换和倒位等突变操作的技术。目前条件基因打靶技术已被广泛应用于... 详细信息

条件基因打靶技术是通过借助位点特异性重组酶和预先引入宿主基因组的重组酶特异识别位点,在宿主特定发育阶段的组织或细胞中实现目标基因时空特异性定点插入、删除、替换和倒位等突变操作的技术。目前条件基因打靶技术已被广泛应用于拟南芥（Arabidopsis thaliana）、水稻（Oryza sativa）、小鼠（Mus musculus）和果蝇（Drosophila melanogaster）等高等真核模式生物的功能基因研究,并逐渐成为转基因动植物研究领域进行基因操作的强有力工具。近年来,国内外多个研究机构系统地开展了家蚕（Bombyx mori）条件基因打靶技术的开发研究,目前已经建立了较为完善的基于多种不同位点特异性重组系统的家蚕条件基因打靶技术平台。本文较为全面地介绍目前最为常用的位点特异性重组系统的组成和作用原理,系统概述基于位点特异性重组系统的家蚕条件基因打靶策略及其应用,并对家蚕条件基因打靶技术存在的主要问题和发展趋势进行讨论。

关键词：家蚕位点特异性重组条件基因打靶转基因基因操作

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桑树Na^+/H^+逆向转运蛋白基因(MnNHX1)的克隆与耐盐力表达

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林业科学 2015年第8期51卷 16-25页

作者：边晨凯龙定沛刘雪琴魏从进龚加红赵爱春西南大学家蚕基因组生物学国家重点实验室农业部蚕桑功能基因组与生物技术重点实验室重庆400716

【目的】研究川桑液泡膜型Na+/H+逆向转运蛋白(NHX)基因的功能,探究桑树耐盐机制,为植物抗逆基因工程筛选提供优良的候选基因。【方法】以川桑基因组数据库为基础,基于同源基因序列的保守性,以川桑叶片cDNA为模板克隆川桑液泡膜型NHX基... 详细信息

【目的】研究川桑液泡膜型Na+/H+逆向转运蛋白(NHX)基因的功能,探究桑树耐盐机制,为植物抗逆基因工程筛选提供优良的候选基因。【方法】以川桑基因组数据库为基础,基于同源基因序列的保守性,以川桑叶片cDNA为模板克隆川桑液泡膜型NHX基因;利用生物学软件和在线公共平台,分析所得基因编码的蛋白质序列及功能结构域,并构建系统进化树,分析与其他物种的亲缘关系。采用荧光定量PCR方法研究在NaCl胁迫条件下不同时间段‘湖桑32号’根、茎、叶等不同组织中桑树NHX1表达量的变化情况;通过构建超量表达载体,将其转化到拟南芥中,分析转基因拟南芥在Na Cl胁迫环境中的种子发芽数,根长、侧根生长情况和幼苗成活率,并对转基因拟南芥连续浇灌含高浓度Na Cl的营养液,研究过量表达NHX基因对拟南芥的影响。【结果】本研究得到1个液泡膜型NHX基因,命名为MnNHX1(Gen Bank登录号:KJ720637);该基因ORF长度为1 644bp,编码547个氨基酸残基,具有Na+/H+交换泵,且在其上游含有抑制剂氨氯吡嗪脒结合位点(LFFIYLLPPI)以及糖基化位点等结构域,TMHMM在线程序预测Mn NHX1具有12个明显的跨膜结构区;系统进化树分析结果显示,Mn NHX1具有较高的保守性,先与源于蔷薇科的桃聚合,与桑树形态学和基因组进化分析分类结果一致。荧光定量PCR试验表明,在无Na Cl胁迫条件下桑树NHX1在‘湖桑32号’根、茎、叶中均有表达;在Na Cl胁迫处理12 h后,根、茎中桑树NHX1的表达量显著增加,而后回落;而在胁迫处理24 h后,叶中桑树NHX1的表达量显著提高,随后回落。过量表达Mn NHX1的转基因拟南芥在Na Cl胁迫环境中,种子发芽率低于野生型,而根长和侧根生长情况以及幼苗成活率都优于野生型;连续浇灌含高浓度Na Cl营养液的转基因拟南芥生长状态更为优良。【结论】MnNHX1为优良的植物耐盐基因,在桑树中为组成型表达,并受Na Cl胁迫诱导,表现出组织特异性。过量表达Mn NHX1的拟南芥耐盐能力显著提高,生存在盐胁迫环境中,依然具有良好的生长和发育能力。

关键词：桑树 Na + /H +逆向转运蛋白非生物胁迫耐盐性基因功能

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一种利用不同斑纹品种混精杂交筛选转基因家蚕纯合子的方法

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蚕业科学 2018年第5期44卷 805-809页

作者：张雨丽赵爱春龙定沛闭立辉韦伟鲁成张桂征广西壮族自治区蚕业科学研究院南宁530007 西南大学蚕学与系统生物学研究所重庆400716

基因型的纯合是保证生物后代性状不发生分离的根本。通常采用连续累代自交同时进行相关性状筛选,或者旁侧PCR筛选等方法纯化转基因生物外源基因。连续累代自交的方法筛选耗时长,工作量大,效率低;旁侧PCR方法技术要求高、专一性强。本研... 详细信息

基因型的纯合是保证生物后代性状不发生分离的根本。通常采用连续累代自交同时进行相关性状筛选,或者旁侧PCR筛选等方法纯化转基因生物外源基因。连续累代自交的方法筛选耗时长,工作量大,效率低;旁侧PCR方法技术要求高、专一性强。本研究利用不同斑纹品种混精杂交,结合雄蛾测交,筛选出双亲均为纯合子的蛾圈,作为该品种的纯合品系。该方法简单易行,周期短、效率高,是一种筛选转基因家蚕纯合品系的优良方涛。

关键词：家蚕测交混精杂交纯合子转基因阳性率

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《单孔胸腔镜手术治疗肺癌中国专家共识》解读

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中国胸心血管外科临床杂志 2021年第2期28卷 137-145页

作者：董懂韩丁培曹羽钦吴晗陈凯项捷车嘉铭朱良纲杭钧彪李鹤成上海交通大学医学院附属瑞金医院胸外科上海200025

单孔胸腔镜手术是肺癌微创治疗的重要方式。2010年,单孔胸腔镜手术首次应用于肺癌治疗;2019年、2020年,欧洲共识和中国共识相继发表。最新发表的中国共识分别从单孔胸腔镜手术的定义、用于肺癌治疗的适应证、安全性及可行性、手术技巧... 详细信息

单孔胸腔镜手术是肺癌微创治疗的重要方式。2010年,单孔胸腔镜手术首次应用于肺癌治疗;2019年、2020年,欧洲共识和中国共识相继发表。最新发表的中国共识分别从单孔胸腔镜手术的定义、用于肺癌治疗的适应证、安全性及可行性、手术技巧、学习曲线、短期及长期结果等方面为单孔胸腔镜手术标准化提供建议,对提高手术质量及减少相关并发症的发生至关重要。中国共识也对剑突下单孔胸腔镜、不插管麻醉单孔胸腔镜的现状进行了讨论,考虑到两者的技术难度与挑战性,其在临床治疗中的应用仍有一定限制。本文旨在对中国共识进行解读并比较其与欧洲共识的异同,为广大胸外科同仁提供参考。

关键词：单孔胸腔镜手术肺癌专家共识临床解读

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