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Quantifying E2F1 protein dynamics in single cells

Quantitative Biology 2020年第1期8卷 20-30页

作者： Bernard Mathey-Prevot Bao-Tran Parker Carolyn Im Cierra Hong Peng Dong Guang Yao Lingchong You department of pharmacology and Cancer Biology Duke University School of MedicineDurhamNC 27710USA Duke University School of Medicine DurhamNC 27710USA molecular and Cellular Biology University of ArizonaTucsonAZ 85721USA department of Biomedical Engineering Duke UniversityDurhamNC 27705USA Center for Genomic and Computational Biology Duke UniversityDurhamNC 27705USA department of molecular genetics and Microbiology Duke University School of MedicineDurhamNC 27708USA department of Pediatrics Duke University School of MedicineDurhamNC 27708USA University of North Carolina School of Medicine Chapel HillNC 27516USA Janelia Research Campus Howard Hughes Medical InstituteAshburnVA 20147USA

Background:E2F1 protein,a major effector of the Rb/E2F pathway plays a central role in regulating cell-fate decisions involved in proliferation,apoptosis,and *** expression is highly dynamic and tightly modulated through a combination of transcriptional,translational and posttranslational ***,the mechanisms by which its expression and activity can promote different cellular outcomes remain to be fully *** better document E2F1 expression in live cells,we have engineered a series of fluorescent E2F1 protein reporters that quantitatively capture E2F1 protein ***:Reporter constructs,under the control of the mouse or human E2F1 proximal promoter,were designed to express an E2F1-Venus fusion protein incapable of binding *** addition,constructs either included or excluded the y untranslated region(3fUTR)of the E2F1 *** constructs were introduced into fibroblasts and epithelial cells,and expression of the fusion reporter protein was validated and quantified in single cells using live ***:In all cases,expression of the reporter protein effectively recapitulated the behavior of E2F1 under various conditions,including cell cycle progression and genotoxic *** or little fluorescent signal of the reporter was detected in G〇,but as the cycle progressed,expression of the reporter protein steadily increased in the nucleus,peaking a few hours before cell division,but declining to baseline 2-3 h prior to the onset of *** absence of the E2F13 fUTR in the constructs led to considerably higher steady-state levels of the fusion protein,which although normally regulated,exhibited a slightly less complex dynamic profile during the cell cycle or genotoxic ***,the presence or absence of Rb failed to impact the overall detection and levels of the reporter ***:Our validated E2F1 protein reporters complement nicely other reporters of the Rb/E2F pathway and provide a unique tool to f

关键词： protein E2F1 reporter cell cycle

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Complete or partial trisomy 3 in gastro-intestinal MALT lymphomas co-occurs with aberrations at 18q21 and correlates with advanced disease stage:A study on 25 cases

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World Journal of Gastroenterology 2005年第46期11卷 7384-7385页

作者： Jens Krugmann Alexandar Tzankov Stephan Dirnhofer Falko Fend Dominik Wolf Reiner Siebert Pensiri Probst Martin Erdel Institute of PathologyMedical University of Innsbruck Austria Department of Medical GeneticsMolecular and Clinical PharmacologyMedical University of InnsbruckAustria Institute of PathologyMedical University of Innsbruck Austria Institute of PathologyUniversity of Basel Switzerland Institute of PathologyTechnical University of Munich Germany department of Internal MedicineDivision of Hematology/Oncology Medical University of InnsbruckAustria Institute of Human geneticsUniversity Hospital Schleswig-Holstein Campus Kiel Germany department of Medical geneticsmolecular and Clinical pharmacology Medical University of InnsbruckAustria

Taji et al . have reported in their study on 13 patients with gastric mucosa-associated lymphoid tissue （MALT） lymphomas an aggressive tumor course in trisomy 3 positive cases. The authors analyzed only stage I patients with classical low-grade marginal zone lymphoma of the MALT type and detected the trisomy 3 using an alphasatellite DNA probe directed to the centromere. Their data support the observation that trisomy 3 is the most frequent cytogenetic aberration in MALT lymphomas .

关键词： MALT lymphoma Trisomy 3 Trisomy 18

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A complex signature network that controls the upregulation of PRMT5 in colorectal cancer

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Genes & Diseases 2022年第2期9卷 285-287页

作者： Han Wei Antja-Voy Hartley Aishat Motolani Guanglong Jiang Ahmad Safa Lakshmi Prabhu Yunlong Liu Tao Lu department of pharmacology and Toxicology Indiana University School of Medicine635 Barnhill DriveIndianapolisIN 46202USA department of Medical&molecular genetics Indiana University School of Medicine975 W.Walnut StreetIndianapolisIN 46202USA department of Biochemistry and molecular Biology Indiana University School of Medicine635 Barnhill DriveIndianapolisIN 46202USA Current address:Beckman Coulter Life Sciences 5350 Lake-view Pkwy S DrIndianapolisIN 46268USA Current address:Dana-Farber Cancer Institute Harvard Med-ical School450 Brookline Ave.BostonMA 02215USA Current address:Gilead Sciences Inc.333 Lakeside Drive Foster CityCA 94404USA

Protein arginine methyltransferase 5(PRMT5)deregulation has emerged as an important prognostic indicator in human *** aberrant methylation-mediated epige-netic modification of signaling molecules,PRMT5 over-expression contributes to dysregulation of a variety of cellular processes related to cancer progression.1 However,the mechanisms governing PRMT5 expression levels in cancer remain largely *** this study,we examined factors that regulate PRMT5 expression at multiple levels.

关键词： cancer PRMT5 colorectal

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Synchronous multiple primary gastrointestinal cancers with CDH1 mutations:A case report

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World Journal of Clinical Cases 2019年第13期7卷 1703-1710页

作者： Mu-Ni Hu Wei Lv Rui-Yue Hu Yi-Fan Si Xiao-Wen Lu Yan-Juan Deng Huan Deng Medical College Nanchang University Nanchang 330006 Jiangxi Province China department of pharmacology Jiangxi Provincial People’s Hospital Nanchang 330006 Jiangxi Province China department of Gastroenterology the Fourth Affiliated Hospital of Nanchang University Nanchang 330003 Jiangxi Province China department of Radiology the Fourth Affiliated Hospital of Nanchang University Nanchang 330003 Jiangxi Province China department of Pathology the Fourth Affiliated Hospital of Nanchang University Nanchang 330003 Jiangxi Province China molecular Medicine and genetics Center the Fourth Affiliated Hospital of Nanchang University Nanchang 330003 Jiangxi Province China Renmin Institute of Forensic Medicine in Jiangxi Nanchang 330006 Jiangxi Province China

BACKGROUND Synchronous multiple primary cancers (SMPC) mean two or more malignant tumors occurring simultaneously and with different origins no matter what types they are or where they are located. The carcinogenesis of SMPC often involves variations of some specific genes. However, the correlation between CDH1 mutations and synchronous multiple primary gastrointestinal cancers is largely unknown. CASE SUMMARY A 62-year-old woman had sustained abdominal pain for one week and visited our hospital. Gastrointestinal endoscopy revealed multiple small polypoid lesions in both the stomach and colorectum. Computed tomography and laboratory results were within normal limits. Pathological evaluation confirmed signet ring cell carcinoma without obvious metastatic evidence. Malignant cells showed negativity for E-cadherin and positivity for β-catenin in the cytoplasm and nucleus. DNA sequencing performed on paraffin-embedded tissue revealed two exactly coincident alterations in CDH1, C.57T>G and C.1418A>T. CONCLUSION This case suggests that the combination of CDH1 mutations and WNT/β-catenin signaling activation contributes to the carcinogenesis of gastrointestinal SMPC.

关键词： Multiple primary cancers CDH1 Signet ring cell carcinoma Stomach Colon Case report

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In vitro laboratory infection research in orthopaedics:Why,when,and how

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World Journal of Orthopedics 2023年第8期14卷 598-603页

作者： Konstantinos Tsikopoulos Lorenzo Drago Gabriele Meroni Dimitrios Kitridis Byron Chalidis Fotios Papageorgiou Paraskevi Papaioannidou 1^(st)department of pharmacology School of MedicineFaculty of Health SciencesAristotle University of ThessalonikiThessaloniki 54124Greece department of Biomedical Sciences for Health School of MedicineUniversity of MilanMilan 20133Italy department of Biomedical Surgical and Dental SciencesUniversity of MilanMilan 20133Italy 1^(st)department of Orthopaedic School of MedicineFaculty of Health SciencesAristotle University of ThessalonikiThessaloniki 57010Greece department of Orthopaedic Surgery 404 General Army HospitalLarisa 41222Greece 1^(st)department of pharmacology Aristotle University of ThessalonikiThessaloniki 54124Greece

The musculoskeletal system involves multiple tissues which are constantly exposed to being exposed to various biological and mechanical *** such,isolating and studying a particular system from a complex human clinical environment is not always a realistic *** top of that,recruitment limitations,in addition to the nature of orthopaedic interventions and their associated cost,sometimes preclude consideration of human trials to answer a clinical ***,in this mini review,we sought to rationalize the rapid evolution of biomedical research at a basic scientific level and explain why the perception of orthopaedic conditions has fundamentally changed over the last *** more detail,we highlight that the number of orthopaedic in vitro publications has soared since *** but not least,we elaborated on the minimum requirements for conducting a scientifically sound infection-related laboratory experiment to offer valuable information to clinical *** also explained the rationale behind implementing molecular biology techniques,ex vivo experiments,and artificial intelligence in this type of laboratory research.

关键词： Biofilm In vitro Laboratory research Methodology

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Systemic epigallocatechin gallate protects against retinal degeneration and hepatic oxidative stress in the P23H-1 rat

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Neural Regeneration Research 2022年第3期17卷 625-631页

作者： Lorena Perdices Lorena Fuentes-Broto Francisco Segura Ana Cavero Elvira Orduna-Hospital Gema Insa-Sánchez Ana Isabel Sánchez-Cano Laura Fernández-Sánchez Nicolás Cuenca Isabel Pinilla Aragón Institute for Health Research(IIS Aragón) ZaragozaSpain pharmacology Physiology&Legal and Forensic MedicineUniversity of ZaragozaZaragozaSpain department of Applied Physics University of ZaragozaZaragozaSpain Health Sciences Faculty San Jorge UniversityVillanueva de GállegoSpain department of Ophthalmology Miguel Servet University HospitalZaragozaSpain department of Physiology Genetics and MicrobiologyUniversity of AlicanteAlicanteSpain department of Surgery Gynecology and ObstetricsUniversity of ZaragozaZaragozaSpain/Department of OphthalmologyLozano Blesa University HospitalZaragozaSpain

Retinitis pigmentosa(RP) is a group of inherited retinal disorders that lead to photoreceptor *** has been reported to be related to oxidative stress,autophagy,and inflammation.(-)-Epigallocatechin gallate(EGCG),the most abundant catechin-based flavonoid in green tea leaves,has significant antioxidant,anti-carcinogenic,antimicrobial,and neuroprotective ***,given its low molecular weight and hydrophilic properties,can cross the blood-retinal barrier and is able to reach different ocular tissues such as the lens,cornea,and *** has been shown to provide retinal protection against ischemia;sodium nitroprusside-,N-methyl-D-aspartate-,lipopolysaccharide-,light-,sodium iodate-,or H2 O2-induced damage and diabetic *** suggests that systemic EGCG administration has the potential to protect against retinal degenerative or neurodegenerative diseases such as *** aim of this work was to investigate whether EGCG can protect against RP progression in the animal P23 H line 1,the model of *** P23 H rats were crossed with pigmented Long Evans rats to produce offspring exhibiting the clinical features of *** P23 H rats were treated via intraperitoneal injection with saline or EGCG at a dose of 25 mg/kg every week from P100 to P160 and then compared to wild-type Long Evans *** treated with EGCG showed better visual and retinal electrical function with increased contrast sensitivity and b-wave values compared with those observed in P23 H rats treated with *** reduced lipid peroxidation and increased total antioxidant capacity and catalase and superoxide dismutase *** differences were observed in visual acuity,nitrate levels,nitrite levels or glutathione S-transferase *** conclusion,EGCG not only reduced the loss of visual function in P23 H rats but also improved the levels of antioxidant enzymes and reduced oxidative *** study was approved by the Institutional Animal Care and Use Committee(CE

关键词： antioxidants contrast sensitivity electroretinogram green tea neurodegeneration oxidative stress retinal degeneration retinitis pigmentosa systemic visual acuity

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Combined metabolic activators improve cognitive functions in Alzheimer’s disease patients:a randomised,double-blinded,placebo-controlled phase-II trial

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Translational Neurodegeneration 2023年第1期12卷 849-871页

作者： Burak Yulug Ozlem Altay Xiangyu Li Lutfu Hanoglu Seyda Cankaya Simon Lam Halil Aziz Velioglu Hong Yang Ebru Coskun Ezgi Idil Rahim Nogaylar Ahmet Ozsimsek Cemil Bayram Ismail Bolat Sena Oner Ozlem Ozdemir Tozlu Mehmet Enes Arslan Ahmet Hacimuftuoglu Serkan Yildirim Muhammad Arif Saeed Shoaie Cheng Zhang Jens Nielsen Hasan Turkez Jan Borén Mathias Uhlén Adil Mardinoglu department of Neurology and Neuroscience Faculty of MedicineAlanya Alaaddin Keykubat UniversityAntalyaTurkey Science for Life Laboratory KTH-Royal Institute of TechnologyStockholmSweden department of Neurology Faculty of MedicineIstanbul Medipol UniversityIstanbulTurkey Centre for Host-Microbiome Interaction’s Faculty of DentistryOral and Craniofacial SciencesKing’s College LondonLondonUK department of Women’s and Children’s Health Karolinska InstituteStockholmSweden Functional Imaging and Cognitive-Affective Neuroscience Lab Istanbul Medipol Univer-sityIstanbulTurkey department of Medical pharmacology Faculty of Medi-cineAtatürk UniversityErzurumTurkey department of Pathology Veterinary FacultyAtaturk UniversityErzurumTurkey department of molecular Biology and genetics Faculty of ScienceErzurum Technical UniversityErzurumTur-key School of Pharmaceutical Sciences Zhengzhou UniversityZhengzhouPeople’s Republic of China department of Biology and Biological Engineering Chalmers University of TechnologyGothenburgSweden department of Medical Biology Faculty of MedicineAtatürk UniversityErzurumTurkey department of molecular and Clinical Medicine University of Gothenburg and Sahlgrenska University HospitalGothenburgSweden.

Background Alzheimer’s disease(AD)is associated with metabolic abnormalities linked to critical elements of *** recently administered combined metabolic activators(CMA)to the AD rat model and observed that CMA improves the AD-associated histological parameters in the *** promotes mitochondrial fatty acid uptake from the cytosol,facilitates fatty acid oxidation in the mitochondria,and alleviates oxidative *** Here,we designed a randomised,double-blinded,placebo-controlled phase-II clinical trial and studied the effect of CMA administration on the global metabolism of AD ***-dose CMA included 12.35 g L-serine(61.75%),1 g nicotinamide riboside(5%),2.55 g N-acetyl-L-cysteine(12.75%),and 3.73 g L-carnitine tartrate(18.65%).AD patients received one dose of CMA or placebo daily during the first 28 days and twice daily between day 28 and day *** primary endpoint was the difference in the cognitive function and daily living activity scores between the placebo and the treatment *** secondary aim of this study was to evaluate the safety and tolerability of CMA.A comprehensive plasma metabolome and proteome analysis was also performed to evaluate the efficacy of the CMA in AD *** We showed a significant decrease of AD Assessment Scale-cognitive subscale(ADAS-Cog)score on day 84 vs day 0(P=0.00001,29%improvement)in the CMA ***,there was a significant decline(P=0.0073)in ADAS-Cog scores(improvement of cognitive functions)in the CMA compared to the placebo group in patients with higher ADAS-Cog *** cognitive functions in AD patients were supported by the relevant alterations in the hippocampal volumes and cortical thickness based on imaging ***,the plasma levels of proteins and metabolites associated with NAD+and glutathione metabolism were significantly improved after CMA *** Our results indicate that treatment of AD patients with CMA can lead to enhanced cognit

关键词： Alzheimer’s disease Combined metabolic activators Multi-omics Systems biology Systems medicine

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Fibroblast growth factor-4 and hepatocyte growth factor induce differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocytes

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World Journal of Gastroenterology 2005年第47期11卷 7461-7465页

作者： Xin-Qin Kang Wei-Jin Zang Li-Jun Bao Dong-Ling Li Tu-Sheng Song Xiao-Li Xu Xiao-Jiang Yu department of genetics and molecular Biology Mcclical School of Xi'an Jiaotong UniversityXi'an 710061 Shaanxi Province China department of pharmacology Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education School of Medicine Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China department of Obstetrics and Gynecology First Hospital of Xi'an Jiaotong University Xi'an 710061 Shaanxi Province China

AIM： To investigate the differentiation of human umbilical cord blood （HUCB）-derived mesenchymal stem cells （MSCs） into hepatocytes by induction of fibroblast growth factor-4 （FGF-4） and hepatocyte growth factor （HGF）, and to find a new source of cell types for therapies of hepatic diseases. METHODS： MSCs were isolated by combining gradient density centrifugation with plastic adherence. When HUCB-derived MSCs reached 70% confluence, they were cultured in Iscove modified Dulbecco medium （IMDM） supplemented with 10 mL/L FBS, 20 ng/mL HGF and 10 ng/mL FGF-4. The medium was changed every 4 d and stored for albumin, alpha-fetoprotein （AFP） and urea assay. Expression of CK-18 was detected by immunocytochemistry. Glycogen storage in hepatocytes was determined by PAS staining. RESULTS： By combining gradient density centrifugation with plastic adherence, we could isolate MSCs from 25.6% of human umbilical cord blood. When MSCs were cultured with FGF-4 and HGF, approximately 63.6% of cells became small, round and epithelioid on d 28 by morphology. Compared with the control, the level of AFP increased significantly from d 12 to 18.20：1=1.16 μg/L （t = 2.884, P〈0.05） in MSCs cultured with FGF-4 and HGF, and was higher （54.28±3.11 μg/L） on d 28 （t = 13.493, P〈0.01）. Albumin increased significantly on d 16 （t = 6.68, P〈0.01） to 1.02±0.15 μg/mL, and to 3.63±0.30 μg/mL on d 28 （t = 11.748, P〈0.01）. Urea （4.72±1.03 μmol/L） was detected on d 20 （t = 4.272, P〈0.01）, and continued to increase to 10.28±1.06　μmol/L on d 28 （t = 9.276, P〈0.01）. Cells expressed CK-18 on d 16. Glycogen storage was observed on d 24. CONCLUSION： HUCB-derived MSCs can differentiate into hepatocytes by induction of F-GF-4 and HGF. HUCB derived MSCs are a new source of cell types for cell transplantation therapy of hepatic diseases.

关键词： Mesenchymal stem cell Differentiation Hepatocyte

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Neuroprotective effects of G9a inhibition through modulation of peroxisome-proliferator activator receptor gamma-dependent pathways by miR-128

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Neural Regeneration Research 2024年第11期19卷 2532-2542页

作者： Aina Bellver-Sanchis Pedro AAvila-López Iva Tic David Valle-García Marta Ribalta-Vilella Luis Labrador Deb Ranjan Banerjee Ana Guerrero Gemma Casadesus Coralie Poulard Mercè Pallàs Christian Grinán-Ferré department of pharmacology and Therapeutic Chemistry Institut de Neurociències-Universitat de BarcelonaBarcelonaSpain department of Biochemistry and molecular genetics Feinberg School of MedicineNorthwestern UniversityChicagoILUSA Institute of Biotechnology National Autonomous University of MexicoCuernavacaMexico department of pharmacology and Therapeutics Health Science Center-University of FloridaGainesvilleFLUSA department of Chemistry National Institute of Technology DurgapurM G AvenueDurgapurWest BengalIndia Cancer Research Cancer Lyon Universitéde LyonLyonFrance Inserm U1052 Centre de Recherche en Cancérologie de LyonLyonFrance CNRS UMR5286 Centre de Recherche en Cancérlogie de LyonLyonFrance Centro de Investigación en Red Enfermedades Neurodegenerativas(CIBERNED)Instituto de Salud Carlos IIIMadridSpain Neuroimmunology department National Institute of Neurology and Neurosurgery“Manuel Velasco Suárez”Mexico CityMexico

Dysregulation of G9a,a histone-lysine N-methyltransferase,has been observed in Alzheimer’s disease and has been correlated with increased levels of chronic inflammation and oxidative ***,microRNAs are involved in many biological processes and diseases playing a key role in pathogenesis,especially in multifactorial diseases such as Alzheimer’s ***,our aim has been to provide partial insights into the interconnection between G9a,microRNAs,oxidative stress,and *** better understand the biology of G9a,we compared the global microRNA expression between senescence-accelerated mouse-prone 8(SAMP8)control mice and SAMP8 treated with G9a inhibitor *** found a downregulation of miR-128 after a G9a inhibition treatment,which interestingly binds to the 3′untranslated region(3′-UTR)of peroxisome-proliferator activator receptor γ(PPARG)***,Pparg gene expression levels were higher in the SAMP8 group treated with G9a inhibitor than in the SAMP8 control *** also observed modulation of oxidative stress responses might be mainly driven Pparg after G9a *** confirm these antioxidant effects,we treated primary neuron cell cultures with hydrogen peroxide as an oxidative *** this setting,treatment with G9a inhibitor increases both cell survival and antioxidant ***,up-regulation of PPARγby G9a inhibitor could also increase the expression of genes involved in DNA damage responses and *** addition,we also described that the PPARγ/AMPK axis partially explains the regulation of autophagy markers ***,PPARγ/GADD45αpotentially contributes to enhancing synaptic plasticity and neurogenesis after G9a ***,we propose that pharmacological inhibition of G9a leads to a neuroprotective effect that could be due,at least in part,by the modulation of PPARγ-dependent pathways by miR-128.

关键词： aging cognitive decline epigenetics G9a inhibition microRNAs miR-128 peroxisome-proliferator activator receptorγ(PPARγ) PPARG SAMP8

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Analysis of the combined effect of rs699 and rs5051 on angiotensinogen expression and hypertension

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Chronic Diseases and Translational Medicine 2024年第2期10卷 102-117页

作者： Nicholas R.Powell Tyler Shugg Jacob Leighty Matthew Martin Rolf P.Kreutz Michael T.Eadon Dongbing Lai Tao Lu Todd C.Skaar Division of Clinical pharmacology Department of MedicineSchool of MedicineIndiana UniversityIndianapolisIndianaUSA department of pharmacology and Toxicology School of MedicineIndiana UniversityIndianapolisIndianaUSA department of Cardiology School of MedicineKrannert Institute of CardiologyIndiana UniversityIndianapolisIndianaUSA Division of Nephrology Department of MedicineSchool of MedicineIndiana UniversityIndianapolisIndianaUSA department of Medical and molecular genetics School of MedicineIndiana UniversityIndianapolisIndianaUSA

Background Hypertension(HTN)involves genetic variability in the renin-angiotensin system and influences antihypertensive *** previously reported that angiotensinogen(AGT)messenger RNA(mRNA)is endogenously bound by miR-122-5p and rs699 A>G decreases reporter mRNA in the microRNA functional-assay *** AGT promoter variant rs5051 C>T is in linkage disequilibrium(LD)with rs699 A>G and increases AGT *** independent effect of these variants is understudied due to their LD therefore we aimed to test the hypothesis that increased AGT by rs5051 C>T counterbalances AGT decreased by rs699 A>G,and when these variants occur independently,it translates to HTN-related *** We used in silico,in vitro,in vivo,and retrospective models to test this *** In silico,rs699 A>G is predicted to increase miR-122-5p binding affinity by 3%.Mir-eCLIP results show rs699 is 40–45 nucleotides from the strongest microRNA-binding site in the AGT ***,rs699 A>G increases AGT mRNA in an AGT-plasmid-cDNA HepG2 expression ***-Tissue Expression(GTEx)and UK Biobank analyses demonstrate liver AGT expression and HTN phenotypes are not different when rs699 A>G occurs independently from rs5051 C>***,GTEx and the in vitro experiments suggest rs699 A>G confers cell-type-specific effects on AGT mRNA abundance,and suggest paracrine renal renin-angiotensin-system perturbations could mediate the rs699 A>G associations with *** We found that rs5051 C>T and rs699 A>G significantly associate with systolic blood pressure in Black participants in the UK Biobank,demonstrating a fourfold larger effect than in White *** studies are warranted to determine if altered antihypertensive response in Black individuals might be due to rs5051 C>T or rs699 A>*** like this will help clinicians move beyond the use of race as a surrogate for genotype.

关键词： angiotensinogen genetic and genomic medicine hypertension rs699

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