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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 644-648页

作者： Peng-Da MA1, Tian-Cheng LU1, Xiao-Fu ZHOU1,2, Xiao-Juan ZHU1, and Xing-Zhi WANG1*1The institute of genetics and Cytology, Northeast normal university, Changchun 130024, china 2.he institute of gene technology, jilin normal university, siping 13.000, china The institute of genetics and Cytology Northeast Normal University The institute of gene technology Jilin Normal University

Spinach Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) large (rbcL) and small (rbcS) subunits were separated by SDS-PAGE, and protein amount and purity were determined by Bradford assay. Polyclonal antibodies against rbcL and rbcS subunit were generated in female BALB/c mice and had no cross-reaction with each other. A total of 81 μg antigens were used and 0.3 ml anti-sera with titer of 1:5000 were yielded. The antibodies were also applicable to study rbcL and rbcS in tobacco plant Nicotiana benthamiana. Potato virus X vector pGR107 induced silencing of rbcS gene by Agrobacterium in Nicotiana benthamiana was performed. The expression level of rbcL and rbcS was lower in rbcS silenced plants than that in control plants as detected by the corresponding antibodies. This implied that the expression of rbcL was regulated by rbcS.

关键词： ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) rbcL rbcS polyclonal antibody virus-induced gene silencing (VIGS)

Structural components of the nuclear body in nuclei of Allium cepa cells

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Cell Research 2001年第1期11卷 68-73页

作者： TAO WEI, CHANG HUI YAN, TAO CAI, SHUI HAO, ZHONG he ZHAI (College of Life Sciences, Peking university, Beijing 100871, china) (institute of genetics and Cytology, Northeast normal university, Changchun jilin 130024, china) 1College of Life Sciences Peking University Beijing 100871 China 1College of Life Sciences Peking University Beijing 100871 China 1College of Life Sciences Peking University Beijing 100871 China 2institute of genetics and Cytology Northeast Normal University Changchun Jilin 130024 China 1College of Life Sciences Peking University Beijing 100871 China

Nuclear bodies have long been noted in interphase nuclei of plant cells, but their structural component, origin and fUnction are still unclear by now. The present work showed in onion cells the nuclear bodies appeared as a spherical structure about 0.3 to 0.8 μm in diameter. They possibly were formed in nucleolus and subsequently released, and entered into nucleoplasm. Observation through cytochemical staining method at the ultrastructural level codeined that nuclear bodies consisted of ribonucleoproteins (RNPs) and silver-stainable proteins. Immunocytochemical results revealed that nuclear bodies contained no DNA and ribosomal gene transcription factor (UBF). Based on these data, we suggested that nuclear bodies are not related to the ribosome or other gene transcription activities, instead they may act as subnuclear structures for RNPs transport from nucleolus to cytoplasm, and may also be involved in splicing of pre-mRNAs.

关键词： Nuclear body structural component ribonucleoproteins nucleolus Allium cepa.

A PSTAIRE CDK-like protein localizes in nuclei and cytoplasm of Physarum polycephalum and functions in the mitosis

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Cell Research 2004年第2期14卷 169-175页

作者： Gui Ying LI~(1,2), Miao XING~(1,*), Bo HU~(1,3) ~1College of Life Science, Shenzhen Key Laboratory of Microbial gene Engineering, Shenzhen university, Shenzhen 518060, china. ~2Department of Immunology, School of Basic Medical Sciences, jilin university, Changchun 130021, china ~3institute of genetics and Cytology, Northeast normal university, Changchun 130024, china. College of Life Science Shenzhen Key Laboratory of Microbial Gene Engineering Shenzhen University Shenzhen 518060 China.

CDKs play key roles in controlling cell cycle progression in all eukaryotes. In plants, multiple CDKs are present, among which the best characterized CDKs are PSTAIRE CDKs. In this study, we carried out Western blot, immunoelectron microscopy and antibody treatment with an anti-PSTAIRE monoclonal antibody to explore the subcellular localization and functions of PSTAIRE CDKs in Physarum polycephalum. The results of Western blot and immunoelectron microscopy showed that in P. polycephalum, a PSTAIRE CDK-like protein was 34 kD in molecular weight and located in both nuclei and cytoplasm. In nuclei, the protein was mainly associated with chromosomes and nucleoli. The expression of the PSTAIRE CDK-like protein in both the plasmodia and nuclei showed little fluctuation through the whole cell cycle. When treated with an anti-PSTAIRE monoclonal antibody at early S phase, the cells were arrested in S phase, and the mitotic onset of P. polycephalum was blocked for about 1 h when treated at early G2 phase. Our dat

关键词： PSTAIRE CDK-like protein Physarum polycephalum mitosis immunoelectron microscopy.

OIP5 Interacts with NCK2 to Mediate Human Spermatogonial Stem Cell Self-Renewal and Apoptosis through Cell Cyclins and Cycle Progression and Its Abnormality Is Correlated with Male Infertility

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Research 2024年第1期2023卷 455-469页

作者： Yinghong Cui Wei Chen Li Du Zuping he The Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province The Research Center of Reproduction and Translational Medicine of Hunan ProvinceHunan Normal University School of MedicineChangsha 410013HunanChina

Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative ***,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remain ***,we have demonstrated for the first time that OIP5(Opa interacting protein 5)controlled the self-renewal and apoptosis of human *** sequencing identified that NCK2 was a target for OIP5 in human SSCs,and interestingly,OIP5 could interact with NCK2 as shown by Co-IP(co-immunoprecipitation),IP-MS(mass spectrometry),and GST pulldown ***2 silencing decreased human SSC proliferation and DNA synthesis but enhanced their ***,NCK2 knockdown reversed the influence of OIP5 overexpression on human ***,OIP5 inhibition decreased the numbers of human SSCs at S and G2/M phases,while the levels of numerous cell cycle proteins,including cyclins A2,B1,D1,E1 and H,especially cyclin D1,were remarkably ***,whole-exome sequencing of 777 patients with nonobstructive azoospermia(NOA)revealed 54 singlenucleotide polymorphism mutations of the OIP5 gene(6.95%),while the level of OIP5 protein was obviously lower in testes of NOA patients compared to fertile ***,these results implicate that OIP5 interacts with NCK2 to modulate human SSC self-renewal and apoptosis via cell cyclins and cell cycle progression and that its mutation and/or lower expression is correlated with *** such,this study offers novel insights into molecular mechanisms underlying the fate determinations of human SSCs and the pathogenesis of NOA,and it provides new targets for treating male infertility.

关键词： OIP5 Apoptosis reversed

Identification and Validation of Vascular-Associated Biomarkers for the Prognosis and Potential Pathogenesis of Hypertension Using Comprehensive Bioinformatics Methods

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World Journal of Cardiovascular Diseases 2024年第3期14卷 115-128页

作者： Xiangguang Chang Lei Guo Liying Zou Yazhao Ma jilin Feng Department of Laboratory Yangbi Yi Autonomous County People’s Hospital Dali China Department of Obstetrics and Gynecology Yangbi Yi Autonomous County People’s Hospital Dali China Department of Infectious Diseases Yangbi Yi Autonomous County People’s Hospital Dali China Medical Community Office Yangbi Yi Autonomous County People’s Hospital Dali China

Background: Hypertension, also known as increased blood pressure, is a phenomenon in which blood flows in blood vessels and causes persistently higher-than-normal pressure on the vessel wall. The identification of novel prognostic and pathogenesis biomarkers plays a key role in the management of hypertension. Methods: The GSE7483 and GSE75815 datasets from the gene expression omnibus (GEO) database were used to identify the genes associated with hypertension that were differentially expressed genes (DEGs). The functional role of the DEGs was elucidated by gene body (GO) enrichment analysis. In addition, we performed an immune infiltration assay and GSEA on the DEGs of hypertensive patients and verified the expression of novel DEGs in the blood of hypertensive patients by RT-qPCR. Results: A total of 267 DEGs were identified from the GEO database. GO analysis revealed that these genes were associated mainly with biological processes such as fibroblast proliferation, cell structural organization, extracellular matrix organization, vasculature development regulation, and angiogenesis. We identified five possible biomarkers, Ecm1, Sparc, Sphk1, Thbsl, and Mecp2, which correlate with vascular development and angiogenesis characteristic of hypertension by bioinformatics, and explored the clinical expression levels of these genes by RT-qPCR, and found that Sparc, Sphk1, and Thbs1 showed significant up-regulation, in agreement with the results of the bioinformatics analysis. Conclusion: Our study suggested that Sparc, Sphk1 and Thbs1 may be potential novel biomarkers for the diagnosis, treatment and prognosis of hypertension and that they are involved in the regulation of vascular development and angiogenesis in hypertension.

关键词： Hypertension Biomarkers Differentially Expressed genes Vascular Development and Angiogenesis Bioinformatics Analysis

Culture of Bovine Fetal Skin Fibroblasts in vitro and Ipr1 gene Transfection

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畜牧兽医学报 2011年第S1期 7-12页

作者： SONG Yong-li 1,2.he Xiao-ning 2,HUA Song 2,LAN Jie 2,ZheNG Yue-mao 2,LI Ji-xia 2.he Xiao-ying 2,QUAN Fu-sheng 2,ZHANG Yong 2(1. College of Veterinary Medicine,jilin Agricultural Science and technology College,jilin132101,china 2. Key Laboratory of Animal Reproductive Physiology & Embryo technology,Ministry of Agriculture,College of Veterinary Medicine,Northwest Agricuture and Forestry university,Yangling 712100,china)

In order to generate transgenic donor cells for nuclear transfer, bovine fetal fibroblasts were isolated in vitro and transfected with the eukaryotic expression vector pSRA-EGFP-Ipr1. The mouse Ipr1 gene and human SR-A promoter were successfully cloned and then used to construct this macrophage-specific eukaryotic expression vector. Bovine fetal fibroblasts in stable primary culture (4th passage) were transfected with pSRA-EGFP-Ipr1 by electroporation. Fluorescence from GFP was observed after 24h. Transgenic cells were selected using G418 and the resultant monoclones were picked and expanded. The transgenic cells, at the 9 th passage, were evaluated by PCR and flow cytometry. The inserted Ipr1 was confirmed by PCR, indicating stable integration of the transgene into the genome and cells had normal karyotypes and very good appearance, which indicate no deleterious result of the transgenesis. In conclusion, the cells obtained could be used as donor cells for nuclear transfer for further research of transgenic cattle.

关键词： donor cells eunuclear expression vector Ipr1 gene bovine fetal fibroblasts cells electroporation EGFP

Expression of Survivin gene among Human normal Endometrium,Atypical Hyperplasia of Endometrium and Endometrial Carcinoma

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Journal of Reproduction and Contraception 2010年第3期21卷 129-135页

作者： Jian-ying XU Shu-fang WANG Zhong-yan WAN Wei-jie ZHU institute of Reproductive Immunology Jinan University Guangzhou 510632 China Department of Obstetrics and Gynecology the Second People Hospital of Zhuhai Zhuhai 519000 China

Objective To investigate the expression of survivin gene among human normal endometrium,atypical hyperplasia of endometrium,and endometrial *** Tissue samples of human normal endometrium during proliferative phase （NE,n=20）,atypical hyperplasia of endometrium （Ahe,n=11）,and endometrial carcinoma （EC,n=9） were ***,Paraffin embedded sections of NE （n=20）,Ahe （n=20）,and EC （n=20） were *** expression of survivin gene was determined by immunohistochemistry and the real-time reverse transcription polymerase chain *** Survivin immunostaining appeared in the cytoplasm of endometrial epithelial *** survivin staining and mRNA had higher levels in Ahe or EC than those in NE （P〈0.01）.No difference was found on survivin staining and mRNA between Ahe and EC （P〉0.05）.Conclusion High expression of survivin gene in human endometrium is associated with the risk of atypical hyperplasia progressing to endometrial *** high level of survivin expression is useful as a predictive indicator for endometrial carcinoma.

关键词： survivin endometrium atypical hyperplasia of endometrium endometrial carcinoma

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Phylogenetic relationships of Cyprinidae (Teleostei:Cypriniformes) inferred from the partial S6K1 gene sequences and implication of indel sites in intron 1

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Science china(Life Sciences) 2007年第6期50卷 780-788页

作者： KONG XiangHui1,2, WANG XuZhen1, GAN XiaoNi1, LI JunBing1 & he ShunPing1 1 institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, china 2 College of Life Science, henan normal university, Xinxiang 453007, china institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China College of Life Science Henan Normal University Xinxiang 453007 China

The family Cyprinidae is widely distributed in East Asia,; has the important phylogenetic significance in the fish evolution. In this study, the 5′ end partial sequences (containing exon 1, exon 2; indel 1) of S6K1 gene were obtained from 30 representative species in Cyprinidae; outgroup using PCR amplification; sequencing. The phylogenetic relationships of Cyprinidae were reconstructed with neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML),; Bayesian methods. Myxocyprinus asiaticus (Catostomidae) was assigned to the outgroup taxon. Similar phylogenetic relationships within the family Cyprinidae were achieved with the four analyses. Leuciscini; Barbini were monophyletic lineages respectively with the high nodal supports. Leuciscini comprises Hypophthalmichthyinae, Xenocyprinae, Cultrinae, Gobioninae, Acheilognathinae; East Asian species of Leuciscinae; Danioninae. Monophyly of East Asian clade was supported with high nodal support. Barbini comprises Schizothoracinae, Barbinae, Cyprininae; Labeoninae. The monophyletic lineage consisting of Danio rerio, D. myersi,; Rasbora trilineata was basal in the tree. In addition, the large fragment indels in intron 1 were analyzed to improve the understanding of Cyprinidae relationships. The results showed that the large fragment indels were correlated with the relations among species. Some conserved regions in intron 1 were thought to be involved in the functional regulation. However, no correlation was found between sequence variations; species characteristic size.

关键词： Cyprinidae, S6K1 gene, phylogenetic relationships, intron

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Hairy roots induced by Agrobacterium rhizogenes and production of regenerative plants in hairy root cultures in maize

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Science china Life Sciences 2006年第4期 305-310页

作者： XU Hongwei1,2*, ZHOU Xiaofu2,3*, LU Jingmei1, WANG Junjie1 & WANG Xingzhi1 1. School of Life Sciences, Northeast normal university, Changchun 130024, china 2. institute of gene Engineering, jilin normal university, siping 13.000, china

Hairy roots of maize were induced by infecting 15-d calli with Agrobacterium rhizogenes. The hairy roots cultured in hormone-free media showed the vigorous growth and typical hairy root features. The regenerated plants were produced from hairy roots in MS media supplemented with 1.6 mg/L ZT and 0.4 mg/L NAA. The PCR-Southern hybridization demonstrated that T-DNA had been integrated into the chromosome of regenerated plants.

关键词： Agrobacterium rhizogenes maize hairy root PCR-Southern.

Molecular evolution of connective tissue growth factor in Cyprinidae (Teleostei:Cypriniformes)

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Progress in Natural Science:Materials International 2008年第2期18卷 155-160页

作者： Xianghui Kong a,b,Xuzhen Wang a,Xiaoni Gan a,Junbing Li a,Shunping he a,a institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,china b College of Life Science,henan normal university,Xinxiang 453007,china institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China College of Life Science Henan Normal University Xinxiang 453007 China

Connective tissue growth factor (CTGF) plays an important role in regulation of cell growth,differentiation,apoptosis and individual development in animals. The study of sequences variation and molecular evolution of CTGF gene across various species of the cyprinid could be helpful for understanding of speciation and gene divergence in this kind of fish. In this study,19 novel sequences of CTGF gene were obtained from the representative species of the family Cyprinidae using PCR amplification,cloning and sequencing. Phylogenetic relationships of Cyprinidae were reconstructed by neighbor-joining (NJ),maximum parsimony (MP),maximum likelihood (ML) and Bayesian method. Oryzias latipes from the family Cyprinodontidae was assigned to be the outgroup taxon. Leuciscini and Barbini were clustered into the monophyletic lineages,respectively,with the high nodal supports. The estimation of the ratio of non-synonymous to synonymous substitution (dN/dS) for the various branches indicated that there stood the different evolution rates between the Leuciscini and the Barbini. With the ratio of dN/dS of the Leuciscini being lower than that of the Barbini,species within the Barbini were demonstrated to be subjected to the relatively less selection pressure and under the relaxable evolution background. A 6 bp indel (insertion/deletion) was found at the 50 end of CTGF gene of Cyprinidae,and this 6 bp deletion only appeared in the Leuciscini,which is a typical characteristic of the Leuciscini and provides evidence for the monophylogeny of the Leuciscini. For the amino acid sequences of CTGF protein,the most variations and indels were distributed in the signal region and IGFBP region of this protein,implying that these variations were correlated with the regulation of the CTGF gene expression and protein activity.

关键词： Connective tissue growth factor Sequence variation Molecular evolution Cyprinidae