新型CpGODN对乙型肝炎病毒复制的抑制作用

目的:探讨我室自行设计的CpG ODN(CpG BW001)在体外对乙型肝炎病毒(HBV)复制的抑制作用.方法:CpG BW001刺激人外周血单个核细胞(PBMC)48小时,用VSV病毒保护实验及ELISA法检测培养上清的病毒保护能力及其中的IFN-α含量.将上述CpG BW001刺激人PBMC的上清与HepG2.2.15细胞共孵育12天后收集培养上清,用放射免疫法检测该培养上清液中HBsAg及HBeAg的含量,用点杂交法检测HBV DNA的含量.结果:CpG BW001刺激人PBMC产生以IFN-α为主的抗病毒物质;CpG BW001刺激人PBMC的培养上清作用于HepG2.2.15细胞后,HepG2.2.15细胞培养上清中HBsAg和HBeAg的含量明显减少,且细胞内HBV DNA的含量也显著降低.结论:CpG BW001能通过刺激细胞产生抗病毒物质如IFN-α等,从而在体外抑制HBsAg和HBeAg的表达及HBV的复制.     

CpG-ODN体外抑制乙型肝炎病毒复制的研究

目的:探讨CpG-ODN体外对HBV复制和HBV抗原表达的抑制作用。方法:CpG-ODN体外刺激人外周血单个核细胞(PBMC),用ELISA检测培养液中IFN-α及IFN-γ的水平。将不同比例的PBMC培养上清与HepG2. 2. 15细胞共孵育 2、4、6、8d后,用ELISA和荧光定量PCR法,分别检测培养上清液中HB sAg、HBeAg和HBVDNA的水平。结果:CpG-ODN可诱导PB MC分泌IFN-α和IFN-γ。CpG-ODN本身虽不能直接抑制HBV的复制,但经刺激活的化的PBMC的培养上清,却能显著抑制 2. 2. 15细胞产生HBsAg、HBeAg和HBVDNA,在培养的第 8天,抑制率最高分别达 90. 8%、31. 3%和 32. 2%。结论:CpG-ODN作为一种新型的免疫调节剂,可诱发机体的免疫效应,抑制HBV复制和HBV抗原的表达。     

HMGN2分子体外抗乙型肝炎病毒活性研究

目的： 我们先前的研究证明HMGN2（high mobility group nucleosomal-binding domain 2）是人LAK细胞抗菌的一个新的效应分子,本研究欲检测其体外抗乙型肝炎病毒的活性。方法: 应用反向高效液相色谱技术从人淋巴结组织酸溶性提取物中分离纯化批量HMGN2分子,用质谱精确分子量测定、Western blotting和抗菌试验对分离纯化得到的HMGN2分子进行分子鉴定。采用MTT法检测HMGN2分子对稳定转染复制型HBV重组质粒的肝胚瘤细胞株HepG2.2.15细胞的细胞毒性;用不同浓度HMGN2分子作用于HepG2.2.15细胞,分别在第3 d和第6 d收集细胞培养上清液,ELISA检测上清中HBV表面抗原（HBsAg）及e抗原（HBeAg）,采用实时荧光定量PCR法检测上清液HBV DNA的含量。结果: 从人淋巴结组织中分离纯化获得纯度较高的HMGN2蛋白;在检1-100 mg/L范围内,HMGN2分子对HepG2.2.15细胞无细胞毒性;HMGN2分子在1-5 mg/L水平即可显著抑制HBeAg和HBsAg的表达,可显著降低HBV DNA拷贝数。结论: HMGN2分子在体外具有较强的抗HBV活性。     

注射用免疫核糖核酸II的免疫调节及对顺铂的增效减毒作用

 目的 研究干扰素刺激基因因子3（ISGF3）在干扰素α（IFN-α）抑制乙型肝炎病毒（HBV）复制机制中的作用。 方法 应用定量PCR技术检测IFN-α处理前后人肝癌细胞系HepG2.2.15细胞上清HBV DNA含量，DNA印迹法检测IFN-α处理前后HepG2.2.15细胞内HBV复制中间体的变化，蛋白质印迹法检测IFN-α处理前后HepG2和HepG2.2.15细胞中ISGF3的3个组分即信号转导和转录活化因子（STAT）1、磷酸化STAT2和ISGF3γ以及双链RNA依赖蛋白激酶(PKR)蛋白质表达水平的改变。用酪氨酸酶抑制剂染料木黄酮抑制IFN-α作用后，再次进行上述检测。 结果 IFN-α处理后，HepG2.2.15细胞上清HBV DNA和细胞内HBV复制中间体减少，HepG2和HepG2.2.15细胞中STAT1、磷酸化STAT2、ISGF3γ和PKR蛋白质表达水平均升高。加染料木黄酮抑制IFN-α后，HepG2.2.15细胞上清HBV DNA和复制中间体无减少，而HepG2和HepG2.2.15细胞中STAT1、磷酸化STAT2、ISGF3γ和PKR蛋白质表达均减少。 结论 ISGF3是IFN-α抑制HBV复制的关键因子。     

CpG-ODN活化人NK细胞的初步研究

目的：研究D型CpG-ODN对人免疫细胞的活化效应。方法：CpG-ODN体外刺激人外周血单个核细胞(PBMC)，ELBA检测培养液IFN-α及IFN-γ的含量；RT-PCR检测PBMC中TLR9的表达水平；MTF法观察活化的NK对K562的杀伤作用。结果：CpG-ODN有效诱导PBMC分泌IFN-α和IFN-γ，增强活化的NK细胞对K562细胞的杀伤作用，且能显著上调TLR9 mRNA的表达。结论：在TLR9的介导下，CpG-ODN能有效激活NK细胞，参与机体免疫调节。     

新型融合多肽HTPP-MDC体外抑制HBV复制活性及亚细胞定位

 目的： 研究肝细胞靶向穿膜肽-家蝇天蚕素（HTPP-MDC）融合多肽体外对乙型肝炎病毒（HBV）的抑制作用及其在肝细胞中的定位。方法：不同浓度HTPP-MDC分别与HepG2.2.15细胞和Chang liver细胞共培养,用四甲基偶氮唑盐（MTT）比色法检测药物对细胞的毒性作用,应用ELISA和实时荧光定量 PCR技术定量研究HTPP-MDC对HepG2.2.15细胞系分泌HBsAg、HBeAg及HBV DNA复制的影响。应用激光共聚焦技术研究HTPP-MDC在肝细胞中的定位。结果：HTPP-MDC 在体外能有效抑制HepG2.2.15细胞系分泌HBsAg和HBeAg,对HBV DNA的复制亦有显著抑制作用。激光共聚焦显微镜观察显示HTPP-MDC进入细胞内部。结论：HTPP-MDC在体外对 HBV 复制有较强的抑制作用,并能快速透过细胞膜进入细胞内,有望用于临床治疗慢性乙型肝炎相关疾病。     

霉酚酸对HepG2.2.15细胞中乙型肝炎病毒复制的影响

目的:探讨霉酚酸（MPA）在体外对乙型肝炎病毒(HBV)复制的影响。 方法:将不同浓度的MPA（1-20 mg/L）作用于HepG2.2.15细胞，在外加或不加鸟嘌呤核苷（简称鸟苷）的情况下，分别收集第4 d细胞培养上清，采用酶联免疫吸附试验（ELISA）检测上清中乙型肝炎病毒表面抗原（HBsAg）和e抗原（HBeAg），采用逆转录多聚酶链反应（RT-PCR）检测细胞内乙型肝炎病毒核心蛋白mRNA（HBV core mRNA）,狭缝印迹杂交法定量分析细胞内 HBV DNA。 结果:在不外加鸟苷情况下，MPA对HBV复制具有抑制作用，随着浓度增加，对HBsAg和HBeAg的抑制率逐渐上升，细胞内HBV DNA复制水平下降。外加鸟苷后能逆转MPA对HBV复制的抑制作用。 结论:MPA对HepG2.2.15细胞HBsAg和HBeAg的分泌及HBV DNA复制具有抑制作用。MPA可能通过减少细胞内鸟苷酸的合成来实现对HBV复制的抑制。     

蝉拟青霉多糖体外对乙型肝炎病毒复制的抑制作用

目的: 探讨蝉拟青霉多糖(PcPS)体外对乙型肝炎病毒(HBV)的抑制作用及对HepG2.2.15细胞表达Toll样受体4 (TLR4) mRNA的影响。方法: 以不同浓度PcPS与HepG2.2.15细胞株共培养,以拉米夫定 (LMV)为阳性对照,采用四甲基偶氮唑盐(MTT)法和ELISA法,检测PcPS对HepG2.2.15细胞毒性作用及其分泌HBsAg、HBeAg的情况。实时荧光定量PCR分析PcPS对HepG2.2.15细胞表达与分泌HBV-DNA和TLR4 mRNA表达的影响。结果: 蝉拟青霉多糖在体外能有效抑制HepG2.2.15细胞分泌HBsAg、HBeAg,最大抑制率分别为44.8%、31.0%;对HepG2.2.15细胞内HBV-DNA复制和TLR4 mRNA表达均有一定抑制作用。结论: 一定浓度的蝉拟青霉多糖在体外具有显著抑制HBV的复制作用。     

TWEAK及IFN-γ在诱导表达乙肝病毒的HepG2.2.15细胞凋亡中的协同作用

以转染了HBV全基因组、并稳定表达HBV病毒颗粒的HepG2．2．15细胞为细胞模型，研究新型凋亡分子TWEAK对HBV感染细胞的作用机制。MTT法检测TWEAK和IFN-γ对HepG2．2．15细胞的杀伤效应。流式细胞术(FCM)分析TWEAK和IFN-γ作用后，HepG2．2．15细胞的凋亡率及细胞周期变化。HBsAg和HBeAg ELISA检测试剂盒测定TWEAK与IFN-γ作用24小时后，HBV病毒颗粒的分泌表达状况。TWEAK对于HepG2．2．15细胞有较弱的杀伤效应和诱导凋亡效应，但是与IFN-γ联合作用后，HepG2．2．15细胞的凋亡率显著上调，细胞周期阻滞在G0／G1期，并且TWEAK与IFN-γ能够协同抑制HepG2．2．15细胞病毒颗粒的分泌。TWEAK在IFN-γ的参与下，可以诱导HBV相关的HepG2．2．15细胞发生较强烈的凋亡反应，这提示，在HBV感染者体内的炎症反应环境中，TWEAK极有可能通过与炎性因子的协同效应参与对病毒感染细胞的杀伤效应。     

HBV转染增强细胞因子上调PD-L表达

目的 研究HBV转染的肝细胞系(HepG2.2.15细胞)在细胞因子作用下能否上调PD-L表达.方法 应用肝细胞系(HepG2细胞和HepG2.2.15细胞)为模型,用IL-4、IFN-α、IFN-γ(终质量浓度均为10 ng/ml,刺激时间为12 h)作用于上述细胞系,采用RT-PCR技术检测细胞因子作用前后PD-L表达情况.结果 无论是否转染HBV,IFN-α和IFN-γ均能诱导肝细胞系(HepG2细胞和HepG2.2.15细胞)PD-L1表达;而IL-4不能诱导PD-L1表达,IL-4、IFN-α、IFN-γ均能诱导转染了HBV的HepG2.2.15细胞PD-L2表达,仅IFN-γ能诱导未转染HBV的HepG2细胞PD-L2表达.结论 IFN-α和IFN-γ有较强诱导肝细胞系(HepG2细胞和HepG2.2.15细胞)PD-L1表达上调的作用,HBV在细胞因子诱导HepG2.2.15细胞PD-L2表达中具有促进作用.     

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.     

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.     

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.     

'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.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

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.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.