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Gene Targeting by Homology-Directed Repair in Rice Using a Geminivirus-Based CRISPR/Cas9 System

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Molecular Plant 2017年第7期10卷 1007-1010页

作者： Mugui Wang Yuming Lu jose ramon Botella Yanfei Mao Kai Hua Jian-kang Zhu Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai 201602 China Plant Genetic Engineering Laboratory School of Agriculture and Food Sciences University of Queensland Brisbane QLD 4072 Austraria Department of Horticulture and Landscape Architecture Purdue University West Lafayette IN 47907 USA

Dear Editor, Rice （Oryza sativa） is the staple food for more than half of the world＇s population. Technologies enabling precise and efficient DNA knock-in or replacement, hereinafter referred to as KI, have the pot... 详细信息

关键词： DNA REPLICONS GENOME

TML:a language to specify aerial robotic missions for the framework Aerostack

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International Journal of Intelligent Computing and Cybernetics 2017年第4期10卷 491-512页

作者： Martin Molina ramon A.Suarez-Fernandez Carlos Sampedro jose Luis Sanchez-Lopez Pascual Campoy Department of Artificial Intelligence Technical University of Madrid(UPM)MadridSpain Centre for Automation and Robotics Technical University of MadridMadridSpain

Purpose–The purpose of this paper is to describe the specification language TML for adaptive mission plans that the authors designed and implemented for the open-source framework Aerostack for aerial robotics.Design/methodology/approach–The TML language combines a task-based hierarchical approach together with a more flexible representation,rule-based reactive planning,to facilitate adaptability.This approach includes additional notions that abstract programming details.The authors built an interpreter integrated in the software framework Aerostack.The interpreter was validated with flight experiments for multi-robot missions in dynamic environments.Findings–The experiments proved that the TML language is easy to use and expressive enough to formulate adaptive missions in dynamic environments.The experiments also showed that the TML interpreter is efficient to execute multi-robot aerial missions and reusable for different platforms.The TML interpreter is able to verify the mission plan before its execution,which increases robustness and safety,avoiding the execution of certain plans that are not feasible.Originality/value–One of the main contributions of this work is the availability of a reliable solution to specify aerial mission plans,integrated in an active open-source project with periodic releases.To the best knowledge of the authors,there are not solutions similar to this in other active open-source projects.As additional contributions,TML uses an original combination of representations for adaptive mission plans(i.***.task trees with original abstract notions and rule-based reactive planning)together with the demonstration of its adequacy for aerial robotics.

关键词： Adaptive mission plans Aerial robotic systems Autonomous aerial systems Mission plan specification

Cancer and diet: significance of diet in a patient whose life expectancy was 4 to 5 months

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Cancer Advances 2023年第2期6卷 1-8页

作者： jose ramon Toro Lopez Independent Researcher Sevilla 41500Spain

Clinical summary:The patient,62 years old,a professional laboratory technician specialist,married with two children who in the month of January 2017,came to the emergency department for presenting problems of metrorrhagia and severe pain in left hip that prevented her from ambulation.Once the appropriate tests were performed,the clinical judgment was moderately differentiated endometroid adenocarcinoma with squamous metaplasia infiltrating the cervix,M1(bone and lung metastases);image compatible with left femoral DVT and PET in the right lower lobar branch,and her life expectancy at that time was four to five months.Objective:Given the situation in which the patient was,the few posibilties of a restorative treatment,and the short life expectancy expected for the patient,the only objective was to seek help through a special diet focusing the interest mainly on the nutritional aspect so that it could help to control especially the metastases,the tumor cell and the activation and reinforcement of the immune system.Method:In 2017,when the study was started,a systematic literature review was conducted in search of information related to diets,cancer and metastasis.In this study,the references used for the establishment of the diet have been taken into account and updated to the last five years,where we have been able to confront and confirm the same results obtained in 2017.For the reviews,databases related to health sciences on published articles that related alkaline diets with cancer and metastasis have been used.Results:The patient was recommended a special alkaline dissociated diet based mainly on a nutritional point of view that she started from the first day of treatment with the intention that she could be controlled and favor her recovery;after six months,both tumors and metastases and the problems of DVT and PTE had practically disappeared.Conclusions:The results(although it is only one case)make us think about nutrition's importance,especally in this type of disease.We know this disease is not specific to a

关键词： diet cancer,alkaline dissociated diet alkaline diet dissociated diet minerals metastasis collagen

Endoscopic ultrasound guided vascular access and therapy:A promising indication

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World Journal of Gastrointestinal Endoscopy 2010年第6期2卷 198-202页

作者： Enrique Vazquez-Sequeiros jose ramon Foruny Olcina University Hospital ramon y Cajal

Endoscopic ultrasound(EUS)is an imaging technique that has consolidated its role as an important tool for diagnosis and therapeutics.In recent years we have seen a dramatic increase in the number of EUS-guided therapeutic indications(celiac plexus neurolysis/block, pseudocyst drainage,etc).Preliminary reports have suggested EUS may also be used to guide vascular access for both imaging and treating different vascular diseases.This review aims to objectively describe the existing evidence in the field.

关键词： Endosonography Fine needle aspiration Endoscopic ultrasound-FNA Bleeding Vascular therapy

A detailed procedure for CRISPR/Cas9-mediated gene editing in Arabidopsis thaliana

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Science Bulletin 2015年第15期60卷 1332-1347页

作者： Wenshan Liu Xiaohong Zhu Mingguang Lei Qingyou Xia jose ramon Botella Jian-Kang Zhu Yanfei Mao School of Life Sciences Chongqing University Shanghai Center for Plant Stress Biology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences State Key Laboratory of Silkworm Genome Biology Southwest University Department of Horticulture and Landscape Architecture Purdue University School of Agriculture and Food Sciences University of Queensland

The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesis and transgenic methods to study gene function and improve crop traits. CRISPR/Cas facilitates targeted gene editing through RNA-guided DNA cleavage followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. Here we describe a detailed procedure for our previously developed and highly efficient CRISPR/Cas9 method that allows the generation of heritable-targeted gene mutations andcorrections in Arabidopsis. This protocol describes the strategies and steps for the selection of targets, design of single-guide RNA(sg RNA), vector construction and analysis of transgenic lines. We also offer a method to target two loci simultaneously using vectors containing two different sg RNAs. The principles described in this protocol can be applied to other plant species to generate stably inherited DNA modifications.

关键词： RNA编辑基因组拟南芥转基因载体 DNA裂解介导程序协议描述

Social media in cardiology:Reasons to learn how to use it

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World Journal of Cardiology 2019年第10期11卷 217-220页

作者： Rafael Vidal-Perez jose Juan Gómez de Diego Julia Grapsa Ricardo Fontes-Carvalho jose ramon Gonzalez-Juanatey Heart Failure Unit Cardiology DepartmentHospital Clinico Universitario de SantiagoSantiago de Compostela 15706A CorunaSpain Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares SpainSantiago de Compostela 15706A CorunaSpain Cardiovascular Institute Hospital Universitario San CarlosMadrid 28040Spain Cardiology Department St Bartholomew HospitalBarts Health TrustLondon EC1A 7BEUnited Kingdom Cardiology Department Centro Hospitalar GaiaUniversity of PortoPorto 4434-502Portugal

Social media has changed the way we learn,educate,and interact with our peers.The dynamic nature of social media and their immediate availability through our portable devices(smartphones,tablets,smartwatches,etc.)is quickly transforming the way we participate in society.The scope of these digital tools is broad as they deal with many different aspects:Teaching and learning,case discussion,congresses coverage,peer to peer interaction,research are examples worth mentioning.The scientific societies considered more innovative,are promoting these tools between their members.These new concepts need to be known by the cardiologists to stay updated,as countless information is moving rapidly through these channels.We summarize the main reasons why learning how to use these tools to be part of the conversation is essential for the cardiologist in training or fully stablished.

关键词： Social media Cardiology Congress Learning Teaching Interaction Cardiovascular diseases Impact Factor Portable devices Smartphone Tablet

New therapeutic strategy with extracorporeal membrane oxygenation for refractory hepatopulmonary syndrome after liver transplant: A case report

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World Journal of Transplantation 2024年第1期14卷 210-214页

作者： Belinda Sánchez Pérez María Pérez Reyes jose aranda Narvaez Julio Santoyo Villalba jose Antonio Perez Daga Claudia Sanchez-Gonzalez Julio Santoyo-Santoyo Hepatobiliary and Trasplantation Unit General and Digestive Surgery DepartmentUniversity Regional HospitalMalaga 29010Spain Department of General and Digestive Surgery Regional University Hospital of MalagaMalaga 29010Spain

BACKGROUND Due to the lack of published literature about treatment of refractory hepatopulmonary syndrome(HPS)after liver transplant(LT),this case adds information and experience on this issue along with a treatment with positive outcomes.HPS is a complication of end-stage liver disease,with a 10%-30%incidence in cirrhotic patients.LT can reverse the physiopathology of this process and restore normal oxygenation.However,in some cases,refractory hypoxemia persists,and extracorporeal membrane oxygenation(ECMO)can be used as a rescue therapy with good results.CASE SUMMARY A 59-year-old patient with alcohol-related liver cirrhosis and portal hypertension was included in the LT waiting list for HPS.He had good liver function(Model for End-Stage Liver Disease score 12,Child-Pugh class B7).He had pulmonary fibrosis and a mild restrictive respiratory pattern with a basal oxygen saturation of 82%.The macroaggregated albumin test result was>30.Spirometry demonstrated a forced expiratory volume in one second(FEV1)of 78%,forced vital capacity(FVC)of 74%,FEV1/FVC ratio of 81%,diffusion capacity for carbon monoxide of 42%,and carbon monoxide transfer coefficient of 57%.He required domiciliary oxygen at 2 L/min(16 h/d).The patient was admitted to the intensive care unit(ICU)and extubated in the first 24 h,needing high-flow therapy and non-invasive ventilation and inhaled nitric oxide afterwards.Reintubation was needed after 72 h.Due to the non-response to supportive therapies,installation of ECMO was decided with progressive recovery after 9 d.Extubation was possible on the tenth day,maintaining a high-flow nasal cannula and de-escalating to conventional oxygen therapy after 48 h.He was discharged from ICU on postoperative day(POD)20 with a 90%-92%oxygen saturation.Steroid recycling was needed twice for acute rejection.The patient was discharged from hospital on POD 27 with no symptoms,with an 89%-90%oxygen saturation.CONCLUSION Due to the favorable results observed,ECMO could become the central axis of treatment of HPS and ref

关键词： Liver transplantation Hepatopulmonary syndrome Refractory hypoxemia Treatment Extracorporeal membrane oxygenation Case report

Current status and potential of morphometric sperm analysis

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Asian Journal of Andrology 2016年第6期18卷 863-870页

作者： Alejandro Maroto-Morales Olga Garcia-Alvarez Manuel ramon Felipe Martinez-Pastor M Rocio Fernandez-Santos A josefa Soler jose Julian Garde SaBio IREC (CSIC - UCLM -JCCM) Albacete Spain Biomedical Center Medical Faculty in Pilsen Charles University in Prague Pilsen Czech Republic Regional Center of Animal Selection and Reproduction (CERSYRA) JCCM Valdepenas Spain 4Institute for Animal Health and Cattle Development University of Leon LeOn Spain Department of Molecular Biology University of Leon Leon Spain Faculty of Pharmacy (UCLM) Albacete Spain

The spermatozoon is the most diverse cell type known and this diversity is considered to reflect differences in sperm function. How the diversity in sperm morphology arose during speciation and what role the different specializations play in sperm function, however, remain incompletely characterized. This work reviews the hypotheses proposed to explain sperm morphological evolution, with a focus on some aspects of sperm morphometric evaluation; the ability of morphometrics to predict sperm cryoresistance and male fertility is also discussed. For this, the evaluation of patterns of change of sperm head morphometry throughout a process, instead of the study of the morphometric characteristics of the sperm head at different stages, allows a better identification of the males with different sperm cryoconservation ability. These new approaches, together with more studies employing a greater number of individuals, are needed to obtain novel results concerning the role of sperm morphometry on sperm function. Future studies should aim at understanding the causes of sperm design diversity and the mechanisms that generate them, giving increased attention to other sperm structures besides the sperm head. The implementation of scientific and technological advances could benefit the simultaneous examination of sperm phenotype and sperm function, demonstrating that sperm morphometry could be a useful tool for sperm assessment.

关键词： computer-assisted sperm morphometric analysis mammals sperm function sperm morphometry

Topical Applications of Thiosulfinate-Enriched Allium sativum Extract Accelerates Acute Cutaneous Wound Healing in Murine Model

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Chinese Journal of Integrative Medicine 2020年第11期26卷 812-818页

作者： Juan Luis Santiago Eva Maria Galan-Moya jose ramon Munoz-Rodriguez Miguel Angel de la Cruz-Morcillo Francisco Javier Redondo-Calvo Ignacio Gracia-Fernandez Peter M.Elias jose Manuel Perez-Ortiz Mao-Qiang Man Dermatology Service Hospital General Universitario de Ciudad RealCiudad Real(13004)Spain Translational Research Unit Hospital General Universitario de Ciudad RealCiudad Real(13004)Spain Translational Oncology Laboratory Centro Regional de Investigaciones BiomedicasUniversidad de Castilla-La ManchaAlbacete(02008)Spain Instituto de Tecnologia Quimica y Medioambiental Universidad de Castilla-La ManchaCiudad Real(13004)Spain Dermatology Service Vetera ns Affairs Medical Centerand Department of DermatologyUniversity of CaliforniaSan FranciscoCA(94121)USA Dermatology Hospital Southern Medical UniversityGuangzhou(510091)China

Objective To determine whether topical applications of thiosulfinate-enriched Allium sativum extract(TASE)can accelerate acute cutaneous wound healing(WH)in a murine model.Methods Keratinocyte viability and in vitro wound closure were assessed in keratinocyte cultures.Effects of topical TASE(0.5μg/mL of allicin in 97%ethanol)on acute cutaneous WH were determined in a murine model of acute cutaneous wound.Twelve mice were alternately assigned to the vehicle-and TASE-treated groups(n=6 per group).Expression levels of mRNA for keratinocyte differentiation marker-related proteins(filaggrin,loricrin and involucrin)and lipid synthetic enzymes(elongation of very long chain fatty acids protein 4(ELOVL4),fatty acid synthase(FA2H),3-hydroxy-3-methyl-glutaryl-coenzyme A reductase(HMGCoA),and serine palmitoyltransferase(SPT))were assessed using real-time quantitative polymerase chain reaction on day 3 and 8 after wounding,while transepidermal water loss(TEWL)rates were measured in wounded areas.Results TASE accelerated WH both in vivo(40%vs.22%reduction in wound area,P<0.01)and in vitro(90%vs.65%reduction in wound area,P<0.01).Moreover,topical applications of TASE upregulated the expression levels of epidermal mRNA for ELOVL4,HMGCoA,SPT,filaggrin,loricrin and involucrin(P<0.05 vs.vehicle-treated controls)on day 3 after wounding.Likewise,TASE significantly lowered TEWL rates in comparison with vehicle alone on day 8(33.06±2.09 g/(m^2·h)vs.24.60±2.04 g/(m^2·h),P<0.01).Conclusions Topical applications of TASE stimulated keratinocyte proliferation and formation of epidermal permeability barrier function,leading to acceleration of acute cutaneous WH.Topical products containing TASE could be used to manage acute cutaneous WH.

关键词： Allium sativum thiosulfinate wound healing permeability barrier keratinocytes proliferation