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expression of some tumor associated factors in human carcinogenesis and development of gastric carcinoma

World Journal of Gastroenterology 2005年第21期11卷 3217-3221页

作者： Ming-DongZhao Xue-MeiHu Dian-JingSun QunZhang Yu-HaoZhang WeiMeng DepartmentofRadiology AffiliatedHospitalofBinzhouMedicalCollegeBinzhou256603ShandongProvinceChina DepartmentofImmunologyofBinzhouMedicalCollege Binzhou256603ShandongProvinceChina MedicalSchoolofShanghaiJiaotongUniversity Shanghai200030China

AIM: To study the effect of IGF-1/IGF-1R and gastrin/CCK-BR on carcinogenesis and development of human gastric carcinoma and to explore its mechanism and provide a credible theoretical foundation for early diagnosis and molecular therapy of gastric ***: mRNA expression levels of IGF-1/IGF-1R and gastrin/CCK-BR were assessed by RT-PCR method in gastric cancer tissues, adjacent mucosa, and tumor-free tissues from 56 patients with gastric carcinoma and normal gastric mucosae from 56 healthy controls. Tissue specimens were obtained by biopsy and confirmed by histological ***: The mRNA levels of IGF-1/IGF-1R were increased in gastric cancer tissues compared with normal tissues from healthy controls and successively increased in tumor-free tissues, adjacent mucosa, and gastric cancer tissues. The mRNA levels of gastrin/CCK-BR were increased in gastric cancer tissues compared with normal tissues from healthy controls. There was a significant difference between gastric cancer tissues and adjacent mucosa and tumor-free tissues, but the mRNA levels of gastrin were not significantly increased in adjacent mucosa and gastric cancer tissues compared with tumorfree tissues. The mRNA levels of CCK-BR were increased in gastric cancer tissues and adjacent mucosa compared with tumor-free tissues, but not significantly increased in adjacent mucosa and gastric cancer tissues compared with gastric cancer ***: Overexpression of IGF-1/IGF-1R and gastrin/CCK-BR promotes the disorderly proliferation of gastric mucosa epithelia and it is of great significance in the carcinogenesis and development of gastric carcinoma.

关键词： Gastric carcinoma mRNA IGF-1 IGF-1R Gastrin CCK-BR protein expression

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expression and Purification of ZNF191（243-368） in Three expression Systems

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Chinese Journal of Chemistry 2007年第11期25卷 1739-1742页

作者：赵东欣滕欣辰丁志春黄仲贤 Department of Chemistry Fudan University Shanghai 200433 China School of Chemistry and Chemical Engineering Henan University of Technology Zhengzhou Henan 450001 China

ZNF191 （243-368）, a new human zinc finger protein, probably relates to some hereditary diseases and cancers, To obtain adequate amount of ZNF191（243-368） for the study of its property, structure and function, three different expression systems of inclusion-body, glutathione S-transferase （GST）, and hexahistidine （6 × His） were used and compared. Among these systems, the expression level of ZNFI91（243-368） was increased in inclusion body system under a higher isopropylthio-β-D-galactoside （IPTG） concentration, but the non-target proteins were also increased more, which made its purification more difficult and the yield lower. The expression of His-tag fusion protein was almost not affected by IPTG concentration, temperature and inducing time. At a high IPTG concentration the highest expression yield for GST fusion protein was obtained. And the fusion proteins can be partially purified by a single affinity chromatography step. The fusion protein systems show advantages for expression of these proteins.

关键词： protein expression affinity chromatography fusion protein isopropylthio-β-D-galactoside

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Hepatoma cells up-regulate expression of programmed cell death-1 on T cells

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World Journal of Gastroenterology 2008年第44期14卷 6853-6857页

作者： Ji Chen Xue-Jie Wu Gui-Qiang Wang Department of Infectious Diseases & Center for Liver Diseases Peking University First Hospital Beijing 10034 China

AIM： To investigate the effect of hepatoma cells on up-regulation of programmed cell death-1 （PD-1）, and the function of PD-1 on T cells. METHODS： HepG2 or HepG2.***.1.5 cells were cocultured with a lymphoma cell line-Jurkat cells. PD-1 expression was detected by flow cytometry. IL：2, INF-γ and IL-10 in culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Cytotoxic action of T cells was determined by MIF reduction assay-direct mononuclear cell cytotoxicity assay. RESULTS： The PD-1 expression on Jurkat cells increased by 16.17% ± 2.5% and 17.43% ± 2.2% after HepG2 or HepG2.***.1.5 cells were co-cultured for 48 h. The levels of IL-2, INF-γ and IL-10 in the culture supernatant were 202.9 ＋ 53.0 pg/mL, 88.6 ± 4.6 pg/mL and 63.7± 13.4 pg/mL respectively, which were significantly higher than those （102.9 ± 53 pg/mL, 39.3 ± 4.2 pg/mL, and 34.6 =E13.7 pg/mL） in the control group （P 〈 0.05）. The OD value for MTT assay in the blocking group （0.29 ± 0.06） was significantly higher than that （0.19 ± 0.09） in the control group （P 〈 0.05）. CONCLUSION： PD-1 expression on Jurkat cells is upregulated by hepatoma cells, cytokines and cytotoxic action are elevated after PD-1/PD-L1 is blocked.

关键词： Hepatoma cell Programmed cell death-l protein expression T cell function Cytokine

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Clinical signifi cance of FABP2 expression in newborns with necrotizing enterocolitis

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World Journal of Pediatrics 2016年第2期12卷 159-165页

作者： Yu Liu Liang-Fu Jiang Rong-Peng Zhang Wen-Tong Zhang Department of Pediatric Surgery Qilu HospitalShandong UniveristyJi'nanChina Department of Pediatric Surgery Linyi People's Hospital affi liated to Shandong UniveristyLinyiChina

Background: This meta-analysis aimed to determinethe role of human fatty acid binding protein 2 (FABP2)expression in the diagnosis of necrotizing enterocolitis(NEC) of *** sources: Eligible studies for further statisticalanalysis were identified from various databases including PubMed, Expert Medica Database, Web of Science,Cochrane Library, Google Scholar, China BioMedicineand China National Knowledge Infrastructure. Randomeffects model was used, and summary standardized meandifference (SMD) with its 95% confi dence interval (CI) wascalculated to assess the association of FABP2 expressionand ***: Ten articles which included 572 infants (262infants with NEC and 310 healthy controls) were includedin the current meta-analysis. FABP2 showed a positiverelationship with NEC of newborns (SMD=2.88, 95%CI=2.09-3.67, Pexpression washigher in patients with advanced stage of NEC (stage IIIor stage II+III) than in those with early stage of NEC(stage I) (SMD=-0.48, 95% CI=-0.87 to -0.09, P=0.015).Ethnicity-stratifi ed analysis yielded signifi cantly differentestimates with a high FABP2 expression in NEC in bothCaucasians (SMD=3.16, 95% CI=1.90-4.43, Pexpressionwas positively correlated with neonatal NEC in bothurinary- and blood-sample subgroups (all Pexpression is related to the damage to intestinal cells,which may be a possible early detection marker identifyingneonatal NEC.

关键词： fatty acid binding protein 2 meta-analysis necrotizing enterocolitis of newborn protein expression

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expression of Arginyl-tRNA Synthetase in Rats with Focal Cerebral Ischemia

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2014年第2期34卷 172-175页

作者：符荣范云智范宇葱赵洪洋 Department of Neurosurgery Union Hospital Tongji Medical College Huazhong University of Science and Technology

Aminoacyl-tRNA syntheses （AARS） can catalyze the adenosine triphosphate （ATP）-dependent acylation of their cognate tRNA（s） with a specific amino acid. They can be seen as an index to reflect the energy metabolic rate of ischemic brain cells in ischemic penumbra. This study ex- amined the relationship between arginyl-tRNA synthetase （ArgRS）, one of the AARS, and cerebral ischemia in rats. The model of middle cerebral artery occlusion （MCAO） was established in rats. The expression levels of ArgRS protein and mRNA were detected in rat brain tissues at different time points following MCAO by Western blotting and RT-PCR, respectively. The results showed that the MCAO model was successfully established. Western blotting and RT-PCR analysis revealed that the ArgRS protein and mRNA were expressed in brain cells in both ischemic and normal penumbra tissues. The expression levels of ArgRS protein and mRNA peaked at 6 h after MCAO and decreased gradually. At 24 h, the expression levels of ArgRs protein and mRNA in ischemic penumbral tissues were lower than those in normal tissues. The expression levels of ArgRS mRNA and protein in ischemic penumbra var- ied with ischemic time, suggesting that the energy metabolism of brain cells in penumbra changed dy- namically after ischemia to ensure the endogenous self-protection of the body. The brain oxygen supply should be improved as soon as possible, especially within 6-12 h after ischemia, so as to meet the de- mand for energy metabolism in ischemic penumbra and make sure the cell structure remains stable.

关键词： stroke cerebral ischemia arginyl-tRNA synthetase gene expression protein expression

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Preparation of rabbit anti-rat LRRN3 polyclonal antibody and study of its expression

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Neural Regeneration Research 2010年第13期5卷 1030-1034页

作者： Jing Yang Yanjin Wang Yan Cai Zhiying Yang Ganqiu Wu Jianguo Wen Xuegang Luo He Huang Department of Histology and Embryology Xiangya School of Medicine Central South University Changsha 410013 Hunan Province China Department of Human Anatomy and Neurobiology Xiangya School of Medicine Central South University Changsha 410013 Hunan Province China Department of Neurosurgery Xiangya Hospital Central South University Changsha 410013 Hunan Province China

BACKGROUND： Studies have shown that the Repeat superfamily, could be related to neural LRRN3, a member of the Neuron Leucine-Rich development, differentiation, information transmission, and other functions, but most studies have focused on nucleic acid levels and few have reported on LRRN3 protein levels. OBJECTIVE： To prepare rabbit anti-rat LRRN3 polyclonal antibody and to observe protein tissue expression profiles. DESIGN, TIME AND SEI-rlNG： In vitro, molecular, biological experiments were performed from October 2007 to April 2009 in Laboratory of Neurobiology at Xiangya School of Medicine, Central South University. MATERIALS： Immunization antigen, namely rat MaI-LRRN3C-His recombinant protein, was provided by the Laboratory of Neurobiology at Xiangya School of Medicine, Central South University. METHODS： Rat Mal-LRRN3C-His recombinant protein was used to immunize male, New Zealand rabbits, and rabbit anti-rat LRRN3 polyclonal antibody was prepared. MAIN OUTCOME MEASURES： Antibody purification was conducted using protein A affinity chromatography, and the LRRN3 anti-serum titer was identified using enzyme-linked immunosorbent assay. Immunohistochemical techniques and Western blot preliminary tests were used to determine LRRN3 protein expression profiles in adult rats. RESULTS： A highly purified rabbit anti-rat LRRN3 polyclonal antibody was obtained. Western Blot results from rat brain total protein revealed a band at 79 kD, which was consistent with the size of LRRN3. Immunohistochemistry results showed that protein was mainly expressed in the central nervous system, and no significant positive signals were observed in other tissues. Positive cells included neurons of cerebral cortex and hippocampal dentate gyrus granule cell layer, and cerebellar Purkinje cells. There was no positive expression in glial cells. CONCLUSION： Rabbit anti-rat LRRN3 polyclonal antibody was successfully prepared at a high purity from the prokaryotic-expressed MaI-LRRN3C-His recombinant

关键词： polyclonal antibody preparation protein expression LRRN3 developmental neurobiology neural regeneration

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Genetic Modification of Baculovirus expression Vectors

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Virologica Sinica 2012年第2期27卷 71-82页

作者： Shu-fen Li Hua-lin Wang Zhi-hong Hu Fei Deng State Key Laboratory of Virology and Joint-Lab of Invertebrate Virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China

As a protein expression vector, the baculovirus demonstrates many advantages over other vectors. With the development of biotechnology, baculoviral vectors have been genetically modified to facilitate high level expression of heterologous proteins in both insect and mammalian cells. These modifications include utilization of different promoters and signal peptides, deletion or replacement of viral genes for increasing protein secretion, integration of polycistronic expression cassette for producing protein complexes, and baculovirus pseudotyping, promoter accommodation or surface display for enhancing mammalian cell targeting gene delivery. This review summarizes the development and the current state of art of the baculovirus expression system. Further development of baculovirus expression systems will make them even more feasible and accessible for advanced applications.

关键词： Baculovirus protein expression Promoters Signal peptides Gene delivery

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Induction of intestinal metaplasia in stomach of dogs and expression of tumor-related proteins in animal gastric mucosa lesions

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Chinese Medical Journal 2000年第4期 48-51页

作者：王东旭房殿春刘为纹

Objective To observe the development and progression of intestinal metaplasia (IM) in dog's stomach and expression of tumor related proteins in gastric mucosa lesions Methods IM animal model was induced in stomach of Beagle dog by combining treatment of oral administration of N methyl N' nito N nitrosoguanidine (MNNG) and ranitidine (R) with X ray irradiation to the target organ expression of APC, p53, K ras and bcl 2 gene proteins in animal gastric mucosa lesions were determined with immunohistochemical method Results IM animal model was successfully induced and the dynamic pathological changes were observed by means of this model It was confirmed that the progression from normal epithelial cells to IM cells may require several stages, including superficial gastritis, chronic atrophic gastritis, slight focal IM and moderate or severe IM Aberrant bcl 2 protein can be detected in the atrophic mucosa epithelium and the abnormal expression of APC, K ras and bcl 2 can be found in IM of mucosa Conclusions IM model in stomach of Beagle dog can be successfully induced by our method (MNNG+R+ X ray) The progression from normal to IM in dog resembles that of human being and the expression of tumor related proteins (APC, bcl 2, K ras) may play a role in the malignant transformation of IM

关键词： Beagle dog intestinal metaplasia animal model tumor related gene protein expression

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Graphene quantum dots rescue protein dysregulation of pancreatic β-cells exposed to human islet amyloid polypeptide

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Nano Research 2019年第11期12卷 2827-2834页

作者： Ava Faridi Yunxiang Sun Monika Mortimer Ritchlynn R.Aranha Aparna Nandakumar Yuhuan Li Ibrahim Javed Aleksandr Kakinen Qingqing Fan Anthony W.Purcell Thomas P.Davis Feng Ding Pouya Faridi Pu Chun Ke ARC Centre of Excellence in Convergent Bio-Nano Science and Technology Monash Institute of Pharmaceutical SciencesMonash University381 Royal ParadeParkvilleVIC3052Australia Department of Physics Ningbo UniversityNingbo315211China Department of Physics and Astronomy Clemson UniversityClemsonSC29634USA Institute of Environmental and Health Sciences College of Quality and Safety EngineeringChina Jiliang UniversityHangzhou310018China Infection and Immunity Program&Department of Biochemistry and Molecular Biology Biomedicine Discovery InstituteMonash UniversityClaytonVictoria3800Australia Australian Institute for Bioengineering and Nanotechnology The University of QueenslandBrisbaneQld4072Australia

The amyloid aggregation of peptides and proteins is a hallmark of neurological disorders and type 2 *** islet amyloid polypeptide(IAPP),co-secreted with insulin by pancreaticβ-cells,plays dual roles in both glycemic control and the pathology of type 2 *** IAPP can activate the NLRP3 inflammasome and modulate cellular autophagy,apoptosis and extracellular matrix metabolism,no data is available concerning intracellular protein expression upon exposure to the *** surprisingly,how intracellular protein expression is modulated by nanoparticle inhibitors of protein aggregation remains entirely *** this study,we first examined the changing proteomes ofβTC6,a pancreaticβ-cell line,upon exposure to monomeric,oligomeric and fibrillar IAPP,and detailed cellular protein expression rescued by graphene quantum dots(GQDs),an IAPP *** found that 29 proteins were significantly dysregulated by the IAPP species,while majority of these proteins were nucleotide-binding ***,our liquid chromatography tandem-mass spectrometry,fluorescence quenching,helium ion microscopy,cytotoxicity and discreet molecular dynamics simulations data revealed a remarkable capacity of GQDs in regulating aberrant protein expression through H-bonding and hydrophobic interactions,pointing to nanomedicine as a new frontier against human amyloid diseases.

关键词： islet amyloid polypeptide(IAPP) oligomer amyloid protein expression graphene quantum dot

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Encouraging Solution to the Problem of Synthesizing protein α-Thioester

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Chinese Journal of Chemistry 2024年第10期42卷 1114-1120页

作者： Xinliang Liu Zijun Gao Jie Zhao Farong Ye Ping Huang Ping Wang Center for Chemical Glycobiology Shanghai Key Laboratory for Molecular Engineering of Chiral DrugsFrontiers Science Center for Transformative MoleculesSchool of Chemistry and Chemical EngineeringZhangjiang Institute for Advanced StudyShanghai JiaoTongUniversityShanghai 200240China Shenzhen Research Institute of Shanghai Jiao Tong University ShenzhenGuangdong 518057China

Expressed protein ligation(EPL)provides a powerful tool to access large-size proteins with precise *** methods for constructing the critical protein thioester for EPL have predominantly relied on the recombinant intein fusion expressed in Escherichia coli(***).Despite its powerful applications,the expression of thioester derived from eukaryotic protein in *** inherently suffers from its limited solubility,the inactivity of intein,premature hydrolysis and low *** overcome these obstacles,we present herein the facile one-flask synthesis of inaccessible proteinα-thioester via a SUMO-protein-intein(SPI)sandwich *** utility of SUMO enhances the protein fusion yield and solubility,prevents premature hydrolysis and simplifies the purification *** inaccessible protein thioester with internal Cys residues can be readily produced and is compatible with the EPL-desulfurization protocol used to prepare complex proteins,which is otherwise difficult to obtain using traditional *** utility has been highlighted through the synthesis of human granulocyte colony-stimulating factor(G-CSF).

关键词： Desulfurization Glycoproteins Glycopeptides protein expression Peptide thioester

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