组织特异性启动子在溶瘤腺病毒中的应用

溶瘤腺病毒（Oncolytic Adenovirus,Ad）是肿瘤基因治疗的研究热点。利用肿瘤或组织特异性启动子（Tissue—specific promoter,TSP）控制腺病毒关键基因的表达、实现腺病毒在肿瘤细胞特异性复制是目前溶瘤腺病毒构建的主要策略。另外,负载治疗基因可以增强杀灭肿瘤细胞的效果,     

溶瘤病毒与肿瘤

溶瘤病毒治疗肿瘤是一种全新的肿瘤基因治疗方法,利用病毒自身特性发现并杀伤肿瘤细胞,近年来受到人们广泛关注。溶瘤病毒包括细小病毒、呼肠孤病毒、新城疫病毒、水疱性口炎病毒、麻疹病毒、腺病毒、疱疹病毒和甲型流感病毒等。目前已经有多个溶瘤病毒产品进入抗肿瘤临床试验阶段,且疗效较为显著。本文对该领域最新研究进展作一概述。     

溶瘤病毒与肿瘤

溶瘤病毒治疗肿瘤是一种全新的肿瘤基因治疗方法,利用病毒自身特性发现并杀伤肿瘤细胞,近年来受到人们广泛关注.溶瘤病毒包括细小病毒、呼肠孤病毒、新城疫病毒、水疱性口炎病毒、麻疹病毒、腺病毒、疱疹病毒和甲型流感病毒等.目前已经有多个溶瘤病毒产品进入抗肿瘤临床试验阶段,且疗效较为显著.本文对该领域最新研究进展作一概述.     

溶瘤病毒的临床应用

新城疫病毒（newcastle disease virus，NDV）、单纯疱疹病毒-1（herpes simplex virus-1，HSV-1）、呼肠孤病毒（reovirus）、溶瘤腺病毒（oncolytic adenovirus）等是由嗜肿瘤特性而被用来改造成溶瘤病毒，它特异性识别并感染肿瘤细胞，最终导致细胞溶胀而摧毁肿瘤细胞，但无法在正常机体细胞内复制而不具有杀伤作用，理论上具有更高的抗肿瘤效应和更低的副作用。文章就以上溶瘤病毒的国内外临床应用现状及目前仍面临的主要问题进行综述。     

溶瘤病毒应用于肿瘤治疗的研究进展

溶瘤病毒应用于肿瘤治疗是近 10年来兴起的一种肿瘤治疗的新方案 ,现已进行了临床试验研究 ,其结果令人鼓舞。本文详细介绍几种溶瘤病毒包括溶瘤腺病毒、溶瘤疱疹病毒、呼 (吸道 )肠 (道 )病毒、新城疫病毒等治疗肿瘤的最新进展 ,为溶瘤病毒应用于肿瘤治疗过程提供新的启示。     

放射性碘标记特异性溶瘤重组腺病毒KH901在荷人肝癌裸鼠体内的抑瘤作用

研究131I标记的特异性溶瘤重组腺病毒KH901的生物活性及131I-KH901对荷人肝细胞肝癌裸鼠肿瘤的抑制作用。采用N-溴代琥珀酰亚胺(NBS)法用131I标记KH901,并采用ELISA法测定粒细胞-巨噬细胞集落刺激因子(GM-CSF)的生物活性;通过给药后荷人肝细胞肝癌裸鼠肿瘤的生长实验观察131I-KH901对肿瘤生长的影响,组织切片观察肿瘤组织形态学变化。结果显示:131I-KH901在肿瘤细胞中表达大量的GM-CSF因子,24 h后,在肿瘤细胞和正常细胞中产生的GM-CSF因子分别为183.27±6.90 pg/ml和20.44±0.77 pg/ml;肿瘤生长观察结果显示,131I-KH901瘤内给药能明显抑制肿瘤生长,抑瘤率为71.3%,显著大于131I组(22.7%)及KH901瘤内给药组(52.7%)(P<0.05),组织形态学观察示,注射131I-KH901后,肿瘤出现大片坏死区。因此,131I-KH901能明显抑制裸鼠体内人肝癌细胞的生长,可望成为腺病毒溶瘤和放射性核素联合治疗肿瘤的新型药物。     

特异性溶瘤腺病毒调控肺癌细胞表达PD-L1及诱导凋亡的研究

目的:探究特异性溶瘤腺病毒(Ad-Mock、Ad-T)对肺癌细胞上PD-L1表达的调控作用及对细胞凋亡的影响.方法:采用荧光定量PCR法及Western blot检测溶瘤腺病毒在24 h和48 h对NCI-H446、NCI-H226、A549细胞表达PD-L1的影响;通过结晶紫染色、WST-1、Hoechst与JC-1...     

溶瘤腺病毒靶向治疗脑胶质瘤的研究现状及进展

脑胶质瘤是常见的颅内恶性肿瘤之一,手术治疗及术后放、化疗仍是其主要的治疗手段,因其术后复发率高、放化疗副作用及耐药性等问题,无法达到令人满意的治疗效果。基因治疗作为目前发展迅速的精准治疗措施之一,递送载体是其发挥治疗作用的关键因素,溶瘤腺病毒因其特异性、高效性及安全性等优势,使其在多种肿瘤的基础研究及临床应用中被作为基因递送的载体。该文主要介绍了溶瘤腺病毒在脑胶质瘤中的研究现状,联合治疗的策略及临床应用最新进展,以期为溶瘤腺病毒在脑胶质瘤的靶向治疗中提供一定的理论依据。     

荷载溶瘤病毒细胞载体的研究进展

溶瘤病毒的给药方式分瘤内注射和静脉注射。静脉注射溶瘤病毒可被免疫系统完全排除而不能发挥其抗肿瘤作用。应用具有淋巴细胞特性、干细胞特性、肿瘤细胞特性的细胞载体荷载并运输溶瘤病毒到达肿瘤部位发挥作用,可更好地发挥抗免疫/肿瘤效应。     

表达GFP基因的溶瘤腺病毒对大肠癌细胞的体外杀伤作用

目的： 研究在端粒酶启动子驱动下表达绿色荧光蛋白（GFP）及 5型腺病毒早期基因（E1A）基因的腺病毒Ad/hTERT-GFP-E1对大肠癌细胞的杀伤作用及其可能的机制。方法:将不同滴度Ad/hTERT-GFP-E1感染大肠癌及正常细胞,以巨细胞病毒（CMV）启动子驱动下表达GFP基因的腺病毒Ad/CMV-GFP作为载体对照,通过半数组织培养感染量（TCID50）法测定病毒滴度及体外复制能力,然后用MTT法及细胞克隆形成试验评价该病毒在体外对肿瘤细胞及正常细胞的杀伤作用,并对病毒感染后的细胞进行原位凋亡检测。结果:Ad/hTERT-GFP-E1能够在DLD1细胞内持续复制,GFP的表达能够对病毒的感染和复制起到监测作用;MTT结果显示该病毒对于结直肠肿瘤细胞有显著杀伤作用,而对于正常细胞没有明显的杀伤作用;对病毒感染后的DLD1进行细胞凋亡检测发现Ad/hTERT-GFP-E1所引起的凋亡率显著高于对照组(P＜0.01)。结论:溶瘤腺病毒Ad/hTERT-GFP-E1能够选择性地在肿瘤细胞内复制并杀伤肿瘤细胞,其机制与细胞凋亡的途径有关。     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

Class II HLA in Georgia Caucasus Tbilisi Georgians and their Mediterranean ancestry: The Usko Mediterranean languages

Georgia (or Sakartvelo in its own language) is a South Caucasus Mts. country with its easternmost part is enigmatically named Iberia, like the Iberian Peninsula, which may refer to rivers “Kura” and “Ebro” or their valleys respectively. Most of their inhabitants speak Georgian which is included within Dene-Caucasian group and Usko-Mediterranean subgroup of languages. The latter includes Basque, Berber, ancient Iberian-Tartessian, Etruscan, Hittite, Minoan Lineal A and others. In the present paper, HLA class II -DRB1 and -DQB1 alleles has been studied and extended haplotypes calculated. Most frequent haplotypes are also of Mediterranean origin (i. e.: (A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*51)-DRB1*13:01-DQB1*06:03, or (A*24-B*35)-DRB1*01:01-DQB1*05:01) and DA genetic distances show that closest world populations to Georgians are Mediterraneans. Georgians also show common extended haplotypes ((A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*13)-DRB1*07:01-DQB1*02:01 and (A*03-B*35)-DRB1*11:01-DQB1*03:01) with Svan people, a secluded population in North Georgia mountains. We can conclude that Georgians belong to a very old Mediterranean substratum according to both linguistics (Usko Mediterranean languages) and HLA genetics.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.