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Occurrence of cGMP/nitric oxide-sensitive store-operated calcium entry in fibroblasts and its effect on matrix metalloproteinase secretion

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World Journal of Gastroenterology 2006年第34期12卷 5483-5489页

作者： Yong huang Min-Qiang Lu hua li Chi xu Shu-Hong Yi Gui-hua Chen liver Transplantation Centre the Third Affiliated Hospital Sun Yat-Sen University Guangzhou 510630 Guangdong Province China

AIM： To examine the existence of Nitric oxide/ cGMP sensitive store-operated Ca^2＋ entry in mouse fibroblast NIH/3T3 cells and its influence on matrix metalloproteinase （MMP） production and adhesion ability of fibroblasts. METHODS： NIH/3T3 cells were cultured. Confocal laser scanning microscopy was used to examine the existence of thapsigargin-induced store-operated Ca^2＋ entry in fibroblasts. Gelatin zymography and semiquantitative reverse transcriptase-polymerase chain reaction （RTPCR） were employed to detect the involvement of [Ca^2＋]i and NO/cGMP in MMP secretion. The involvement of NO/ cGMP-sensitive Ca^2＋ entry in adhesion was determined using matrigel-coated culture plates. RESULTS： 8-bromo-cGMP inhibited the thapsigargin-induced Ca^2＋ entry in 3T3 cells. The cGMP-induced inhibition was abolished by an inhibitor of protein kinase G, KT5823 （1μmol/L）. A similar effect on the Ca^2＋ entry was observed in 3T3 cells in response to a NO donor, （±）-S-nitroso-N-acetylpenicillamine （SNAP）. The inhibitory effect of SNAP on the thapsigargin-induced Ca^2＋ entry was also observed, indicating NO/cGMP-regulated Ca^2＋ entry in 3T3 cells. Results of gelatin zymography assay showed that addition of extracellular Ca^2＋ concentration induced MMP release and activation in a dose-dependent manner. RT-PCR also showed that cGMP and SNAP reduced the production of MMP mRNA in 3T3 cells. Experiments investigating adhesion potentials demonstrated that cGMP and SNAP could upgrade 3T3 cell attachment rate to the matrigel-coated culture plates.CONCLUSION： NO/cGMP sensitive store-operated Ca^2＋ entry occurs in fibroblasts, and attenuates their adhesion potentials through its influence on MMP secretion.

关键词： cGMP Nitric oxide Protein kinase G Storeoperated Ca^2+ entry Matrix metalloproteinase Fibroblast

Characterization of Multi-layer Weld Metal and Creep–Rupture Behavior of Modified 10Cr–1Mo Welded Joint

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Acta Metallurgica Sinica(English Letters) 2020年第6期33卷 808-820页

作者： Chang-Zhen Zhang Chen-Dong Shao Hai-Chao Cui hua-li xu Feng-Gui Lu Shanghai Key Laboratory of Materials Laser Processing and Modification School of Materials Science and EngineeringShanghai Jiao Tong UniversityShanghai 200240China

The rupture behavior of the modified 10Cr–1Mo steel multi-layer welded joint is determined by the fine-grain zones of the weld metal adjacent to the fusion line during the long-term creep test at 620℃. The microstructures of multi-layer weld metal before and after the creep tests were characterized in detail, and its role in creep behavior was systematically investigated. Most grain boundaries of subgrains represented the low-angle boundaries in the weld metal adjacent to the fusion line both before and after the creep test. The widths of grains in the fine-grain zones were about 0.5–1 μm. The fracture morphology appeared as "wave" structure due to the cracking initiating from multi-layer grain boundaries in the fine-grain zones. Some W elements that melted into weld metal adjacent to the fusion line altered the thermodynamic and kinetic conditions of the Laves phase formation during long-term creep exposure. Laves phase particles mainly distributed along the grain boundaries due to the faster diffusion and segregation of Mo, W, and Si elements. Moreover, higher-density grain boundaries in the fine-grain zones led to easier nucleation and growth of Laves phase particles. Compared with other areas in the welded joint, the size of Laves phase particles in the fine-grain zones of the weld metal adjacent to the fusion line was the largest ones. The interface between Laves phase particles and the matrix acted as the nucleation site of creep micro-cavities. The creep micro-cavities grew up at the expense of fine-grain boundaries and even grew across the grain boundary deeply into adjacent grains, and then developed to cracks in the fine-grain zones.

关键词： 10Cr–1Mo Welded joint Fine grains Multi-layer Creep rupture

p53-expressing conditionally replicative adenovirus CNHK500-p53 against hepatocellular carcinoma in vitro

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World Journal of Gastroenterology 2007年第5期13卷 683-691页

作者： Hong-Chuan Zhao Qi Zhang Yang Yang Min-Qiang Lu hua li Chi xu Gui-hua Chen liver Transplantation Centre the Third Affiliated Hospital Sun Yat-Sen University Guangzhou 510630 Guangdong Province China

AIM： To develop a conditionally replicative gene-viral vector system called CNHK500-p53, which contains dual promoters within the E1 region, and combines the advantages of oncolytic virus and gene therapies for hepatocellular carcinoma （HCC）. METHODS： CNHK500-p53 was constructed by using human telomerase reverse transcriptase （hTERT） promoter to drive adenovirus E1a gene and hypoxia response element （HRE） promoter to drive adenovirus E1b gene. p53 gene expressing cassette was inserted into the genome of replicative virus. Viral replication experiments, cytopathic effect （CPE） and methyl thiazolyl tetrazolium （MTT） assay were performed to test the selective replication and oncolytic efficacy of CNHK500-p53. RESULTS： Immunohistochemistry verified that infection with CNHK500-p53 was associated with selective replication of adenovirus and production of p53 protein in telomerase-positive and hypoxia-inducible factordependent HCC cells, p53 protein secreted from HepG2, infected with CNHK500-p53 was significantly higher than that infected with nonreplicative adenovirus Ad-p53 in vitro （388 ± 34.6 μg/L vs 76.3 ± 13.17 μg/L）. Viral replication experiments showed that replication of CNHK500-p53 and CNHK500 or WtAd5, was much stronger than that of Ad-p53 in tested HCC cell lines. CPE and H1-F assay indicated that CNHK500-p53 selectively replicated in and killed HCC cells while leaving normal cells unaffected. CONCLUSION： A more efficient gene-viral system is developed by combining selective oncolysis with exogenous expression of p53 against HCC cells.

关键词： Conditionally replicative adenovirus Oncolytic virotherapy Gene therapy p53 gene Hepatocellular carcinoma

Microglial depletion impairs glial scar formation and aggravates inflammation partly by inhibiting STAT3 phosphorylation in astrocytes after spinal cord injury

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Neural Regeneration Research 2023年第6期18卷 1325-1331页

作者： Zhi-Lai Zhou huan Xie Xiao-Bo Tian hua-li xu Wei li Shun Yao Hui Zhang The Spine Surgery Department Guangdong Second Provincial General HospitalGuangzhouGuangdong ProvinceChina The Second School of Clinical Medicine Southern Medical UniversityGuangzhouGuangdong ProvinceChina Department of Anesthesiology ZhuJiang HospitalSouthern Medical UniversityGuangzhouGuangdong ProvinceChina

Astrocytes and microglia play an orchestrated role following spinal cord injury;however,the molecular mechanisms through which microglia regulate astrocytes after spinal cord injury are not yet fully ***,microglia were pharmacologically depleted and the effects on the astrocytic response were *** further explored the potential mechanisms involving the signal transducers and activators of transcription 3(STAT3)*** in vivo experiments,we constructed a contusion spinal cord injury model in C57BL/6 *** deplete microglia,all mice were treated with colony-stimulating factor 1 receptor inhibitor PLX3397,starting 2 weeks prior to surgery until they were *** proliferation was examined by 5-ethynyl-2-deoxyuridine(EdU)and three pivotal inflammatory cytokines were detected by a specific Bio-Plex Pro^(TM) Reagent *** function,neuroinflammation,astrocyte activation and phosphorylated STAT3(pSTAT3,a maker of activation of STAT3 signaling)levels were *** in vitro experiments,a microglia and astrocyte coculture system was established,and the small molecule STA21,which blocks STAT3 activation,was applied to investigate whether STAT3 signaling is involved in mediating astrocyte proliferation induced by ***3397 administration disrupted glial scar formation,increased inflammatory spillover,induced diffuse tissue damage and impaired functional recovery after spinal cord *** depletion markedly reduced EdU+proliferating cells,especially proliferating astrocytes at 7 days after spinal cord *** sequencing analysis showed that the JAK/STAT3 pathway was downregulated in mice treated with *** immunofluorescence staining confirmed that PLX3397 significantly decreased STAT3 expression in ***,in vitro coculture of astrocytes and microglia showed that microglia-induced astrocyte proliferation was abolished by STA21 *** findings suggest that microglial depl

关键词： astrocytes coculture colony-stimulating factor 1 receptor inhibitor EdU glia scar inflammatory response microglia phosphorylation proliferation spinal cord injury STAT3

Smith Prediction Monitor AGC System Based on Fuzzy Self-Tuning PID Control

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Journal of Iron and Steel Research International 2010年第2期17卷 22-26页

作者： SUN Jie ZHANG Dian-hua li xu ZHANG Jin DU De-shun State Key Laboratory of Rolling and Automation Northeastern University Shenyang 110004 Liaoning China

In accordance with the feature of pure delay in monitor AGC system for cold rolling mill, a new fuzzy selftuning PID Smith prediction controller is developed. The position control model is deduced based on a single stand cold rolling mill, and the fuzzy controller for monitor AGC system is designed. The analysis of dynamic performance for traditional PID Smith prediction controller and fuzzy self-tuning PID Smith prediction controller is done by MAT- LAB toolbox. The simulation results show that fuzzy self-tuning PID Smith controller has stronger robustness, faster response and higher static accuracy than traditional PID Smith controller.

关键词： monitor AGC fuzzy self-tuning PID control Smith predictor position control

Characteristics of global cerebral ischemia models constructed by modified four-vessel occlusion in rats

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Neural Regeneration Research 2006年第5期1卷 436-440页

作者： Jinbao li Lai Jiang hua xu Yuanchang Xiong Xiaoming Deng Department of Anesthesiology Changhai Hospital Second Military Medical University of ChinesePLA Shanghai 200433 China

BACKGROUND： Pulsinelli et al developed a kind of rat models with four-vessel occlusion-induced global cerebral ischemia, Because the histo-pathological changes and severe cerebral ischemia reproducibility of this model are good and the stability of this model in circulation respiration is superior to that of other models, so four-vessel occlusion method has become a classic modeling method for global cerebral ischemia model, This model has been improved in some laboratories to meet different requirements in different studies. OBJECTIVE： To establish a highly reproducible rat model of reversible forebrain ischemia by modifying four-vessel occlusion model introduced by Pulsinelli et al, and to investigate its neurophysiological and pathological changes and the characteristics of modified operation. DESIGN ： Completely randomized grouping, controlled tria. SETTING ： Department of Anesthesiology,Changhai Hospital, Second Military Medical University of Chinese PLA. MATERIALS ： A total of 65 male healthy SD rats, weighing 250-300 g, were provided by the Experimental Animal Center of Second Military Medical University of Chinese PLA. VSM hemodynamic monitor and temperature monitor （Thermal ert TH-5, U.S.A） were used. METHODS ： The trial was conducted in the Department of Anesthesiology, Changhai Hospital, Second Military Medical University of Chinese PLA from January 2005 to March 2006. ① Experimental grouping： Sixty-five rats were randomly divided into the following 7 groups： sham-operation group （n =9）： given the same operation, without occlusion of vessels; 5 minutes ischemia group （n =9）： ischemia 5 minutes and reperfusion 72 hours; 10 minutes ischemia group （n =8）： ischemia 10 minutes and reperfusion 72 hours; 15 minutes ischemia group （n =9）： ischemia 15 minutes and reperfusion 72 hours; 20 minutes ischemia group （n =8）： ischemia 20 minutes and reperfusion 72 hours; 30 minutes ischemia group （n =7）; ischemia 30 minutes and reperfusion 72 hours; ischemia con

关键词： cerebral ischemia blood pressure rectal temperature arterial blood gas dentate gyrus common carotid artery pathological change Experimental Animal rat model carotid arteries result analysis modeling method blood flow

Overexpression of eRF3a Promotes Cell Proliferation and Migration in liver Cancer

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Current Medical Science 2022年第1期42卷 100-107页

作者： Yi-qing XI li-hua xu li-jie YANG hua-qiao WANG Tie-cheng YANG Zhi li Wei XIE Jing-wei ZHANG xuan-fei li Mao-hui FENG Department of Gastrointestinal Surgery Zhongnan Hospital of Wuhan UniversityWuhan 430071China Clinical Medical Research Center of Peritoneal Cancer of Wuhan 430071 China Clinical Cancer Study Center of Hubei Province Wuhan 430071China Hubei Key Laboratory of Tumor Biological Behaviors Wuhan 430071China

Objective:The eukaryotic release factor 3a(eRF3a),a member of the eukaryotic peptide chain release factor family,is overexpressed in several types of *** study aims to investigate the biological role and mechanism of ... 详细信息

关键词： eukaryotic release factor 3a liver cancer migration invasion

GSPT1 Functions as a Tumor Promoter in Human liver Cancer

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Current Medical Science 2023年第1期43卷 104-114页

作者： Yi-qing XI Jing-bo GAO xuan-fei li li-hua xu Zhi-li li-jie YANG Jing WANG hua-qiao WANG Xiao-chang FANG Si-rui huaNG Wei XIE Mao-hui FENG Jing-wei ZHANG Department of Breast and Thyroid Surgery Zhongnan Hospital of Wuhan UniversityWuhan430071China Department of Gastrointestinal Surgery Zhongnan Hospital of Wuhan UniversityWuhan430071China Hubei Cancer Clinical Study Center Wuhan UniversityWuhan430071China Institute of Hydrobiology Chinese Academy of SciencesWuhan430071China

Objective This study analyzed the role of G1 to S phase transition 1 protein(GSPT1)in promoting progression of liver cancer *** A bioinformatics database was used to analyze the expression levels of GSPT1 in liver cancer tissues and the prognosis of ***,Western blotting and quantitative PCR were used to verify the expression levels of GSPT1 between normal hepatocytes and hepatoma *** used a CRISPR/Cas9 system to construct knockouts of GSPT1 in HepG2 and HCCLM9 liver cancer *** effect of GSPT1 on liver cancer cell migration and invasion was analyzed using flow cytometry,migration,and tumor formation *** The Cancer Genome Atlas liver Hepatocellular Carcinoma dataset indicated that GSPT1 expression was upregulated in liver cancer cell lines,and patients with liver cancer had poor *** of GSPT1 in cells significantly inhibited tumor proliferation,cell migration,and growth in *** In this study,we found that GSPT1 promotes the migration and invasion of liver cancer cells.

关键词： Gl to S phase transition I protein liver cancer migration invasion

Establishment and characterization of a novel nasopharyngeal carcinoma cell line (SUNE2) from a Cantonese patient

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Chinese Journal of Cancer 2012年第1期31卷 36-44页

作者： Ju-Qin Dong Man-Zhi li Zhi-Gang liu Qian Zhong Dan Xiong li-hua xu Yong Du Yun-Fei Xia Mu-Sheng Zeng State Key Laboratory of Oncology in South China GuangzhouGuangdong 510060P.R.China Department of Experimental Research Department of Radiation Therapy Sun Yat-sen University Cancer CenterGuangzhouGuangdong 510060P.R.China.

The undifferentiated form of nasopharyngeal carcinoma (NPC) is the most common malignant head and neck cancer in South China, especially in Cantonese populations. However, few NPC cell lines have been established from the patients in this region. In this study, we established a new NPC cell line, termed SUNE2, from a Cantonese patient with undifferentiated NPC. This cell line had extremely low concentrations of Epstein-Barr virus (EBV) DNA in long-term culture and expressed low levels of latent membrane protein 1 (LMP1), latent membrane protein 2A (LMP2A), BamH1-A right frame 1 (BARF1), EBV-encoded RNA-1 (EBER1), and EBV-encoded RNA-2 (EBER2) in early passages. SUNE2 cells also showed much stronger transforming ability than 5-8F cells in colony formation assays and anchorage- independent growth assays in soft agar, and they only need 2 weeks to form tumors in nude mice. In summary, the SUNE2 cell line is a new in vitro model that can be used for further research on the mechanisms underlying the occurrence and development of NPC.

关键词：细胞株鼻咽癌潜伏膜蛋白1 粤语表征 EB病毒恶性肿瘤生长实验