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Cytopathological evaluations combined RNA and protein analyses on defined cell regions using single frozen tissue block

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Cell Research 2002年第2期12卷 117-121页

作者： HONG LI, XIAO YAN CHEN, QING You KONG, JIA LIULaboratory of Cell Biology and Molecular genetics,College of Basic Medical Sciences, Dalian Medical University, Dalian 116027, China Laboratory of Cell Biology and Molecular genetics College of Basic Medical Sciences Dalian Medical University Dalian China

The co-existence of multiple cell components in tissue samples is the main obstacle for precise molecular evaluation on defined cell types. Based on morphological examination, we developed an efficient approach for paralleled RNA and protein isolations from an identical histological region in frozen tissue section. The RNA and protein samples prepared were sufficient for RT-PCR and Western blot analyses, and the results obtained were well coincident each other as well as with the corresponding parameters revealed from immunohistochemical examinations. By this way, the sampling problem caused by cell-cross contamination can be largely avoided, committing the experimental data more specific to a denned cell type. These novel methods thus allow us to use single tissue block for a comprehensive study by integration of conventional cytological evaluations with nucleic acid and protein analyses.

关键词： RNA and protein preparation defined histological region serial frozen section gene expression.

A Profile of Native Integration Sites Used by φC31 Integrase in the Bovine Genome

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Journal of genetics and Genomics 2012年第5期39卷 217-224页

作者： Lijuan Qu Qingwen Ma Zaiwei Zhou Haiyan Ma Ying Huang Shuzhen Huang Fanyi Zeng Yitao Zeng Shanghai Institute of Medical genetics Children＇s Hospital of Shanghai Shanghai Jiao Tong University School of Medicine 24/1400 West Beijing Road Shanghai 200040 China Key Lab of Medical Embryo Molecular Biology Ministry of Health Shanghai 200040 China Shanghai Key Lab of Embryo and Reproduction Engineering Shanghai Municipal Committee of Science and Technology Shanghai 200040 China Institute of Medical Science Shanghai Jiao Tong University School of Medicine 280 South Chongqing Road Shanghai 200025 China

The Streptomyces phage φC31 integrase can efficiently target attB-bearing transgenes to endogenous pseudo attP sites within mammalian genomes. To better understand the activity of φC31 integrase in the bovine genome, DNA sequences of 44 integration events were analyzed, and 32 pseudo attP sites were identified. The majority of these sites share a sequence motif that contains inverted repeats and has similarities to wild-type attP site. Genomic DNA flanking these sites typically contained repetitive sequence elements, such as short and long interspersed repetitive elements. These sequence features indicate that DNA sequence recognition plays an important role in guiding φC31-mediated site-specific integration. In addition, BF27 integration hotspot sites were identified in the bovine genome, which accounted for 13.6% of all isolated integration events and mapped to an intron of the deleted in liver cancer 1 （DLC1） gene. Also we found that the pseudo attP sites in the bovine genome had other features in common with those in the human genome. This study represents the first time that the sequence features of pseudo attP sites specific integrase system has great potential for applied modifications in the bovine genome were analyzed. We conclude that this site- of the bovine genome.

关键词： φC31 integrase Pseudo attP sites Sequence motif Repetitive elements Bovine genome

DELAYED FLOWERING, an Arabidopsis Gene That Acts in the Autonomous Flowering Promotion Pathway and Is Required for Normal Development

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Journal of Integrative Plant Biology 2006年第1期48卷 27-34页

作者： Ming-Jie Chen Zheng Yuan Hai Huang Key Laboratory of Molecular genetics Shanghai Institute of Plant Physiology and Ecology Graduate School of the Chinese Academy of Sciences Shanghai 200032 China

The control of flowering time in higher plants is one of the most important physiological processes and is critical for their reproductive success. To investigate the mechanisms controlling flowering time, we screened for Arabidopsis mutants with late-flowering phenotypes. One mutant, designated delayed flowering （dfr） in the Landsberg erecta （Ler） ecotype, was identified with delayed flowering time. Genetic analysis revealed that dfr is a single gene recessive nuclear mutant and the mutation was mapped to a locus tightly linked to UFO on chromosome 1. To our knowledge, no gene regulating flowering time has been reported yet in this region. The dfrmutant plant showed a delayed flowering time under the different growth conditions examined, including long- and short-day photoperiods and gibberellic acid GA3 treatments, suggesting that DFR is a gene involved in the autonomous flowering promotion pathway. The Arabidopsis gene FLOWERING LOCUS C （FLC） plays a central role in repressing flowering and its transcripts are undetectable in wild-type Ler. However, FLCexpression was upregulated in the dfrmutant, suggesting that DFR is a negative regulator of FLC. In addition, the dfr mutant plant displayed altered valve shapes of the silique and the number of trichomes and branches of each trichome were both reduced, indicating that the DRFgene is also required for normal plant development. Moreover, dfr leafy-5 （Ify-5） double mutant plants showed a much later flowering time than either dfr or Ify-5 single mutants, indicating that DFR and LFY act synergistically to promote flowering in Arabidopsis.

关键词： Arabidopsis autonomous pathway dfr, flowering time trichome.

The forty years of medical genetics in China

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Journal of genetics and Genomics 2018年第11期45卷 569-582页

作者： Lei Cai Lan Alice Zheng Lin He Bio-X Institutes Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education)Collaborative Innovation Center for Genetics and DevelopmentShanghai Jiao Tong UniversityShanghai 200240China Shanghai Center for Women and Children's Health Shanghai 200062China Stanley Center Broad InstituteCambridgeMA 02142USA

Medical genetics is the newest cutting-edge discipline that focuses on solving medical problems using genetics knowledge and methods. In China, medical genetics research activities initiated from a poor inner basis but a prosperous outer environment. During the 40 years of reform and opening-up policy,Chinese scientists contributed significantly in the field of medical genetics, garnering considerable attention worldwide. In this review, we highlight the significant findings and/or results discovered by Chinese scientists in monogenic diseases, complex diseases, cancer, genetic diagnosis, as well as gene manipulation and gene therapy. Due to these achievements, China is widely recognized to be at the forefront of medical genetics research and development. However, the significant progress and development that has been achieved could not have been accomplished without sufficient funding and a wellconstructed logistics network. The successful implementation of translational and precise medicine sourced from medical genetics will depend on an open ethics policy and intellectual property protection,along with strong support at the national industry level.

关键词： Medical genetics GWAS CRISPR Cancer Monogenic diseases Complex diseases Genetic diagnosis Gene therapy

Establishing motor disorder mouse models of Parkinson disease Comparison of 6-hydroxydompamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

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Neural Regeneration Research 2007年第10期2卷 611-616页

作者： Zhi hua Ren Jie Gao Yan Chen Zhen yu Lu Department of Medical genetics Basic Medical CollegeShanghai JiaoTong UniversityShanghai200025China

At present, pathogenesis and mechanism of Parkinson disease （PD） are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histological characteristics and motor behavior dysfunction. In recent years, transgenic mice have been widely utilized to study the mechanism of PD, and it has become imperative that a PD mouse model of motor behavioral dysfunction be established. OBJECTIVE： To compare the behavioral and histochemical characters of two neurotoxic mice model induced with 6-hydroxydopamine （6-OHDA） or 1-methyl-4-phenyl-1, 2, 3, 6 -tetrahydropyridine （MPTP）, and a better method to mimic Parkinson disease will be found out. DESIGN： Parallel experiment. SETTING： Laboratory of Molecular genetics, Department of Medical genetics, Shanghai Jiao Tohg University. MATERIALS： Sixty 129Sv/C57BL6J male wild mice, SPF grade, 8 - 12 weeks old, weighing 20 - 25 g, were provided by Experimental Animal Center, Shanghai Jiao Tong University. All the surgery operation was performed according to the rules of Shanghai Jiaotong University Animal committee. METHODS： The experiment was carried out in the Laboratory of Molecular genetics （National Key Laboratory）, Department of Medical genetics, Shanghai Jiao Ttong University from March to August 2006. ①Thirty-two male mice were randomly divided into control group and drug treatment group with 16 mice in each group. Surgery was carried out and 6-OHDA was administrated to substantia nigra pars compacta （SNpc） and nigra-striatum pathway according to the different parameters with intoxication apparatus. Saline was injected to the other 16 mice according to the same paradigm. 1 mg/kg apomorphine was injected intraperitoneally 2 weeks later after surgery to induce the imbalanced rotation behavior for 40 minutes. ②Twenty-eight mice were randomly divided into 4 groups with 7 in each group, including low-dose, moderate-dose, high-dose groups and negative contr

关键词： Parkinson disease 6-hydroxydopamine 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine motor behavioral dysfunction

Development of T. aestivum ***. californicum Alien Chromosome Lines and Assignment of Homoeologous Groups of Hordeum californicum Chromosomes

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Journal of genetics and Genomics 2014年第8期41卷 439-447页

作者： Yuhui Fang Jingya Yuan Zhangjun Wang Haiyan Wang Jin Xiao Zhixi Yang Ruiqi Zhang Zengjun Qi Weigang Xu Lin Hu Xiu-E Wang State Key Laboratory of Crop genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University Wheat Research Center Henan Academy of Agricultural Sciences

Hordeum californicum （2n = 2x = 14, HH） is resistant to several wheat diseases and tolerant to lower nitrogen. In this study, a molecular karyotype of H. californicum chromosomes in the Triticum aestivum L. cv. Chinese Spring （CS）-H. californicum amphidiploid （2n = 6x = 56, AABBDDHH） was established. By genomic in situ hybridization （GISH） and multicolor fluorescent in situ hybridization （FISH） using repetitive DNA clones （pTa71, pTa794 and pSc119.2） as probes, the H. californicum chromosomes could be differentiated from each other and from the wheat chromosomes unequivocally. Based on molecular karyotype and marker analyses, 12 wheat--alien chromosome lines, including four disomic addition lines （DAH1, DAH3, DAH5 and DAH6）, five telosomic addition lines （MtH7L, MtHIS, MtH1L, DtH6S and DtH6L）, one multiple addition line involving H. californicum chromosome H2, one disomic substitution line （DSH4） and one translocation line （TH7S/1BL）, were identified from the progenies derived from the crosses of CS-H. californicum amphidiploid with common wheat varieties. A total of 482 EST （expressed sequence tag） or SSR （simple sequence repeat） markers specific for individual H. californicum chromosomes were identified, and 47, 50, 45, 49, 21, 51 and 40 markers were assigned to chromosomes H1, H2, H3, H4, H5, H6 and H7, respectively. According to the chromosome allocation of these markers, chromosomes H2, H3, H4, H5, and H7 of H. californicum have relationship with wheat homoeologous groups 5, 2, 6, 3, and 1, and hence could be designated as 5Hc, 2He, 6Hc, 3Hc and 1Hc, respectively. The chromosomes H1 and H6 were designated as 7Hc and 4Hc, respectively, by referring to SSR markers located on rye chromosomes.

关键词： Hordeum californicum Chinese Spring--H. californicum amphiploid GISH/FISH Alien chromosome lines Homoeologous group

Impact of umbelliprenin-containing niosome nanoparticles on VEGF-A and CTGF genes expression in retinal pigment epithelium cells

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International Journal of Ophthalmology(English edition) 2024年第1期17卷 7-15页

作者： Farzad Dastaviz Akram Vahidi Teymoor Khosravi Ayyoob Khosravi Mehdi Sheikh Arabi Abouzar Bagheri Mohsen Rashidi Morteza Oladnabi Stem Cell Research Center Golestan University of Medical SciencesGorgan ***Iran Department of Medical genetics School of Advanced Technologies in MedicineGolestan University of Medical SciencesGorgan ***Iran Student Research committee Golestan University of Medical SciencesGorgan ***Iran Department of Molecular Medicine Faculty of Advanced Medical TechnologiesGolestan University of Medical SciencesGorgan ***Iran Department of Medical Nanotechnology Faculty of Advanced Medical TechnologiesGolestan University of Medical SciencesGorgan ***Iran Department of Clinical Biochemistry and genetics Faculty of MedicineMazandaran University of Medical SciencesSari ***Iran Department Pharmacology Faculty of MedicineMazandaran University of Medical SciencesSari ***Iran Gorgan Congenital Malformations Research Center Golestan University of Medical SciencesGorgan ***Iran

AIM:To investigate the impact of niosome nanoparticles carrying umbelliprenin(UMB),an anti-angiogenic and anti-inflammatory plant compound,on the expression of vascular endothelial growth factor(VEGF-A)and connective tissue growth factor(CTGF)genes in a human retinal pigment epithelium(RPE)-like retina-derived cell ***:UMB-containing niosomes were created,optimized,and ***-like cells were treated with free UMB and UMB-containing *** IC_(50)values of the treatments were determined using an MTT *** expression of VEGF-A and CTGF was evaluated using real-time polymerase chain reaction after RNA extraction and cDNA ***’characteristics,including drug entrapment efficiency,size,dispersion index,and zeta potential were *** UMB had an IC_(50)of 96.2μg/mL,while UMB-containing niosomes had an IC_(50)of 25μg/***:Treatment with UMB-containing niosomes and free UMB resulted in a significant reduction in VEGF-A expression compared to control cells(P=0.001).Additionally,UMB-containing niosomes demonstrated a significant reduction in CTGF expression compared to control cells(P=0.05).However,there was no significant reduction in the expression of both genes in cells treated with free ***:Both free UMB and niosome-encapsulated UMB inhibits VEGF-A and CTGF genes ***,the latter demonstrates significantly greater efficacy,potentially due to the lower UMB dosage and gradual *** findings have implications for anti-angiogenesis therapeutic approaches targeting age-related macular degeneration.

关键词： umbelliprenin niosome age-related macular degeneration vascular endothelial growth factor-A connective tissue growth factor

Interaction between circulating cancer cells and platelets:clinical implication

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Chinese Journal of Cancer Research 2015年第5期27卷 450-460页

作者： Xiao-Liang Lou Jian Sun Shu-Qi Gong Xue-Feng Yu Rui Gong Huan Deng Molecular Medicine and genetics Center The Fourth Affiliated Hospital of Nanchang University Department of Pathology The Fourth Affiliated Hospital of Nanchang University Renmin Institute of Forensic Medicine

Metastasis is the main cause of cancer-associated mortality. During this complicated process, some cancer cells, also called circulating tumor cells （CTCs）, detach from primary sites, enter bloodstream and extravasate at metastatic site. Thrombocytosis is frequently observed in patients with metastatic cancers suggesting the important role of platelets in metastasis. Therefore this review focuses on how platelets facilitate the generation of CTCs, protect them from various host attacks, such as immune assaults, apoptosis and shear stress, and regulate CTCs intravasation/extravasation. Platelet-derived cytokines and receptors are involved in this cascade. Identification the mechanisms underlie platelet-CTCs interactions could lead to the development of new platelet-targeted therapeutic strategy to reduce metastasis.

关键词： Circulating tumor cells (CTCs) platelet epithelial-mesenchymal transition (EMT) immunesurveillance metastasis

Elevational patterns of bird species richness on the eastern slope of ***,Sichuan Province,China

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Avian Research 2019年第1期10卷 1-12页

作者： Xingcheng He Xiaoyi Wang Shane DuBay Andrew Hart Reeve Per Alstrom Jianghong Ran Qiao Liu Yongjie Wu Key Laboratory of Bio-resources and Eco-environment of Ministry of Education College of Life SciencesSichuan University committee on Evolutionary Biology University of Chicago Life Sciences Section Integrative Research CenterField Museum of Natural History Biosystematics Section Natural History Museum of DenmarkUniversity of Copenhagen Department of Ecology and genetics Animal EcologyEvolutionary Biology CentreUppsala University Swedish Species Information Centre Swedish University of Agricultural Sciences Key Laboratory of Zoological Systematics and Evolution Institute of ZoologyChinese Academy of Sciences Institute of Mountain Hazards and Environment Chinese Academy of Sciences

Background:In biological systems,biological diversity often displays a rapid turn-over across *** defining feature has made mountains classic systems for studying the spatial variation in *** patterns of elevational diversity can vary among lineages and mountain systems it remains difficult to extrapolate findings from one montane region to another,or among *** this study,we assessed patterns and drivers of avian diversity along an elevational gradient on the eastern slope of ***,the highest peak in the Hengduan Mountain Range in central China,and a mountain where comprehensive studies of avian diversity are still ***:We surveyed bird species in eight 400-m elevational bands from 1200 to 4400 m *** 2012 and *** test the relationship between bird species richness and environmental factors,we examined the relative importance of seven ecological variables on breeding season distribution patterns:land area(LA),mean daily temperature(MDT),seasonal temperature range(STR),the mid-domain effect(MDE),seasonal precipitation(SP),invertebrate biomass(IB) and enhanced vegetation index(EVI).Climate data were obtained from five local meteorological stations and three temperature/relative humidity smart sensors in ***:A total of 219 bird species were recorded in the field,of which 204 were recorded during the breeding season(April–August).Species richness curves(calculated separately for total species,large-ranged species,and smallranged species) were all ***-ranged species contributed more to the total species richness pattern than small-ranged *** and IB were positively correlated with total species richness and small-ranged species *** and MDT were positively correlated with small-ranged species richness,while STR and SP were negatively correlated with small-ranged species *** was positively correlated with large-ranged species *** we considered the combina

关键词： Mt.Gongga Elevational gradient Environmental factors Species richness Stable microclimate