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AAV2-PDE6B restores retinal structure and function in the retinal degeneration 10 mouse model of retinitis pigmentosa by promoting phototransduction and inhibiting apoptosis

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Neural Regeneration Research 2025年第8期20卷 2408-2419页

作者： Ruiqi Qiu Mingzhu Yang Xiuxiu Jin Jingyang Liu Weiping Wang Xiaoli Zhang Jinfeng Han Bo Lei henan Eye Hospital Henan Provincial People’s HospitalPeople’s Hospital of Zhengzhou UniversityZhengzhouHenan ProvinceChina Eye Institute Henan Academy of Innovations in Medical ScienceZhengzhouHenan ProvinceChina Branch of National Clinical Research Center for Ocular Disease Henan Provincial People’s HospitalZhengzhouHenan ProvinceChina Academy of Medical Science Zhengzhou UniversityZhengzhouHenan ProvinceChina

Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell ***,there is no effective treatment for retinitis pigmentosa caused by PDE6B ***-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis *** aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal *** do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and ***-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western ***2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 *** expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type *** Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction ***,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B ***,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B *** data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upr

关键词： apoptosis AAV2-PDE6B ERK1/2 gene therapy phototransduction proteomics rd10 retinitis pigmentosa

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Niacin alters the ruminal microbial composition of cattle under high-concentrate condition

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animal Nutrition 2017年第2期3卷 180-185页

作者： Dan Luo Yufei Gao Youyou Lu Mingren Qu Xiaowen Xiong Lanjiao Xu Xianghui Zhao Ke Pan Kehui Ouyang Jiangxi provincial Key Laboratory of animal Nutrition Jiangxi Agricultural University animal husbandry bureau of Jinxian County

To understand the effects of niacin on the ruminal microbial ecology of cattle under high-concentrate diet condition, Illumina MiSeq sequencing technology was used. Three cattle with rumen cannula were used in a 3 x 3 Latin-square design trial. Three diets were fed to these cattle during 3 periods for 3 days, respectively: high-forage diet(HF; forage-to-concentrate ratio = 80:20), high-concentrate diet(HC;forage-to-concentrate ratio=20:80), and HC supplemented with 800 mg/kg niacin(HCN). Ruminal pH was measured before feeding and every 2 h after initiating feeding. Ruminal fluid was sampled at the end of each period for microbial DNA extraction. Overall, our findings revealed that subacute ruminal acidosis(SARA) was induced and the a-diversity of ruminal bacterial community decreased in the cattle of HC group. Adding niacin in HC could relieve the symptoms of SARA in the cattle but the ruminal pH value and the Shannon index of ruminal bacterial community of HCN group were still lower than those of HF group. Whatever the diet was, the ruminal bacterial community of cattle was dominated by Bacteroidetes,Firmicutes and Proteobacteria. High-concentrate diet significantly increased the abundance of Prevotella,and decreased the abundance of Paraprevotella, Sporobacter, Ruminococcus and Treponema than *** with HC, HCN had a trend to decrease the percentage of Prevotella, and to increase the abundance of Succiniclasticum, Acetivibrio and Treponema. Increasing concentrate ratio could decrease ruminal pH value, and change the ruminal microbial composition. Adding niacin in HC could increase the ruminal pH value, alter the ruminal microbial composition.

关键词： Ruminal microbial ecology α-diversity Niacin High-concentrate diet condition Cattle

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De-novo characterization of the soft-shelled turtle Pelodiscus sinensis transcriptome using Illumina RNA-Seq technology

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2013年第1期14卷 58-67页

作者： Wei WANG Cai-yan LI Chu-tian GE Lei LEI You-ling GAO Guo-ying QIAN Zhejiang provincial Top Key Discipline of Modern Microbiology and Application College of Biological and Environmental Sciences Zhejiang Wanli University Ningbo 315100 China henan provincial bureau of animal husbandry Zhengzhou 450008 China

The soft-shelled turtle Pelodiscus sinensis is a high-profile turtle species because of its nutritional and medicinal value in Asian countries. However, little is known about the genes that are involved in formation of their nutritional quality traits, especially the molecular mechanisms responsible for unsaturated fatty acid and collagen biosynthesis. In the present study, the transcriptomes from six tissues from Pelodiscus sinensis were sequenced using an IIlumina paired-end sequencing platform. We obtained more than 47 million sequencing reads and 73954 unigenes with an average size of 754 bp by de-novo assembly. In total, 55.19% of the unigenes （40 814） had significant similarity with proteins in the National Center of Biotechnology Information （NCBI） non-redundant protein database and Swiss-Prot database （E-value 〈10-5）. Of these annotated unigenes, 9156 and 11 947 unigenes were assigned to 52 gene ontology categories （GO） and 25 clusters of orthologous groups （COG）, respectively. In total, 26496 （35.83%） unigenes were assigned to 242 pathways using the Kyoto Encyclopedia of Genes and Genomes pathway database （KEGG）. In addition, we found a number of highly expressed genes involved in the regulation of P. sinensis unsatu- rated fatty acid biosynthesis and collagen formation, including desaturases, growth factors, transcription factors, and extracellular matrix components. Our data represent the most comprehensive sequence resource available for the Chinese soft-shelled turtle and could provide a basis for new research on this turtle as well as the molecular genetics and functional genomics of other terrapins. To our knowledge, we report for the first time, the large-scale RNA se- quencing （RNA-Seq） of terrapin animals and would enrich the knowledge of turtles for future research.

关键词： Pelodiscus sinensis Illumina RNA-Seq Transcriptome Gene expression

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Tissue expression,association analysis between three novel SNPs of the RXRαgene and growth traits in Chinese indigenous cattle

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Chinese Science Bulletin 2013年第18期58卷 2053-2060页

作者： MA Yun GAO HanTing LIN Feng CHEN NingBo XU YongJie JIANG JinHang LI Fen LU FangRu ZHAO Man SHI KuiLin CHENG Ni LI JunYa College of Life Sciences Xinyang Normal University College of animal husbandry and Veterinary Medicine Henan Agricultural University animal husbandry bureau of Pingdingshan Hubei Key Laboratory of animal Embryo Engineering and Molecular Breeding Institute of Animal Husbandry and VeterinaryHubei Academy of Agricultural Science Institute of animal Science Chinese Academy of Agricultural Sciences

Retinoid X receptor(RXR) α is a member of the nuclear hormone receptor superfamily that mediates the biological effects on several hormones,vitamins,and regulates lipid,glucose and energy *** this study,the tissue expression profiles of the bovine RXRαgene and association analysis of single nucleotide polymorphisms(SNPs) with growth traits were carried out in 413 Chinese native *** expression profile was analysed in ten Jiaxian cattle tissues by real-time PCR,and the results showed that RXRαgene was abundantly expressed in adipose tissue and spleen,moderately expressed in heart,liver,lung,kidney,muscle and ***,three SNPs(T27919A,T28139C and G28142A) and five haplotypes were *** with TTG was dominant with frequency of 69.1%.Chi-square test showed all populations were in Hardy-Weinberg equilibrium(P>0.05) at the three sites except Jiaxian cattle at G28142A site and Qinchuan cattle at T27919A *** analysis of combined sites showed that the individuals with TTGA genotype had significantly higher heart girth than those with TAGG genotype(Panimals with AAGA genotype had higher body weight than those with TAGG genotype(P<0.05) in T27919A-G28142A *** girth,abdominal circumference and body weight of individuals with TCAG genotype were exceedingly higher than those with TTGG(P<0.01),TTGA and TCGG(P<0.05) in T28139C-G28142A *** T27919A-T28139C site,the individuals of TCTA and TCTT genotype had significantly higher heart girth and lower height at hip cross than those with TTTA(P<0.05),and the body weight of TCAA and TCTT genotype individuals was higher than those with TTTA(P<0.05).In conclusion,these results provided evidence that the polymorphisms of RXRαgene were associated with growth traits and might apply to Chinese indigenous yellow cattle breeding program as a possible candidate for marker-assisted selection(MAS).

关键词：单核苷酸多态性组织表达谱中国黄牛生长性状基因型关联分析 Hardy-Weinberg平衡分子标记辅助选择

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Dramatic Changes of Matrix Metalloproteinases-7 and Lysozyme in the Ulcerative Colitis of Mice Induced by Dextran Sulfate Sodium

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Journal of Integrative Agriculture 2014年第4期13卷 858-869页

作者： KANG Jing-jing ZHAO De-ming TENG Ke-dao JIAO Xi-lan WANG Ping-li SUN Zhe NI Pei-pei WANG Zhi-feng ZHANG Rui YANG Yu-rong LIANG Hong-de Pathology Laboratory College of Animal Husbandry and Veterinary Science Henan Agricultural University College of animal Medicine China Agricultural University

Ulcerative colitis （UC） is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to find out the role of matrix metalloproteinases-7 （MMP-7） and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium （DSS）. The UC model was evaluated both by disease activity index （DAI） and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 （IL-6） serum levels and large intestinal fluids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inflammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated firstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC （colorectal cancer）. The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.

关键词： matrix metalloproteinases-7 lysozyme interleukin-6 ulcerative colitis dextran sulfate sodium

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On growth performance, nutrient digestibility, blood T lymphocyte subsets, and cardiac antioxidant status of broilers

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animal Nutrition 2019年第1期5卷 68-73页

作者： Wenying Huo Min Li Jianping Wang Zhixiang Wang Yanqun Huang Wen Chen Feed Nutrition Engineering Laboratory of henan Province Henan Agricultural University College of animal Science and Technology Henan University of Animal Husbandry and Economy Institute of animal Nutrition Sichuan Agricultural University

Different lipid sources differ in the fatty acid profiles and differently affect growth performance as well as immune function of broilers. The influences of different dietary lipid sources on growth performance, nutrient digestibility, blood T lymphocyte population, and cardiac antioxidant status were investigated of broilers. A total of 360 one-day-old male broilers(BW = 44 ± 3 g) were randomized into 3 treatment groups, consisting of 6 replicates with 20 birds in each group. Broilers received standard diets supplemented with 5%(wt/wt) of lard(LD, as a control diet), sesame oil(SO), or flaxseed oil(FO).Broilers in both SO and FO treatment groups had lower(P < 0.05) feed conversion ratios from 22 to 42 d and during the overall phase compared to those in LD treatment group. Meanwhile, the apparent total tract nutrient digestibility of crude fat in SO and FO treatment groups was higher than that in LD treatment group. Both FO and SO treatments decreased(P < 0.05) abdominal fat percentage compared to LD treatment. Total triglycerides and total cholesterol in chicken blood were decreased(P < 0.05) by SO and FO treatments compared to LD treatment. Feeding broilers with FO and SO led to a decrease(P < 0.05) in blood CD4^+ T lymphocyte count and in CD4^+:CD8^+ ratio compared to LD *** oil and FO treatments increased cardiac glutathione peroxidase(P < 0.05) compared to LD treatment. It is concluded that addition of 5% SO and FO to the standard corn-soybean meal diet improved feed efficiency, increased the activities of cardiac glutathione peroxidase, and affected the T lymphocytes ratio of fast growing broilers.

关键词： Broiler Performance Polyunsaturated fatty acid Blood T lymphocyte subsets Cardiac antioxidant status

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Glycine represses endoplasmic reticulum stress-related apoptosis and improves intestinal barrier by activating mammalian target of rapamycin complex 1 signaling

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animal Nutrition 2022年第1期8卷 1-9页

作者： Ying Yang Xiaoxiao Fan Yun Ji Ju Li Zhaolai Dai Zhenlong Wu State Key Laboratory of animal Nutrition China Agricultural UniversityBeijing100193China henan Yinfa animal husbandry Co. XinzhengHenan451100China

Endoplasmic reticulum(ER)stress has been associated with the dysfunction of intestinal barrier in humans and *** have previously shown that oral administration of glycine to suckling-piglets improves ER stress-related intestinal mucosal barrier impairment and jejunal epithelial ***,the underlying mechanism remains *** this study,the protective effect and the mechanism of glycine on apoptosis and dysfunction in intestinal barrier induced by brefeldin A(BFA),an ER stress inducer,was explored in porcine intestinal epithelial cells(IPEC-1).The results showed that BFA treatment led to enhanced apoptosis and upregulation of proteins involved in ER stress signaling,including inositol-requiring enzyme 1a(IRE1a),activating transcription factor 6a(ATF6a),c-Jun N-terminal kinase(JNK),and C/EBP-homologous protein(CHOP).In addition,BFA induced a dysfunction in intestinal epithelial barrier,as evidenced by the increased paracellular permeability,decreased transepithelial electrical resistance(TEER),and reduced abundance of tight junction proteins(occludin,claudin-1,zonula occludens[ZO]-1,and ZO-2).These alterations triggered by BFA were significantly abolished by glycine treatment(P<0.05),indicating a protective effect of glycine on barrier function impaired by ER ***,we found that the regulatory effect of glycine on intestinal permeability,proteins implicated in ER stress and apoptosis,as well as the morphological alterations of the ER were reversed by *** summary,our results indicated that glycine alleviates ER stress-induced apoptosis and intestinal barrier dysfunction in IPEC-1 cells in a mammalian target of rapamycin complex 1(mTORC1)-dependent *** data provides in vitro evidence and a mechanism for the protective effect of glycine against the disruption of intestinal barrier integrity induced by ER stress.

关键词： Glycine ER stress Apoptosis mTORC1 Intestinal barrier

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Use of a Multiplex RT-PCR Assay for Simultaneous Detection of the North American Genotype Porcine Reproductive and Respiratory Syndrome Virus,Swine Influenza Virus and Japanese Encephalitis Virus

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Agricultural Sciences in China 2010年第7期9卷 1050-1057页

作者： CHEN Hong-ying WEI Zhan-yong ZHANG Hong-ying LüXiao-li ZHENG Lan-lan CUI Bao-an LIU Jinpeng ZHU Qian-lei WANG Zi-xin College of animal husbandry and Veterinary Henan Agricultural UniversityZhengzhou 450002P.R.China

A multiplex reverse transcriptase-polymerase chain reaction(multiplex RT-PCR) assay was developed and subsequently evaluated for its efficacy in the detection of multiple viral infections simultaneously,in *** primers for each of the 3 RNA viruses,North American genotype porcine reproductive and respiratory syndrome virus,Japanese encephalitis virus,and swine influenza virus,were used in the testing *** assay was shown to be highly sensitive because it could detect as little as 10-5 ng of each of the respective amplicons in a single sample containing a composite of all 3 *** assay was also effective in detecting one or more of the same viruses in various combinations in specimens,including lymph nodes,lungs,spleens,and tonsils,collected from clinically ill pigs and in spleen specimens collected from aborted pig *** results from the multiplex RT-PCR were confirmed by virus *** relative efficiency(compared to the efficiency of separate assays for each virus) and apparent sensitivity of the multiplex RT-PCR method show that this method has potential for application in routine molecular diagnostic procedures.

关键词： Japanese encephalitis virus multiplex RT-PCR porcine reproductive and respiratory syndrome virus swine influenza virus

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Porcine Interleukin-2 Expression in Insect Cells and Its Enhancement of Pig Immunity to Swine Influenza Virus Inactivated Vaccine

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Agricultural Sciences in China 2010年第8期9卷 1211-1220页

作者： CHEN Hong-ying ZHANG Hong-ying HUANG Yan-quan CUI Bao-an WANG Zhen-ya WANG Yan-bin LIU Jin-peng CHAO An-jun College of animal husbandry and Veterinary Henan Agricultural University Zhengzhou 450002 P.R.China

Mature porcine interleukin-2 （pIL-2） gene was amplified by PCR from the plasmid pGEM-T-pIL2 and cloned into the baculovirus pFastBacTM Dual vector of the Bac-to-Bac baculovirus expression system under the control of the PH promoter. Recombinant plL-2 （rpIL-2） expressed in Sf9 insect cells was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunofluorescence assay. Western blot analysis confirmed that the rpIL-2 protein had a molecular mass of 20 kDa, which was larger than the molecular mass of the mature protein predicted based on its peptide sequence. The rpIL-2 protein induced in vitro proliferation of ConA-stimulated porcine splenocytes and enhanced in vivo protective immune responses induced by vaccinating the pigs with inactivated oil emulsion vaccine against swine influenza virus. The results showed that the rpIL-2 expressed in Sf9 insect cells has immunoenhancement effects; the finding lays the foundation for the preparation of a specific recombinant IL-2 protein and the development of a novel immune adjuvant of vaccines against various infectious porcine pathogens to increase the immunoprotective efficacy of vaccines.

关键词： porcine interleukin-2 Sf9 insect cells expression inactivated vaccine swine influenza virus

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Crystal Structure of a CD163 Scavenger Receptor Cysteine-rich Domain Produced in Pichia Pastoris

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Chinese Journal of Structural Chemistry 2017年第9期36卷 1409-1417页

作者：马红芳乔松林黄明东江龙光李睿张改平 Key Laboratory of animal Immunology of the Ministry of Agriculture Henan Provincial Key Laboratory of Animal ImmunologyHenan Academy of Agricultural Sciences College of animal Science and Veterinary Medicine Henan Agricultural University College of Chemistry Fuzhou University

CD163 is an endocytic receptor specifically expressed in monocytes/macrophages and belongs to class B of the scavenger receptor cysteine-rich（SRCR） superfamily（SRCR-SF）. It plays critical roles in various physiological and pathological pathways. In this study, the fifth SRCR domain（SRCR5） of porcine CD163（pCD163） was expressed in Pichia pastoris system. The recombinant protein was homogeneous after purification by metal-affinity and size-exclusion chromatography. The purified protein was then assayed for its biological function. Single crystals of pCD163 SRCR5 were obtained under the condition of 25% PEG 4000, 0.1 M citric acid pH3.5 and its structure was finally determined at 1.80 A. All these results provide functional pCD163 SRCR5 in large quantity for future research, and enrich the structural details for CD163 and the SRCR-SF.

关键词： CD163, SRCR, SRCR-SF, X-ray crystallography

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