小鼠精子3种活体染色法的比较分析

目的 比较小鼠精子伊红、锥兰和苯胺黑-伊红3种活体染色法的存活率,结合活动率,低渗肿胀-伊红结合(HOS-EY)试验评价小鼠精子存活率的方法.方法 取昆明系小鼠(n=15)附睾尾精子制成精子悬液,分别应用0.15%伊红、0.25%锥兰和10%苯胺黑-1.0%伊红3种活体染色法检测精子存活率,同时采用精子活动率和HOS-EY试验作对照测试.结果 小鼠精子用3种活体染色法测得的存活率没有显著差异(P＞0.05),存活率与活动率有直线回归关系(P＜0.05),HOS-EY试验中头-尾膜完整的精子率低子活动率和存活率(P＜0.05).结论 伊红、锥兰和苯胺黑-伊红3种活体染色法均可应用于小鼠精子存活率检测,结合检测精子活动率和HOS-EY试验可以进一步反映小鼠精子的存活状态.     

应用人精子头-尾膜完整性试验对生育力组及不育组精液的评价

目的应用人精子头-尾膜完整性试验,比较生育力组与不育组男性精液4种精子膜完整性类型的差异.方法精液标本分为生育力组(n=32)和不育组(n=50),采用"低渗肿胀-伊红拒染"结合试验,检测人精子头-尾膜完整性.结果头膜尾膜均损伤的Ⅰ型精子和头膜损伤-尾膜完整的Ⅲ型精子,生育组和不育组存在明显差异(P<0.01);头膜完整-尾膜损伤的Ⅱ型精子,生育组和不育组无明显差异(P>0.05);头膜-尾膜均完整的Ⅳ型精子,生育组明显高于不育组(P<0.01);Ⅳ型精子率与生育组精子活动率(n=32, r=0.82,P<0.01)和不育组精子活动率(n=50,r=0.80,P<0.01)均呈显著相关性.结论生育组具有较多头膜-尾膜均完整的精子."低渗肿胀-伊红拒染"结合试验能够清晰显示包括过渡型膜损伤的4种膜完整性类型的精子,精子头-尾膜完整性可以作为评价精子生理学的一项检测指标.     

厨师职业与男性不育

目的探讨厨师职业对男性不育症患者精液质量的影响。方法对67例厨师不育者和364例不育对照组,采用计算机辅助分析系统对男性精液进行精子密度,精子活动率、精子活力等方面进行全面分析,同时采用伊红染色法检测精子存活率。结果厨师职业组与对照组比较,精液量、p H值差异无统计学意义(P0.05),而精子密度、存活率、精子活率差异具有统计学意义(P0.05),两组精子活力之间差异具有统计学意义(P0.01)。结论厨师可影响精液质量,主要表现在精子密度、活动率和活力降低,其原因可能与厨师所处的职业环境有关。     

衰老对小鼠精子成熟和受精能力的影响

目的：探讨衰老对附睾精子成熟、体外受精和胚胎发育的影响。方法： 取老龄小鼠（18月龄，n=15）和青年小鼠（6月龄，n=15）附睾头精子和附睾尾精子，分别检测精子密度、存活率、活动率、正常形态率和胞浆小滴率，并通过体外受精比较附睾尾精子受精率和各阶段胚胎发育率。结果：老龄小鼠附睾头精子和附睾尾精子活动率、精子密度显著低于青年组（P<0.01），胞浆小滴率和畸形精子率显著高于青年组（P<0.05），附睾尾精子的受精率下降（P<0.01），胚胎各阶段发育率降低（P<0.01）。结论：衰老影响小鼠精子功能及附睾精子成熟过程。小鼠可作为雄性生殖衰老研究的动物模型。     

精液中生精细胞5种染色方法的探讨与评价

对脱落于精液中的生精细胞，用瑞－吉染色法可使细胞核着色清晰度好；用伊红－瑞－吉染色法能区别死亡和活的精子，并使精子顶体着色清晰；用苯胺黑－伊红－吉染色法精子核染成红色，精子鞭毛不着色；用煌绿－瑞－吉染色法，成熟精子着色浅，不成熟精子头部着紫红色；过氧化物酶－瑞－吉混合染色，中性白细胞核呈紫红色，胞浆呈浅绿色。５种染色方法中瑞－吉法对生精细胞染色效果最好，可视精液检查的不同要求选用不同方法。     

精子膜的完整性与男性不育的相关性研究

目的探讨精子膜完整性与精子功能的相关性。方法收集140例精液标本，其中生育组50例，不育组90例，采用伊红-Y水试验，低渗肿胀试验，检测精子形态和精子膜低渗肿胀率。结果生育组和不育组比较，精子头部未着色率、低渗肿胀率有显著性差异（P〈0．01）；同时，精子膜低渗肿胀率与精子活率密切相关，精子低渗肿胀率越高，其活率也越高（P〈0．01）。结论精子膜功能可能与精液参数密切相关，精子膜功能是评价男性生育力的重要指标之一。     

精子密度、存活率、活动力、形态与宫腔内人工授精妊娠结局

目的探讨精子密度、存活率、活动力（a＋b），形态与宫腔内人工授精妊娠结局的相关性。方法选自以男性不育为因素289对夫妇进行IUI周期治疗：在授精日，取男方精液进行计算机综合分析，常规方法授精后，将其精液检测结果与妊娠结局相比较。结果精子密度、存活率、活力（a＋b）％、形态与IUI妊娠结局概率分别是：15．2％，16．3％，16．9％，18．7％；精子密度组：（30～60）×10^6／ml与（60～90）X10^6／ml、≥100×10^6／ml、≤20X10^6／ml比较有差异性P〈0．01；存活率组：30％-60％与60％～80％、≥80％比较有差异性P〈0．01；活力组：30％～45％与10％～20％、20％～30％、〉50％组间比较有差异性P〈0．Ol；形态组：〈15％纽与a〉30％与30％≥A≥15％比较有差异性P〈0.01。结论精子密度、存活率、活力（a＋b）％、形态与IUI妊娠结局密切相关，可以作为评价宫腔内人工授精的一项实验室指标；精子形态与IUI妊娠结局呈正相关；精子较低{密度、活率、活力（a＋b）％}与IUI妊娠结局呈正相关；单纯性男性少、弱精子症在IUI周期中能获得较高的妊娠率。     

一氧化氮与白细胞精子症不育的关系探讨

目的　探讨一氧化氮 (NO)与白细胞精子症不育的关系。方法　参照WHO标准方法 ,进行精液常规分析。采用过氧化物酶染色法检测精液中白细胞密度。用改良的低渗肿胀法检测精子细胞膜的完整性。采用镀铜镉还原荧光法 ,检测精液中NO代谢产物硝酸盐 (NO 3 )。结果　白细胞精子症不育组白细胞的密度为 (1.4 8± 0 .90 )× 10 9/L ,NO水平为 (10 3.5± 2 .0 ) μmol/L ,显著高于正常生育组的(0 .73± 0 .2 8)× 10 9/L和 (41.6± 1.8) μmol/L (P分别为<0 .0 5和 <0 .0 0 1)。精子的活动率、精子速度试验 (SVT)及精子头、尾部膜完整率 ,均明显低于生育组 (P <0 .0 1) ,而精子的死亡率则显著高于生育组 (P <0 .0 1)。结论　NO水平与白细胞密度呈正相关 ;与精子的活动率、SVT及精子头、尾膜的完整率呈负相关。表明当精液中的白细胞增高时 ,可产生过量的NO导致精子中毒受损使精子的受精能力下降     

miR-338-3p通过负性调节cPKCγ表达减轻小鼠神经元缺血损伤

目的评估miR-338-3p对氧糖剥夺(OGD)/复糖复氧(R)神经元损伤的影响及其机制。方法建立小鼠脑中动脉阻塞(MCAO)和神经元氧糖剥夺(OGD)模型;Western blot检测1 h MCAO/R 24、48和72 h小鼠脑梗死区周围皮质(n=5)和1 h OGD/R 0、6、12和24 h神经元(n=5)cPKCγ蛋白表达量;实时定量PCR检测cPKCγ及miR-338-3p mRNA水平;生物信息学分析cPKCγ3′UTR与miR-338-3p结合位点;噻唑兰(MTT)比色法和乳酸脱氢酶(LDH)法测定神经元(n=6)的存活率和凋亡率。采用免疫荧光与高内涵细胞成像分析技术(HCA)检测caspase-3蛋白表达情况。结果 1 h MCAO/R 24、48和72 h小鼠脑梗死区周围皮层和1 h OGD/R 0、6、12和24 h神经元中cPKCγ蛋白及mRNA水平均明显上升(P0.001);cPKCγ3′UTR存在1个miR-338-3p结合位点,miR-338-3p与cPKCγmRNA水平呈相反趋势;过表达miR-338-3p后1 h OGD/R 24 h神经元存活率下降(P0.001),调亡率上升(P0.001),caspase-3蛋白平均荧光强度值增高(P0.001),抑制miR-338-3p后1 h OGD/R 24 h神经元存活率上升(P0.001),调亡率下降(P0.001),caspase-3蛋白平均荧光强度值降低(P0.001)。结论 miR-338-3p通过负性调节cPKCγ表达而使神经元存活率上升,凋亡率下降,减轻缺血神经元损伤。     

圆头精子的冷冻保存

目的:探讨圆头精子的冷冻保存效果及其精子功能改变。方法:6例圆头精子症患者各提供1-3份精液标本,应用不含卵黄冷冻保护剂和二步降温方法冷冻保存圆头精子症标本,检测冷冻精子活动率、头-尾膜完整率、存活时间和顶体蛋白酶活性。结果:圆头精子标本冷冻保存后,精子活动率显著减少,膜完整率显著下降,头膜损伤-尾膜完整的精子率显著升高(n=13,P＜0.01)。体外存活时间缩短。精子冷冻前后无顶体蛋白酶活性。不含卵黄冷冻保护剂与含卵黄冷冻保护剂,以及两步冷冻与多步冷冻的效果比较未见显著差异(n=7,P＞0.05)。结论:圆头精子可经历冷冻保存后存活,但冷冻复苏率低。冷冻-解冻过程显著降低精子功能。不含卵黄冷冻保护剂和两步降温可应用于冷冻保存圆头精子症标本。     

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.