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Construction of cholecystokinin transgenic mouse and its effects on food intake

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Chinese Medical Journal 2009年第17期122卷 2022-2026页

作者： LI Yong-ning LI Gui-lin ZHANG Lian-feng Department of Neurosurgery Peking Union Medical CollegeHospital Chinese Academy of Medical Science Beijing 100730China Key Laboratory of Human Comparative Medicine Institute ofLaboratory Animal Science Chinese Academy of MedicalSciences & Comparative Medical Center Peking Union MedicalCollege Beijing 100021 China

Background Cholecystokinin （CCK） is one of the richest neuropeptides in the mammalian brain, which is mainly distributed in the cerebral cortex, hippocampus, thalamus and caudate-putamen. CCK is implicated in a variety of behavioral functions such as food intake, learning, memory, anxiety, pain and neuroprotection. The current research results for CCK are obtained mainly through injection of CCK peptide into the body. The key issues of whether CCK can regulate diet by a central pathway and whether there are long-term regulation effects on diet are still unresolved. In this study, the effects of CCK on food intake in transgenic mice were investigated. Methods Transgenic mice were created by microinjection of the PDGF-CCK construct into male pronucleus of the zygotes. The genomic phonetype of transgenic mice were identified by PCR. The expression of PDGF-CCK was analyzed by Western blotting. Body weight, plasma glucose, cholesterol and triglycerides were assayed and analyzed. Results Two PDGF-CCK transgenic independent lines were established and exhibited a high-levels brain-specific transgene expression compared with that of nontransgenic littermate controls. The food intake of male CCK transgenic mice was decreased by 5%-10% with the same levels of water consumed compared with wild type mice. The food intake in female mice was not obviously changed. In the transgenic mice the bodyweight was lower and plasma glucose was higher compared with the nontransgenic littermate controls. Conclusions The high expression of the CCK gene in the brain can decrease body weight and increase plasma glucose. The differences in food intake between the males and females require further study. Chin Med J 2009; 122（17）：2022-2026

关键词： cholecystokinin transgene eating mouse

Histological analysis of human pancreatic carcinoma following irreversible electroporation in a nude mouse model

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World Journal of Gastrointestinal Oncology 2018年第12期10卷 476-486页

作者： Jun-Jun Su Kai Xu Peng-Fei Wang Hao-Yun Zhang Yong-Liang Chen Division of Gastroenterological Surgery Depart-ment of SurgeryShanxi Provincial People’s Hospital Department of Hepatobiliary Surgery Chinese People’s Liberation Army General Hospital

AIM To determine changes in the morphology and function of pancreatic cancer cells after irreversible electroporation(IRE) treatment, and to explore the clinical significance of IRE treatment for pancreatic cancer providing an experimental basis for the clinical application of IRE treatment. METHODS IRE was carried out in an athymic nude mouse model of pancreatic carcinoma generated with human pancreatic cancer cells 1. In therapy groups, IRE electrodes were inserted with 90 pulses per second at 800 V/cm applied to ablate the targeted tumor tissues. Histological assessment of the affected tissue was performed by hematoxylin and eosin staining(HE). Quantification of cell proliferation and apoptosis was performed by evaluating Ki67 and caspase-3 levels, respectively. Flow cytometry was used to assess cell apoptosis. Ultrasound imaging was carried out to evaluate IRE treatment results. Pathological correlation studies showed IRE is effective for the targeted ablation of pancreatic tumors in an orthotopic mouse model.RESULTS IRE was efficacious in removing tumors in the orthotopic mouse model. The IRE-ablated zone displays characteristics of nude mouse models at different time-points as assessed by hematoxylin and eosin staining. Immunohistochemical analysis of samples from the pancreatic cancer models showed significantly enhanced caspase-3 cleavage and Ki67. Flow cytometry data corroborated the above findings that apoptosis in tumor cells was observed immediately on the first postoperative day, and with time the middle and late stages of apoptosis were observed. For ultrasound imaging studies, the IRE ablation zone became a hyperechoic area due to increasing inflammatory and immunologic cellular contents. CONCLUSION IRE is a promising new approach for pancreatic cancer, with many potential advantages over conventional ablation techniques.

关键词： Irreversible electroporation Pancreatic carcinoma Pathological evaluation Transplantation model Nude mouse

Towards estimating computer users＇ mood from interaction behaviour with keyboard and mouse

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Frontiers of Computer Science 2013年第6期7卷 943-954页

作者： Iflikhar Ahmed KHAN Willem-Paul BRINKMAN Robert HIERONS Department of Computer Software Engineering University of Engineering & Technology Mardan 23200 Pakistan Interactive Intelligence Group Delft University of Technology Delft 2628B the Netherlands Department of Information Systems and Computing Brunel University West London UB83PH UK

The purpose of this exploratory research was to study the relationship between the mood of computer users and their use of keyboard and mouse to examine the possibility of creating a generic or individualized mood measure. To examine this, a field study （n = 26） and a controlled study （n = 16） were conducted. In the field study, interaction data and self-reported mood measurements were collected during normal PC use over several days. In the controlled study, participants worked on a programming task while listening to high or low arousing background music. Besides subjective mood measurement, galvanic skin response （GSR） data was also collected. Results found no generic relationship between the interaction data and the mood data. However, the re- suits of the studies found significant average correlations be- tween mood measurement and personalized regression models based on keyboard and mouse interaction data. Together the results suggest that individualized mood prediction is pos- sible from interaction behaviour with keyboard and mouse.

关键词： keyboard mouse interaction mood measure computer users programming

ENRICHMENT OF THE SPINDLE FIBER ATTACHMENTS FROM mouse LIVER CELL NUCLEI

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Science China Chemistry 1989年第7期32卷 816-821页

作者：施履吉倪祖梅陆仕芳王阁祁慧霖苏庆祥 Shanghai Institute of Cell Biology Academia Sinica

Spindle fiber attachments (SFAs) were enriched in a fraction of nuclei isolated from mouselivers. The enrichment method combines sonication, treatment with 2 mol/L NaCl and highspeed centrifugation. SFA was enriched 27-fold on the average (n = 4) when measured byradioimmunoassay. The basic method offers opportunities for further increases of yield andfor the enrichment of SFA uf other vertebrates.

关键词： spindle fiber attachment enrichment isolated liver nuclei mouse

Current humanized mouse models for studying human immunology and HIV-1 immuno-pathogenesis

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Science China(Life Sciences) 2010年第2期53卷 195-203页

作者： MEISSNER Eric Lineberger Comprehensive Cancer Center Department of Microbiology and ImmunologySchool of MedicineUniversity of North Carolina at Chapel Hill

A robust animal model for "hypothesis-testing/mechanistic" research in human immunology and immuno-pathology should meet the following criteria.First,it has well-studied hemato-lymphoid organs and target cells similar to those of humans.Second,the human pathogens establish infection and lead to relevant diseases.Third,it is genetically inbred and can be manipulated via genetic,immunological and pharmacological means.Many human-tropic pathogens such as HIV-1 fail to infect murine cells due to the blocks at multiple steps of their life cycle.The mouse with a reconstituted human immune system and other human target organs is a good candidate.A number of human-mouse chimeric models with human immune cells have been developed in the past 20 years,but most with only limited success due to the selective engraftment of xeno-reactive human T cells in hu-PBL-SCID mice or the lack of significant human immune responses in the SCID-hu Thy/Liv mouse.This review summarizes the current understanding of HIV-1 immuno-pathogenesis in human patients and in SIV-infected primate models.It also reviews the recent progress in the development of humanized mouse models with a functional human immune system,especially the recent progress in the immunodeficient mice that carry a defective gammaC gene.NOD/SCID/gammaC-/(NOG or NSG) or the Rag2-/-/gammaC-/double knockout (DKO) mice,which lack NK as well as T and B cells (NTB-null mice),have been used to reconstitute a functional human immune system in central and peripheral lymphoid organs with human CD34+ HSC.These NTB-hu HSC humanized models have been used to investigate HIV-1 infection,immuno-pathogenesis and therapeutic interventions.Such models,with further improvements,will contribute to study human immunology,human-tropic pathogens as well as human stem cell biology in the tissue development and function in vivo.

关键词： Humanized mouse models HIV-1 NOD/SCID/gammaC-/- Rag2-/-/gammaC-/-

INT-767 improves histopathological features in a dietinduced ob/ob mouse model of biopsy-confirmed nonalcoholic steatohepatitis

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World Journal of Gastroenterology 2018年第2期24卷 195-210页

作者： Jonathan D Roth Michael Feigh Sanne S Veidal Louise KD Fensholdt Kristoffer T Rigbolt Henrik H Hansen Li C Chen Mathieu Petitjean Weslyn Friley Niels Vrang Jacob Jelsing Mark Young Intercept Pharmaceuticals Intercept PharmaceuticalsSan DiegoCA 92121United States Gubra Hoersholm DK-2970Denmark PharmaNest Genesis Imaging ServicesPrincetonNJ 08540United States Qualyst Transporter Solutions DurhamNC 27713United States

AIM To characterize the efficacy of the dual FXR/TGR5 receptor agonist INT-767 upon histological endpoints in a rodent model of diet-induced and biopsy-confirmed non-alcoholic steatohepatitis(NASH).METHODS The effects of INT-767 on histological features of NASH were assessed in two studies using Lep^(ob/ob)(ob/ob) NASH mice fed the AMLN diet(high fat with transfat, cholesterol and fructose). In a proof-of-conceptstudy, Lep^(ob/ob)(ob/ob) NASH mice were first dosed with INT-767(3 or 10 mg/kg for 8 wk). A second ob/ob NASH study compared INT-767(3 and 10 mg/kg) to obeticholic acid(OCA)(10 or 30 mg/kg; 16 wk). Primary histological endpoints included qualitative and quantitative assessments of NASH. Other metabolic and plasma endpoints were also assessed. A comparative assessment of INT-767 and OCA effects on drug distribution and hepatic gene expression was performed in C57 Bl/6 mice on standard chow. C57 Bl/6 mice were orally dosed with INT-767 or OCA(1-30 mg/kg) for 2 wk, and expression levels of candidate genes were assessed by RNA sequencing and tissue drug levels were measured by liquid chromatography tandem-mass spectrometry.RESULTS INT-767 dose-dependently(3 and 10 mg/kg, PO, QD, 8 wk) improved qualitative morphometric scores on steatohepatitis severity, inflammatory infiltrates and fibrosis stage. Quantitative morphometric analyses revealed that INT-767 reduced parenchymal collagen area, collagen fiber density, inflammation(assessed by Galectin-3 immunohistochemistry) and hepatocyte lipid droplet area following INT-767 treatment. In a comparative study(16 wk), the FXR agonists OCA(10 and 30 mg/kg) and INT-767(3 and 10 mg/kg) both improved NASH histopathology, with INT-767 exerting greater therapeutic potency and efficacy than OCA. Mechanistic studies suggest that both drugs accumulate similarly within the liver and ileum, however, the effects of INT-767 may be driven by enhanced hepatic, but not ileal, FXR function. CONCLUSION These findings confirm the potential utility of FXR and dual FXR/TGR5 activation a

关键词： Non-alcoholic steatohepatitis INT-767 Obeticholic acid Liver biopsy FXR TGR5 mouse model

Platelet-rich plasma enhances adipose-derived stem cell-mediated angiogenesis in a mouse ischemic hindlimb model

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World Journal of Stem Cells 2018年第12期10卷 212-227页

作者： Chia-Fang Chen Han-Tsung Liao Department of Plastic and Reconstructive Surgery Chang Gung Memorial Hospital Craniofacial Research Center Chang Gung Memorial Hospital College of Medicine Chang Gung University

AIM To evaluate the angiogenic effect of platelet-rich plasma(PRP)-preconditioned adipose-derived stem cells(ADSCs) both in vitro and in a mouse ischemic hindlimb model.METHODS ADSCs were divided based on culture medium: 2.5% PRP, 5% PRP, 7.5% PRP, and 10% PRP. Cell proliferation rate was analyzed using the MTS assay. The gene expression of CD31, vascular endothelial growth factor, hypoxia-inducible factors, and endothelial cell nitric oxide synthase was analyzed using reverse transcription polymerase chain reaction. Cell markers and structural changes were assessed through immunofluorescence staining and the tube formation assay. Subsequently, we studied the in vivo angiogenic capabilities of ADSCs by a mouse ischemic hindlimb model.RESULTS The proliferation rate of ADSCs was higher in the 2.5%, 5%, and 7.5% PRP groups. The expression of hypoxia-inducible factor, CD31, vascular endothelial growth factor, and endothelial cell nitric oxide synthase in the 5% and 7.5% PRP groups increased. The 5%, 7.5%, and 10% PRP groups showed higher abilities to promote both CD31 and vascular endothelial growth factor production and tubular structure formation in ADSCs. According to laser Doppler perfusion scan, the perfusion ratios of ischemic limb to normal limb were significantly higher in 5% PRP, 7.5% PRP, and human umbilical vein endothelial cells groups compared with the negative control and fetal bovine serum(FBS) groups(0.88 ± 0.08, 0.85 ± 0.07 and 0.81 ± 0.06 for 5%, 7.5% PRP and human umbilical vein endothelial cells compared with 0.42 ± 0.17 and 0.54 ± 0.14 for the negative control and FBS, P < 0.01).CONCLUSION PRP-preconditioned ADSCs presented endothelial cell characteristics in vitro and significantly improved neovascularization in ischemic hindlimbs. The optimal angiogenic effect occurred in 5% PRP-and 7.5% PRPpreconditioned ADSCs.

关键词： Platelet-rich plasma Adipose-derived stem cells Mesenchymal stem cell Angiogenesis Endothelial differentiation mouse ischemic hindlimb model

A Study of Protein and Glycoprotein Syntheses in Pre-implantation mouse Embryos Using Mini-2D-Electrophoresis, Video Densitometer Scanning and Computer-Image Analysis

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Science China Chemistry 1993年第1期36卷 39-44页

作者：郑明曹詠清 B.LoftsDepartment of Zoology University of Hong Kong Hong Kong 陈廷汉 Department of Endocrinology and Laboratory of Reproductive Biology Institute of Zoology Academia Sinica Beijing 100080 PRC Department of Zoology University of Hong Kong Hong Kong

In the present study, proteins and glycoproteins of mouse embryos at 2-cell,morula and blastocyst stages were analyzed.The techniques of 35S-Met incorporation,ConA antiserum-precipitating ConA-binding proteins, mini-2D-electrophoresis, fluorography,video densitometer scanning and the computer-lmage system were used for analyses.Results of the investigations indicated that proteins and glycoproteins were synthesized by the embryos in a gradual increase manner from 2-cell to blastocyst. A relatively large amount of glycoproteins was synthesized during the morula and blastocyst stages.Two specific small glycoproteins respectively with molecular weights about 6500 and 9000 and PIs at 5.0 and 6.5 were apparently synthesized in the blastocyst but not in the 2-cell or the morula.

关键词： implantation mouse embryos glyeoproteins mini 2D-electrophoresis computer-image analysis.

A quantification model for apoptosis in mouse embryos in the early stage of fetation

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Science China(Life Sciences) 2009年第10期52卷 922-927页

作者： WANG PengFei1, FU JianHua1, MA WanYun1, CHEN DieYan1, Lü DanYu2 & BAI WenJia3 1 Key Laboratory for Atomic and Molecular Nanosciences of Education Ministry, Department of Physics, Tsinghua University, Beijing 100084, China 2 Department of Anatomy & Histology and Embryology, School of Basic Medical Sciences Peking University Health Science Center, Beijing 100083, China 3 Department of Automation, Tsinghua University, Beijing 100084, China Key Laboratory for Atomic and Molecular Nanosciences of Education Ministry Department of Physics Tsinghua University Beijing China Department of Anatomy & Histology and Embryology School of Basic Medical Sciences Peking University Health Science Center Beijing China Department of Automation Tsinghua University Beijing China

Apoptosis is the most important inducement and modulator for embryos in the early stage of fetation, i.e. after the 8-cell stage, mostly the morula and blastula stage, to proceed to the stage of nonlinear development. Using a two-photon laser scanning microscopy (TPLSM) system, we obtained 3-dimensional (3D) fluorescent images of preimplantation mouse embryos. A model for quantification was established. The statistical results for the spatial location of apoptosis bodies in embryos was obtained following image processing, as well as investigation of the kinetics of apoptosis. It was found that most (70%) apoptosis occurred in the trophectoderm, and the departure between the centroid and geometric center of embryos had a step transition when embryos developed into the 32-cell stage, which was consistent with the theoretical prediction that the blastocele would induce a symmetry break of the distribution of cells in embryos.

关键词： apoptosis preimplantation mouse embryo TPLSM quantification model