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Developmental staging of male murine embryonic gonad by SAGE analysis

Journal of Genetics and Genomics 2009年第4期36卷 215-227页

作者： Tin-Lap Lee Yunmin Li Diana Alba Queenie r Vong Shao-Ming Wu Vanessa Baxendale Owen M. rennert Yun-Fai Chris Lau Wai-Yee Chan Section on Developmental Genomics Laboratory of Clinical Genomics Eunice Kennedy Shriver National Institute of Child Health and Human Development National Institutes of Health Bethesda MD 20892 USA depar.ment of Medicine VA Medical Center University of California San Francisco CA 94121 USA depar.ment of Pediatrics Biochemistry Molecular & Cellular Biology Georgetown University Washington DC 20007 USA

Despite the identification of key genes such as Sry integral to embryonic gonadal development, the genomic classification and identification of chromosomal activation of this process is still poorly understood. To better understand the genetic regulation of gonadal development, we performed Serial Analysis of Gene Expression （SAGE） to profile the genes and novel transcripts, and an average of 152,000 tags from male embryonic gonads at E10.5 （embryonic day 10.5）, E11.5, E12.5, E13.5, E15.5 and E17.5 were analyzed. A total of 275,583 non-singleton tags that do not map to any annotated sequence were identified in the six gonad libraries, and 47,255 tags were mapped to 24,975 annotated sequences, among which 987 sequences were uncharacterized. Utilizing an unsupervised pattern identification technique, we established molecular staging of male gonadal development. rather than providing a static descriptive analysis, we developed algorithms to cluster the SAGE data and assign SAGE tags to a corresponding chromosomal position; these data are displayed in chromosome graphic format. A prominent increase in global genomic activity from E10.5 to E17.5 was observed. Important chromosomal regions related to the developmental processes were identified and validated based on established mouse models with developmental disorders. These regions may represent markers for early diagnosis for disorders of male gonad development as well as potential treatment targets.

关键词： SAGE transcriptome male gonads gene tag novel transcripts cluster analysis chromosome transcription hotspot

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The use of high resolution anterior segment optical coherence tomography for the characterization of conjunctival lymphoma, conjunctival amyloidosis and benign reactive lymphoid hyperplasia

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Eye and Vision 2019年第1期6卷 123-130页

作者： Nandini Venkateswaran Carolina Mercado Ann Q.Tran Armando Garcia Pedro Francisco Monsalve Diaz Sander r.Dubovy Anat Galor Carol L.Karp Bascom Palmer Eye Institute University of Miami Miller School of Medicine900 NW 17th StreetMiamiFL 33136USA depar.ment of Ophthalmology Miami Veterans Administration Medical CenterMiamiFLUSA

Background:Conjunctival lymphoma,conjunctival amyloidosis and benign reactive lymphoid hyperplasia(BrLH)are conditions that often have a similar appearance on the ocular *** use of high resolution anterior segment optical coherence tomography(Hr-OCT)enables clinicians to evaluate distinctive differences in tissue morphology and cellular patterns in various ocular surface *** this study,we characterize the morphological differences seen in conjunctival lymphoma,conjunctival amyloidosis and BrLH on Hr-OCT ***:A retrospective chart review was performed of patients with biopsy proven conjunctival lymphoma,conjunctival amyloidosis and BrLH between 2012 and 2019 at the Bascom Palmer Eye *** were excluded if Hr-OCT imaging was not performed on initial ***:Thirty-four total eyes of 27 patients were *** eyes had conjunctival lymphoma(16 patients),8 eyes had conjunctival amyloidosis(6 patients)and 6 eyes had BrLH(5 patients).All conditions appeared clinically as pink,red or yellow subepithelial lesions but had different features on *** lymphoma,Hr-OCT images typically showed homogenous,dark subepithelial lesions with smooth borders,containing monomorphic dot-like ***-OCT images of amyloidosis typically showed heterogeneous,dark lesions with irregular borders,often containing hyperreflective linear ***-OCT images of BrLH showed variable infiltration of the subepithelial tissue,at times with homogenous lesions containing dot-like infiltrates like lymphoma and other times with more hyperreflective,subepithelial *** cytometry and gene rearrangement was needed for final differentiation between BrLH and lymphoma ***:Distinctive features on Hr-OCT of conjunctival lymphoma,conjunctival amyloidosis and BrLH can help characterize these lesions beyond what is apparent with the clinical *** studies can further validate this technology’s use with m

关键词： High resolution anterior segment optical coherence tomography Ocular surface imaging Ocular surface lesions Conjunctival lymphoma Conjunctival amyloidosis Benign reactive lymphoid hyperplasia

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Extra-skeletal manifestations in mice affected by Clcn7-dependent autosomal dominant osteopetrosis type 2 clinical and therapeutic implications

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Bone research 2019年第2期7卷 195-209页

作者： Antonio Maurizi Mattia Capulli Annabel Curle rajvi Patel Argia Ucci Juliana Alves Cotes Harriet Oxford Shireen r. Lamandé John F. Bateman Nadia rucci Anna Teti depar.ment of Biotechnological and Applied clinical Sciences University of L’AquilaL’AquilaItaly Murdoch Children’s research Institute and univer.ity of Melbourne MelbourneAustralia

Autosomal dominant osteopetrosis type 2 (ADO2) is a high-density brittle bone disease characterized by bone pain,multiple fractures and skeletal-related events,including nerve compression syndrome and hematological *** demonstrated that in mice carrying the heterozygous Clcn7^G213r mutation,whose human mutant homolog CLCN7^G215r affects patients,the clinical impacts of ADO2 extend beyond the skeleton,affecting several other *** hallmark of the extra-skeletal alterations is a consistent perivascular fibrosis,associated with high numbers of macrophages and lymphoid *** clinical information in a small cohort of patients confirms extra-skeletal alterations consistent with a systemic disease,in line with the observation that the CLCN7 gene is expressed in many ***2 mice also show anxiety and depression and their brains exhibit not only perivascular fibrosis but also β-amyloid accumulation and astrogliosis,suggesting the involvement of the nervous system in the pathogenesis of the ADO2 extra-skeletal ***-skeletal organs share a similar cellular pathology,confirmed also in vitro in bone marrow mononuclear cells and osteoclasts,characterized by an impairment of the exit pathway of the Clcn7 protein product,ClC7,through the Golgi,with consequent reduced ClC7 expression in late endosomes and lysosomes,associated with high vesicular pH and accumulation of autophagosome ***,an experimental sirNA therapy,previously proven to counteract the bone phenotype,also improves the extra-skeletal *** results could have important clinical implications,supporting the notion that a systematic evaluation of ADO2 patients for extra-skeletal symptoms could help improve their diagnosis,clinical management,and therapeutic options.

关键词： Extra-skeletal manifestations dominant osteopetrosis implications

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Isolation and identification of adipose-derived stromal/stem cells from breast cancer patients after exposure neoadjuvant chemotherapy

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World Journal of Experimental Medicine 2020年第3期10卷 26-40页

作者： Ashleigh rapp Hagaman Ping Zhang Kiavash r Koko ryan S Nolan Marc W Fromer John Gaughan Martha Matthews depar.ment of Surgery Cooper University HospitalCamdenNJ 08103United States Cooper Medical School of rowan univer.ity CamdenNJ 08103United States

BACKGrOUND With recent research advances,adipose-derived stromal/stem cells(ASCs)have been demonstrated to facilitate the survival of fat grafts and thus are increasingly used for reconstructive procedures following surgery for breast ***,in patients,following radiation and chemotherapy for breast cancer suggest that these cancer treatment therapies may limit stem cell cellular functions important for soft tissue wound *** clinical translation to patients that have undergone cancer treatment,it is necessary to understand the effects of these therapies on the ASC's ability to improve fat graft survival in clinical *** To investigate whether the impact on ASCs function capacity and recovery in cancer patients may be due to the *** ASCs were isolated from the cancerous side and noncancerous side of the breast from the same patients with receiving neoadjuvant chemotherapy(NAC)or notreceiving *** were in vitro treated with 5-fluorouracil(5-FU),doxorubicin(DXr),and cyclophosphamide(Cytoxan)at various *** stem cells yield,cell viability,and proliferation rates were measured by growth curves and MTT *** capacity for adipogenesis was determined by qPCr analysis of the specific gene markers and histological *** No significant differences were observed between the yield of ASCs in patients receiving NAC treatment and not-receiving *** yield from the cancerous side of the breast showed lower than the noncancerous side of the breast in both patients receiving NAC and not-receiving *** proliferation rates of ASCs from patients didn’t differ much before and after NAC upon in vitro culture,and these cells appeared to retain the capacity to acquire adipocyte traits simile to the ASCs from patients not-receiving *** cessation and washout of the drugs for another a week of culturing,ASCs showed a slow recovery of cell growth capacity in 5-FU-treated groups but was not

关键词： Human adipose-derived stromal/stem cells Breast cancer Neoadjuvant chemotherapy Chemotherapeutic agents Adipogenic differentiation Proliferation

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Synergistic inhibition of SCr1- and ErBB2-driven brain metastatic breast cancer cells

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Journal of Cancer Metastasis and Treatment 2019年第3期4卷 59-72页

作者： Shahnaz r.Ansari Zain Jandial Xiwei Wu Xueli Liu Mike Y.Chen Khairul I.Ansari Division of Neurosurgery Beckman Research InstituteCity of HopeDuarteCA 91010USA depar.ment of Molecular and cellular Biology Beckman Research InstituteCity of HopeDuarteCA 91010USA Division of Biostatistics Beckman Research InstituteCity of HopeDuarteCA 91010USA

Aim: Metastasis to the brain has become a major limitation to the life expectancy and quality of life for many patients with breast cancer. Unfortunately, other than radiation and palliative treatments with trastuzumab, and pertuzumab, no effective therapy for brain metastases is currently available. This study seeks to identify novel gene targets and pharmaceutical Intervention against breast cancer brain metastasis. Methods: The detailed methods applied to this study, including comparative rNA sequencing and bioinformatics analysis of sequence data, ingenuity pathway analysis, protein-protein interaction analysis, high throughput screening of clinical and pre-clinical drugs, cell viability and proliferation assay, toxicity and apoptosis assay using fluorescence-activated cell sorting, real-time PCr, western blotting, statistical analysis of data. results: The study reveals critical roles for SrC, ErBB2, PIK3CA, and GABA in the proliferation and survival of breast cancer brain metastatic (BBM) cells and showed that SrC- and ErBB2-mediated activation of PIK3-AKT/mTOr signaling regulates BBM cell survival. Selective inhibition of these candidate genes alone or in combination induces robust apoptosis in BBM cells Conclusion: The findings of this study provide a rationale for further preclinical evaluation of SrC-targeting regimens in combination with ErBB2 inhibitors and/or GABA agonists to target breast cancer brain metastasis.

关键词： Breast cancer brain metastasis human epidermal growth factor receptor 2 ErBB2 SrC astrocytes PI3KCA

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实体肿瘤外周血细胞免疫功能实验室检测专家共识

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中华检验医学杂志 2023年第12期46卷 1235-1248页

作者：中国医师协会检验医师分会北京医师协会医学检验专科医师(技师)分会国家癌症中心国家肿瘤区域医疗中心国家医学检验临床医学研究中心(中国医科大学附属第一医院) 尚红崔巍中国医师协会检验医师分会北京医师协会医学检验专科医师(技师)分会国家癌症中心国家肿瘤区域医疗中心中国医科大学附属第一医院国家医学检验临床医学研究中心

外周血细胞免疫功能检测在实体肿瘤免疫治疗受益者筛选、免疫相关不良反应监测、肿瘤疗效及预后评估等方面受到临床关注。为促进临床有效选择和解读实体肿瘤诊疗相关的外周血细胞免疫功能指标,中国医师协会检验医师分会、北京医师协会... 详细信息

外周血细胞免疫功能检测在实体肿瘤免疫治疗受益者筛选、免疫相关不良反应监测、肿瘤疗效及预后评估等方面受到临床关注。为促进临床有效选择和解读实体肿瘤诊疗相关的外周血细胞免疫功能指标,中国医师协会检验医师分会、北京医师协会医学检验专科医师(技师)分会、国家癌症中心、国家肿瘤区域医疗中心、国家医学检验临床医学研究中心(中国医科大学附属第一医院)组织相关领域的学术专家,对外周血细胞免疫功能指标包括T淋巴细胞亚群精细分型、髓源性抑制细胞和细胞因子的临床应用价值进行了充分总结,并撰写了专家共识,同时对外周细胞免疫功能实验室检测指标和方案或方法给出了推荐和参考方向,以期为实体肿瘤患者的精准诊疗提供帮助。

关键词：肿瘤外周血细胞免疫功能流式细胞术免疫治疗共识

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The Effect of Antineoplastons A10 and AS2-1 and Metabolites of Sodium Phenylbutyrate on Gene Expression in Glioblastoma Multiforme

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Journal of Cancer Therapy 2014年第10期5卷 929-945页

作者： Stanislaw r. Burzynski Sonali S. Patil depar.ment of Cell and Molecular Biology Burzynski Research Institute Inc. Stafford TX USA

Antineoplastons are peptide and amino acid derivatives that occur naturally in the human body. They inhibit the growth of neoplastic cells without growth inhibition of normal cells. Phenylacetylglutaminate (PG) is an active ingredient of antineoplastons A10 and AS2-1 (ANP) and is also a metabolic by-product of phenylbutyrate (PB). The formulation of antineoplaston AS2-1 is a 4:1 mixture of phenylacetate (PN) and PG. Antineoplaston A10 is a 4:1 mixture of PG and isoPG. This study investigates the molecular mechanism of action of PG and PN. The Human U87 glioblastoma (GBM) cell line was used as the model system in this study. A total human gene array screen using the Affymetrix Human Genome plus 2.0 oligonucleotide arrays was performed using mrNA derived from U87 cells exposed to PG and PN. Pathway analysis was performed to allow the visualization of effect on metabolic pathways and gene interaction networks. Our preliminary results indicate that PG and PN interrupt signal transduction in rAS/MAPK/ErK and PI3K/AKT/PTEN pathways, interfere with cell cycle, decrease metabolism and promote apoptosis in human U87 GBM cells. The effect on multiple cellular pathways and targets, suggests that ANP and PB are promising candidates for clinical studies in GBM.

关键词： Glioblastoma Antineoplastons Phenylbutyrate Gene Expression Pathways Cell Cycle

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residual Cardiovascular risk—Is Inflammation the Primary Cause?

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World Journal of Cardiovascular Diseases 2018年第1期8卷 59-69页

作者： David S. Schade r. Philip Eaton Division of endocrinology and Metabolism Department of Internal Medicine School of Medicine University of New Mexico Al-buquerque NM USA

Every cardiovascular clinical trial that has examined the beneficial effects of lowering LDL cholesterol to prevent cardiovascular events has demonstrated residual cardiovascular risk in the interventional treatment group. residual risk is the term applied to the cardiovascular events (e.g., myocardial infarction, stroke, and cardiovascular death) that occur in spite of being on “optimal” medical therapy. This term is usually applied to secondary intervention studies,?i.e., lipid lowering treatments in subjects who have already had at least one cardiovascular event. Studies that described residual risk have attributed it, at least in part, to the fact that the LDLc has not been lowered sufficiently to stop atherosclerotic plaque formation and rupture into the arterial lumen. However, a recent cardiovascular intervention clinical trial which achieved a very low group median LDLc of 30 mg/dl still demonstrated significant residual risk. Of more importance to reducing residual risk may be addressing the ongoing inflammation in the coronary arteries that results in cellular liberation of cytokines and proteases that attack the atherosclerotic plaque’s fibrous cap. recent studies have shown that inflammation may act independently of LDL to cause cardiovascular events. This article provides evidence that inflammation is the primary cause of residual risk and will need to be treated as aggressively as LDL lowering if CVD events in the post treatment period are to be significantly reduced. Addressing major risk factors including obesity, diabetes, smoking, hypertension and hyperlipidemia are critical to reducing inflammation. Statins and aspirin are the mainstay medications to reduce ongoing inflammation. However, newer pharmaceuticals may also be required to reduce inflammation to undetectable levels. Targeting inflammation to eradicate residual cardiovascular risk will be the next therapeutic challenge facing primary care physicians.

关键词： residual risk Inflammation Cholesterol C-reactive Protein Cardiovascular Events

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CTLA4 protects against maladaptive cytotoxicity during the differentiation of effector and follicular CD4^(+)T cells

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cellular & Molecular Immunology 2023年第7期20卷 777-793页

作者： Yuwei Hao Bahar Miraghazadeh rochna Chand Ainsley r.Davies Chelisa Cardinez Kristy Kwong Morgan B.Downes rebecca A.Sweet Pablo F.Cañete Lloyd J.D’Orsogna David A.Fulcher Sharon Choo Desmond Yip Geoffrey Peters Sonia Yip Matthew J.Witney Maxim Nekrasov Zhi-Ping Feng David C.Tscharke Carola G.Vinuesa Matthew C.Cook Centre for Personalised Immunology John Curtin School of Medical ResearchThe Australian National UniversityCanberraACTAustralia Translational research unit The Canberra HospitalCanberraACTAustralia Division of Immunology and Infectious Diseases John Curtin School of Medical ResearchThe Australian National UniversityCanberraACTAustralia depar.ment of Immunology Fiona Stanley HospitalPerthWAAustralia depar.ment of Immunology The Royal Children’s HospitalMelbourneVICAustralia depar.ment of Medical Oncology The Canberra HospitalCanberraACTAustralia ANU Medical School The Australian National UniversityCanberraACTAustralia NHMrC clinical Trials unit The University of SydneySydneyNSWAustralia The ACrF Biomolecular resource Facility John Curtin School of Medical ResearchThe Australian National UniversityCanberraACTAustralia ANU Bioinformatics Consultancy John Curtin School of Medical ResearchThe Australian National UniversityCanberraACTAustralia Francis Crick Institute 1 Midland RdLondon NW11ATUK Cambridge Institute of Therapeutic Immunology and Infectious Disease University of CambridgeCambridgeUnited Kingdom.

As chronic antigenic stimulation from infection and autoimmunity is a feature of primary antibody deficiency(PAD),analysis of affected patients could yield insights into T-cell differentiation and explain how environmental exposures modify clinical phenotypes conferred by single-gene ***57 marks dysfunctional T cells that have differentiated after antigenic ***,while circulating CD57^(+)CD4^(+)T cells are normally rare,we found that they are increased in patients with PAD and markedly increased with CTLA4 haploinsufficiency or *** performed single-cell rNA-seq analysis of matched CD57^(+)CD4^(+)T cells from blood and tonsil *** CD57^(+)CD4^(+)T cells(CD4cyt)exhibited a cytotoxic transcriptome similar to that of CD8^(+)effector cells,could kill B cells,and inhibited B-cell ***4 restrained the formation of *** CD57 also marked an abundant subset of follicular helper T cells,which is consistent with their antigen-driven differentiation,this subset had a preexhaustion transcriptomic signature marked by TCF7,TOX,and ID3 expression and constitutive expression of CTLA4 and did not become cytotoxic even after CTLA4 ***,CD57^(+)CD4^(+)T-cell cytotoxicity and exhaustion phenotypes are compartmentalised between blood and germinal ***4 is a key modifier of CD4^(+)T-cell cytotoxicity,and the pathological CD4cyt phenotype is accentuated by infection.

关键词： CTLA4 Immunodeficiency Cell exhaustion Terminal differentiation Cytotoxic CD4^(+)T cells CD57

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Safety,immunogenicity,and protection provided by unadjuvanted and adjuvanted formulations of a recombinant plant-derived virus-like particle vaccine candidate for COVID-19 in nonhuman primates

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cellular & Molecular Immunology 2022年第2期19卷 222-233页

作者： Stéphane Pillet Prabhu SArunachalam Guadalupe Andreani Nadia Golden Jane Fontenot Pyone Pyone Aye Katharina röltgen Gabrielle Lehmicke Philipe Gobeil Charlotte Dubé Sonia Trépanier Nathalie Charland Marc-AndréD’Aoust Kasi russell-Lodrigue Christopher Monjure robert V.Blair Scott D.Boyd rudolf P.Bohm Jay rappaport François Villinger Nathalie Landry Bali Pulendran Brian J.Ward Medicago Inc. QuébecQCCanada. Institute for Immunity Transplantation and InfectionStanford University School of MedicineStanford UniversityStanfordCAUSA. Tulane National Primate research Center CovingtonLAUSA. New Iberia research Center University of Louisiana at LafayetteNew IberiaLAUSA. depar.ment of Pathology Stanford University School of MedicineStanford UniversityStanfordCAUSA. depar.ment of Microbiology and Immunology Tulane University School of MedicineNew OrleansLAUSA. depar.ment of Microbiology and Immunology Stanford University School of MedicineStanford UniversityStanfordCAUSA. Institute for Immunity Transplantation&InfectionStanford University School of MedicineStanford UniversityStanfordCAUSA research Institute of the McGill univer.ity Health Centre MontrealQCCanada.

Although antivirals are important tools to control severe acute respiratory syndrome coronavirus 2(SArS-CoV-2)infection,effective vaccines are essential to control the current coronavirus disease 2019(COVID-19)***-derived virus-like particle(VLP)vaccine candidates have previously demonstrated immunogenicity and efficacy against ***,we report the immunogenicity and protection induced in rhesus macaques by intramuscular injections of a VLP bearing a SArS-CoV-2 spike protein(CoVLP)vaccine candidate formulated with or without Adjuvant System 03(AS03)or cytidine-phospho-guanosine(CpG)*** a single dose of the unadjuvanted CoVLP vaccine candidate stimulated humoral and cell-mediated immune responses,booster immunization(at 28 days after priming)and adjuvant administration significantly improved both responses,with higher immunogenicity and protection provided by the AS03-adjuvanted *** micrograms of CoVLP adjuvanted with AS03 induced a polyfunctional interleukin-2(IL-2)-driven response and IL-4 expression in CD4 T *** were challenged by multiple routes(i.e.,intratracheal,intranasal,and ocular)with a total viral dose of 106 plaque-forming units of *** viral replication in nasal swabs and bronchoalveolar lavage fluid(BALF)as well as fewer SArS-CoV-2-infected cells and immune cell infiltrates in the lungs concomitant with reduced levels of proinflammatory cytokines and chemotactic factors in the BALF were observed in animals immunized with the CoVLP adjuvanted with *** clinical,pathologic,or virologic evidence of vaccineassociated enhanced disease was observed in vaccinated *** CoVLP adjuvanted with AS03 was therefore selected for vaccine development and clinical trials.

关键词： SArS-CoV-2 Virus-like particles Non-humane primates AS03 CpG1018

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