不同激素剂量和组合对不同品系小鼠超数排卵的影响

背景：超数排卵效果受动物品系、营养水平、年龄、发情周期阶段、光照、超排方法、超排所用激素种类和剂量等诸多因素影响,其中激素剂量和动物品种是关键因素。 目的：探讨不同剂量孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)和人绒毛膜促性腺激素(human chorionic gonadotropin, HCG)组合,对不同品系小鼠超数排卵效果的影响。 方法：分别用不同剂量的PMSG和HCG对ICR鼠、KM鼠和BALB/c鼠进行超数排卵处理,比较激素处理后各品系小鼠超数排卵的胚胎总数、平均胚胎率、正常胚胎及平均可用胚率。 结果与结论：5 IU PMSG+7 IU HCG剂量组合对ICR鼠和KM鼠超排处理效果较好,BALB/c鼠超排的最适激素剂量为3 IU PMSG+5 IU HCG;使用5 IU PMSG+7 IU HCG剂量组合分别处理3种品系小鼠时,ICR鼠和KM鼠平均胚胎率和平均可用胚率显著高于 BALB/c鼠(P < 0.05)。为获得较多的胚胎进行相关实验,应当选择ICR和KM等小鼠进行超排,特别是选择国际通用的ICR鼠。     

早期或晚期补救ICSI对IVF-ET早期胚胎发育和临床妊娠结局的影响

目的研究早期或晚期卵胞浆内单精子注射(ICSI)对体外受精(IVF)后移植前胚胎发育情况和移植后临床妊娠结局的影响,探讨补救ICSI的可行性和优化优化补救ICSI方案,改善体外授精-胚胎移植(IVF-ET)的临床妊娠结局。方法回顾性分析本中心2014年1月1日至2016年1月1日接受IVF-ET的患者的胚胎发育情况和临床妊娠结局,根据授精方式的不同分为ICSI组(A组)、早期补救ICSI组(B组,授精后6-8小时行ICSI)和晚期补救ICSI组(C组,授精18-20小时行ICSI)。对三组患者的基本情况、早期胚胎发育情况和临床妊娠情况分别进行统计分析;根据移植时胚胎发育阶段的不同,将卵裂期胚胎和囊胚分别进行讨论分析。结果 (1)对卵裂期胚胎来说:三组患者的年龄、不孕年限、Gn用量、基础FSH、移植日内膜厚度、获卵数、移植胚胎数、卵裂率和冷冻胚胎率均无统计学差异(P0.05),但三组的受精率、D3优质胚胎率、D3可用胚胎率有非常显著的统计学差异(P≤0.001),而且三组的临床妊娠率有显著统计学差异(P=0.003);(2)对囊胚移植来说:三组组患者的年龄、不孕年限、Gn用量、基础FSH、移植日内膜厚度、获卵数、移植胚胎数和卵裂率没有统计学差异(P0.05),但三组的受精率、D3优质胚胎率、D3可用胚胎率和可用囊胚率存在非常显著的统计学差异(P≤0.001),且三组的临床妊娠率存在统计学差异(P=0.046)。结论早期补救ICSI相比较于晚期补救ICSI,可以获得较好的胚胎发育和临床妊娠结局,但都逊色于普通ICSI。补救ICSI能获得相当的可用胚胎和临床妊娠,可以作为常规IVF受精失败或受精较差的补充,而且早期补救ICSI要优于晚期补救ICSI。     

内毒素对去卵透明带2-细胞小鼠胚胎体外发育的影响

目的 观察不同剂量浓度内毒素对去卵透明带2—细胞小鼠胚胎体外发育的影响。方法 获取小鼠2—细胞胚胎，用胰蛋白酶法去除卵透明带后，分别置于含有0，1pg/ml，5pg/ml和10pg/ml内毒素的CZB液滴中培养，此外，用蛋白酶法去除卵透明带，在不含内毒素的CZB液滴中培养，观察胚胎体外发育情况。结果 用胰蛋白酶法去卵透明带的2—细胞小鼠胚胎，其囊胚率随着培养液中内毒素剂量的增加而降低，且5pg/ml和10pg/ml时与对照组(不含内毒素)相比差异显著(P＜0．001)。在有内毒素存在时，停滞在2—细胞或4细胞期的胚胎彼此分离排成列，不再呈球形。部分卵裂球出现空泡或碎裂。另外，用蛋白酶法去除透明带的2—细胞鼠胚的囊胚率为61．7％，显著低于胰蛋白酶法的73．8％(P＜0．001)。结论 微量内毒素即明显抑制去卵透明带2—细胞小鼠胚胎的体外发育，并表现出剂量效应；去卵透明带2—细胞小鼠胚胎试验可应用于培养液的微量内毒素检测；胰酶法去除卵透明带后胚胎的囊胚率优于蛋白酶法。     

环孢霉素A对小鼠妊娠失败模型妊娠预后的影响

目的 :探讨环孢霉素A(CyclosporinA ,CsA)对小鼠妊娠失败模型及正常妊娠模型妊娠预后的影响。方法 :于孕 4d分别给小鼠妊娠失败模型及正常妊娠模型腹腔注射不同剂量 (0 ,5 ,10 ,15mg kg)CsA ,于孕 14d处死各组小鼠 ,观察各实验组胚胎吸收率、胎鼠及胎盘重量变化。结果 :①CsA可使小鼠妊娠失败模型胚胎吸收率显著降低 ,接近于小鼠正常模型胚胎吸收率 ;同时胎鼠及胎盘重量无显著性差异 ,但有增加趋势。②CsA对小鼠正常妊娠模型胚胎吸收率无明显影响 ;胎鼠及胎盘重量亦无显著性差异。结论 :于孕早期 (胚胎着床期 )腹腔注射CsA可使小鼠妊娠失败模型胚胎吸收率恢复至正常水平 ,从而可能成为保胎的潜在药物。     

输卵管积水对小鼠胚胎体外发育能力的影响

目的探讨输卵管积水对小鼠胚胎体外发育能力的影响。方法收集人输卵管积水，小鼠促超排卵，收集2细胞胚胎和囊胚，随机分配到含有不同浓度输卵管积水的培养液中，观察胚胎的发育，计算成囊率，囊胚孵出率。结果输卵管积水组的胚胎成囊率和囊胚孵出率低于无积水组，并呈剂量依赖性。结论输卵管积水能显著影响胚胎的早期发育潜能。     

ATF4基因在小鼠胚胎围着床期子宫内膜的表达

目的 探讨转录激活因子4(ATF4)在小鼠胚胎围着床期子宫内膜中的表达规律及在胚胎植入过程中的作用.方法 妊娠0～7d小鼠子宫内膜,用RT-PCR、免疫组织化学法和Western blot分别检测小鼠内膜ATF4 mRNA及蛋白的表达;用子宫角内注射ATF4抗体进行干预.结果 1)妊娠各组小鼠子宫内膜ATF4 mRNA的表达量均显著高于未孕组d0(P ＜0.0S),且随妊娠天数的增加其表达量逐渐增高,到d4达到峰值,之后逐渐下降;2)ATF4蛋白主要在基质细胞胞质表达,其表达规律与mRNA表达规律基本一致.3)子宫角ATF4抗体注射后与对照组相比着床数明显减少.结论 ATF4在小鼠妊娠早期可能参与子宫内膜蜕膜化过程,有利于着床.     

小鼠胚胎与子宫内膜中整合素α_5的表达

目的:观察整合素α5在小鼠早期胚胎和子宫内膜中的表达分布状况,探讨整合素在胚胎着床中的可能作用。方法:用免疫组织化学ABC法染色,对发育不同时期的小鼠胚卵和妊娠2 d～6 d、8 d、13 d以及非妊娠期的子宫内膜中整合素α5的表达分布进行形态学观察与半定量分析。结果:整合素α5的阳性反应物质在小鼠早期胚卵胞浆内;非妊娠期子宫内膜和妊娠期的脱膜上皮与基质细胞内均有整合素α5表达,植入前表达相对较弱,植入后随妊娠的进展逐渐增强。结论:提示整合素α5在小鼠胚卵着床中起重要作用:推测它不参与胚卵对母体子宫内膜的识别和粘附,而是参与了粘附后的侵入及胚胎发育过程。     

小鼠胚胎与子宫内膜中整合素α5的表达

目的：观察整合素α5在小鼠早期胚胎和子宫内膜中的表达分布状况，探讨整合素在胚胎着床中的可能作用。方法：用免疫组织化学ABC法染色，对发育不同时期的小鼠胚卵和妊娠2d-6d、8d、13d以及非妊娠期的子宫内膜中整合素内的表达分布进行形态学观察与半定量分析。结果：整合素α5的阳性反应物质在小鼠早期胚卵胞浆内；非妊娠期子宫内膜和妊娠期的脱膜上皮与基质细胞内均有整合素α5表达，植入前表达相对较弱，植入后随妊娠的进展逐渐增强。结论：提示整合素α5在小鼠胚卵着床中起重要作用：推测它不参与胚卵对母体子宫内膜的识别和粘附，而是参与了粘附后的侵入及胚胎发育过程。     

小鼠非同步胚胎移植技术

目的:比较小鼠非同步胚胎移植技术。方法:将小鼠不同发育阶段的着床前胚胎非同步移植到不同假孕受体小鼠的输卵管或子宫内,观察非同步胚胎移植对小鼠胚胎移植成功率和后代出生重的影响。结果:CD-1小鼠受精卵、2-细胞胚胎、桑椹胚和囊胚都可移植到假孕0.5d受体小鼠输卵管继续发育,妊娠受体在移植后19d出生小鼠,胚胎移植出生率分别为56.3%、62.5%、59.4%和58.3%。桑椹胚和囊胚也可移植到假孕2.5 d受体子宫继续发育,妊娠受体在移植后17 d出生小鼠胚胎移植出生率分别为57.5%和62.5%。囊胚移植假孕0.5 d受体输卵管的出生小鼠的出生重显著大于移植假孕2.5 d受体子宫的出生小鼠。结论:不同发育阶段的胚胎若移植相同的假孕受体,则妊娠受体有相同的妊娠期;相同发育阶段的胚胎若移植不同的假孕受体,则妊娠受体有不同的妊娠期,假孕时间短的受体有较长的妊娠期和显著增加的小鼠出生重。     

小鼠胚泡植入早期FucT-Ⅶ和sLe(X)寡糖抗原表达

目的探讨妊娠第1～5天(d1～5)小鼠子宫内膜唾液酸化的路易斯寡糖[sLe(X)]合成关键酶α-1,3岩藻糖基转移酶基因(FucT-Ⅶ)的表达和其表面sLe(X)寡糖抗原表达对胚胎植入的影响。方法应用RT-PCR和免疫组化方法检测妊娠第1～5天小鼠子宫内膜FucT-Ⅶ及寡糖抗原的表达规律。用子宫角内注射sLe(X)单克隆抗体的方法检测小鼠胚胎着床数。结果妊娠第1～5天小鼠子宫组织均有FucT-Ⅶ的表达,且在妊娠d4表达更强(P<0.05)。妊娠第1～5天sLe(X)寡糖抗原表达在子宫内膜的腔上皮和腺上皮。单侧子宫角sLe(X)抗体注射后胚胎的着床数量明显较对侧(注射等体积生理盐水)减少。结论在小鼠胚胎着床过程中,FucT-Ⅶ及sLe(X)寡糖抗原可能参与了胚泡的定位、黏附及侵袭过程,在胚泡植入的早期发挥作用。     

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.     

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.     

'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.     

Hypo- und Hypermagnesämie aus kardiologischer Sicht

Zusammenfassung Eine Reihe pathologischer Zustände bedingen Magnesiummangel. Zustände mit Hypermagnesämie sind ebenfalls bekannt, doch wesentlich seltener. Für den Kardiologen beachtenswert ist, daß unter Therapie mit bestimmten Diuretica bei Herzinsuffizienz, bei Herzinfarkt, Kardiomyopathie, Digitalisintoxikation und bestimmten Herzrhythmusstörungen Hypomagnesämie beobachtet wurde. Leider kann in der klinischen Routine nur ein extracelluläres Magnesiumdefizit durch Serumbestimmungen gemessen werden; über Magnesiummangel einzelner Organe kann nichts ausgesagt werden. Hinweise für Magnesiummangel geben aber neben der Messung des Serumspiegels Anamnese, klinischer Befund, bestimmte EKG-Veränderungen wie auch evtl. Hypokalämie, ein Zustand, bei dem sich oft — besonders bei Aldosteronismus — parallele Veränderungen zeigten.Tierexperimente deuten darauf hin, daß infarktähnliche Läsionen unter Magnesiummangel entstehen, doch ob Herzinfarkt beim Menschen durch Magnesiummangel ausgelöst werden kann, ist noch ungeklärt. In Leichenherzen zeigte sich im Infarktgebiet neben Calciumakkumulation signifikanter Magnesiumverlust, wobei unklar blieb, ob sich Ursache oder Folge des Infarktes widerspiegelten. Falls ein ursächlicher Zusammenhang besteht, ist er im Myokardstoffwechsel selbst zu suchen, wie bei der Alkoholkardiomyopathie, wo myokardialer Magnesiummangel zumindest als pathogenetischer Teilfaktor anerkannt wird. Andererseits versucht man aber auch Beziehungen zwischen Atherosklerose, Blutgerinnung und Hypomagnesämie herzustellen, in der Meinung, daß Magnesiummangel auch über den coronaren Pathomechanismus des Herzinfarktes wirken könnte. Sicher scheint, daß gewisse EKG-Veränderungen und Herzrhythmusstörungen durch einen irritierten Magnesiumhaushalt bedingt sein können, da sie bei Gabe bzw. Entzug von Magnesium verschwinden. Daß Magnesiummangel die Glykosidtoleranz verringert, wird tierexperimentell bestätigt. Unter Hypomagnesämie bewirkt Acetylstrophanthidin eher und länger Rhythmusstörungen als ohne, außerdem lassen diese sich durch Magnesiumgaben eliminieren. Da in gewissen Fällen spontane und digitalisinduzierte Herzrythmusstörungen durch Magnesiuminjektionen beseitigt wurden, scheint Magnesium als Therapeuticum angebracht. Einsatz verschiedener Magnesiumsalze bei Angina pectoris, degenerativen Herzerkrankungen und Herzinsuffizienz ohne geprüften und offensichtlich gestörten Magnesiumhaushalt ist fragwürdig, weil keine eindeutigen klinischen Erfolgsbeweise vorliegen. Immerhin mag es aber larvierte, durch Serumbestimmungen nicht erfaßbare Mangelzustände geben. Allgemein erscheint es aus kardiologischer Sicht ratsam, den Magnesiumhaushalt zu überwachen und in entsprechenden Fällen auszugleichen, um möglichen Myokardläsionen oder fatalen Herzrhythmusstörungen entgegenzuwirken.     

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.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.