黄嘌呤抗视神经切断后视网膜细胞凋亡的作用

目的　探讨眼球玻璃体注射 3 异丁基 1 甲基 (IBMX)对切断视神经后视网膜细胞凋亡的作用。方法　用荧光核染料 332 5 8和琼脂糖凝胶电脉技术观察切断视神经 5、7和 1 4d后成年金黄地鼠视网膜细胞凋亡的变化。结果　 ( 1 )视神经切断 5、7、1 4d后 ,节细胞层 (GCL)细胞凋亡密度 (cell/mm2 )分别为 94 60± 2 9 40、81 2 0± 1 5 1 9和 31 2 0± 1 1 43,给予IBMX组在上述时间点凋亡密度为 67 80± 1 3 83、60 2 0± 9 68和 2 7 0 0± 9 5 7。IBMX 7d组与对照组比较有显著差异 (P <0 0 5 )。 ( 2 )琼脂糖凝胶电脉显示切断 7d的视网膜细胞DNA抽提物显示典型的DNA梯形条带。其它各间间点 ( 5d和 1 4d)以及玻璃体内注射IBMX均未见典型DNA梯形条带。结论　IBMX有抗视神经切断后视网膜细胞凋亡的作用     

维甲酸在骨折愈合中的作用

目的研究外源性维甲酸(retinoicacid,RA)在骨折愈合中的作用.方法在骨折局部施加以自体冻干血凝块为载体的外源性RA,然后进行常规X线摄影和组织形态学观察.结果①X线术后第21d,实验组8例桡骨中有2侧可见缺损内完全为骨痂阴影充填,其余6侧也可见不同程度的阴影;对照组8侧只有少量的骨痂阴影充填.②组织学骨折第1、4d实验组与对照组相比,生发层总细胞数分别为(62.2±0.6)个/mm,(135.1±1.9)个/mm和(57.2±0.5)个/mm,(99.8±2.2)个/mm,两者间有显著差异(P＜0.01).骨折第4、7d实验组和对照组缺损区总细胞数分别为(63.7±1.4)个/mm,(55.0±0.2)个/mm和(45.1±1.1个)/mm,(51.9±0.7)个/nm,两者比较有显著差异和差异(P＜0.01和P＜0.05),第21d实验组和对照组成骨区体密度分别为(0.642±0.071)点/50um和(0.452±0.042)点/50um,两组差异显著(P＜0.01).结论RA具有加速骨折愈合的作用,并有望成为治疗骨折愈合的新方法.     

臀肌挛缩症的生物力学机制探讨

目的　探讨臀肌挛缩症患者临床表现的生物力学机制。方法　切取手术中得到的臀肌挛缩带及新鲜病尸的臀肌制成肌纤维束标本 ,在MTS生物力学测试机上测定其拉伸的力学性质。结果　臀肌挛缩带拉伸的弹性模量、极限强度和极限应变分别为 (2 .73 2± 0 .792 )N mm2 ,(1 48.3 2± 3 .84)MPa ,0 .896± 0 .3 1 5。而正常臀肌的弹性模量、极限强度和极限应变分别为 (0 .1 43± 0 .0 2 4)N mm2 ,(1 0 .5 0± 1 .69)MPa ,1 .43 4± 0 .40 2。两者间均有显著差异。结论　臀肌挛缩后 ,其强度和刚度增大 ,而弹性则减小 ,这是引起一系列临床表现的力学基础     

股骨颈轴向控制髓内钉与A—P型GAMMA钉的生物力学研究

目的研究股骨颈轴向控制髓内钉(axialcontrolledintramedullarynailofthefemoralneck,即ACINFN钉)的生物力学性能,为临床应用提供力学依据.方法24具新鲜尸体股骨标本随机分为对照组和两个实验组.对照组不作任何处理;实验组制成EvansⅠ型粗隆间骨折模型,分别用材料、规格相同的ACINFN钉和A-P型GAMMA钉固定.各组均在双轴液压伺服生物材料测试实验机上做抗压和抗扭转实验.结果在300N的压力下,对照组、ACINFN钉组和A-P型GAMMA钉组的压缩刚度分别为(1568.06±365.81)、(1166.53±91.8)和(224.11±66.45)N/mm,差异有显著性意义(P＜0.01);扭角为5°时,对照组、ACINFN钉组和A-P型GAMMA钉组的扭转刚度分别为(1.14±0.19)、(1.076±0.125)和(0.114±0.065)N@m/deg,差异有显著性意义(P＜0.01).结论ACINFN钉在抗压和抗扭转性能方面明显优于A-P型GAMMA钉,具有良好的生物力学性能.     

混合阳离子交联剂甲基丙烯酸海藻酸盐三维培养软骨细胞

本研究旨在建立一种更适合软骨细胞生长的三维培养体系。首先通过酰胺键反应将甲基丙烯酸引入海藻酸盐长链形成甲基丙烯酸海藻酸盐(MA),然后将MA和混合阳离子Ba2+∶Ca2+=5∶5进行光交联反应,形成内部结构为贯通式凝胶网络(IPN)的凝胶小球(MA凝胶小球)。将P1代软骨细胞包裹在MA凝胶小球中连续培养三周,在培养的不同时间点取适量凝胶小球进行倒置显微镜观察,冰冻切片后HE染色和免疫组化染色,扫描电镜观察第21d细胞在支架中的情况;通过Alamar blue法检测软骨细胞的增殖情况;二甲基亚甲兰法定量检测糖胺多糖(GAG)的含量。结果显示MA和混合阳离子Ba2+∶Ca2+=5∶5发生光交联反应形成的MA凝胶小球,与对照组藻酸钙相比,P1代软骨细胞在MA凝胶小球内增殖更明显,细胞外基质GAG的分泌也更多,扫描电镜下观察到软骨细胞可以良好地黏附在支架中生长,免疫组化染色呈阳性。以上结果证实软骨细胞在MA凝胶小球中的增殖和细胞外基质的分泌量优于藻酸钙组,并且能维持软骨细胞的表型,是一种更好的软骨细胞三维培养载体。     

视网膜细胞培养上清液对骨髓基质细胞的诱导分化作用

目的探讨视网膜细胞培养上清液对骨髓基质细胞(BMSCs)的诱导分化作用。方法原代培养胚龄17d(E17)及生后3d(P3)SD大鼠的视网膜细胞,分别收集培养上清液,过滤后与DMEM按2:3混合,用此混合液诱导BMSCs,倒置相差显微镜下观察细胞形态变化,对诱导后的细胞进行神经丝蛋白(NF)、胶质源性纤维酸性蛋白(GFAP)及视网膜特异性标记物进行免疫化学染色,统计分析阳性细胞数;再用RT-PCR进一步检测诱导结果。结果E17和P3细胞上清液均能诱导BMSCs表达NF[(10.35±2.66)%,(10.64±3.94)%]和GFAP[(27.86±7.97)%,(24.38±5.80)%],但只有P3细胞上清液诱导的BMSCs表达视蛋白(opsin),同时RT-PCR也检测到视细胞发育调节转录因子Crx mRNA的表达。结论视网膜细胞培养上清液可以诱导BMSCs向神经元、胶质细胞与视细胞分化。     

透骨消痛胶囊含药血清对软骨细胞外基质表达的影响**◆

背景：软骨细胞代谢异常引起细胞外基质降解和修复失衡是骨性关节炎的重要病理变化。透骨消痛胶囊具有补肾柔肝、活血祛风的功效,临床实验证实其对防治骨性关节炎有较好疗效。 目的：观察透骨消痛胶囊含药血清对软骨细胞外基质表达的影响。 方法：取SD大鼠膝关节软骨建立体外培养的软骨细胞,第3代软骨细胞同步化后进行干预。50只SD大鼠抽签法随机分为5组,用药量按照人体与动物药物等效剂量换算,空白组：生理盐水进行灌胃;对照组：壮骨关节丸水溶液灌胃;实验组：透骨消痛胶囊水溶液按0.145,0.290,0.580 mg/(g•d)进行灌胃。各组均连续给药3 d,采血前禁食12 h,末次给药1 h后腹主动脉采血,制备血清。 结果与结论：Ⅱ型胶原免疫组织化学染色可见透骨消痛胶囊实验组Ⅱ型胶原表达强于空白组及对照组;透骨消痛胶囊实验组CollagenⅡ、蛋白聚糖、Aggrecan mRNA基因表达明显升高(P < 0.01),Cathepsin K mRNA基因表达明显降低(P < 0.01)。提示透骨消痛胶囊含药血清能上调软骨细胞CollagenⅡ、蛋白聚糖、Aggrecan mRNA基因表达,下调Cathepsin K mRNA基因表达,促进细胞外基质合成,调控细胞外基质和软骨细胞之间的动态平衡。关键词：透骨消痛胶囊;骨性关节炎;含药血清;软骨细胞;细胞外基质 doi:10.3969/j.issn.1673-8225.2012.21.004     

不同浓度海藻酸盐凝胶的力学特性及其对软骨细胞增殖能力的影响

细胞外基质的理化性能影响着细胞的命运并调节组织形成。为探索最佳的软骨细胞体外三维生长环境,本研究通过改变藻酸盐浓度测定了海藻酸盐小球的力学特性及其对软骨细胞增殖能力的影响。凝胶的压缩机械性能通过INSTRON3365测定,结果发现浓度越高其压缩模量越大,表明凝胶对抗应变的能力越强。同时选取4w龄乳兔关节软骨细胞分别包裹在浓度为1%、2%和3%(w/v)海藻酸盐小球中连续培养4周,结果发现所有3组软骨细胞都表现出有限的增殖并形成类似于软骨组织的细胞簇,其中较低浓度组(1%和2%)增殖更明显。这些结果表明软骨细胞在海藻酸盐凝胶小球中连续培养有少量增殖,且较低浓度的藻酸盐更有利于软骨细胞的增殖,说明藻酸盐凝胶的力学特性影响了软骨细胞的增殖能力。     

CPT-cAMP对成年金黄地鼠视网膜节细胞存活的影响

目的　探讨眼球玻璃体内注射CPT cAMP对切断视神经后视网膜节细胞存活的影响。方法　用荧光素逆行示踪标记法和定量解剖学技术观察视神经切断 5、7和 14d后成年金黄地鼠视网膜节细胞的密度。结果　 (1)正常视网膜节细胞平均密度为 2 0 0 7± 115 /mm2 ;(2 )视神经切断 5、7、14d后 ,视网膜节细胞平均密度分别下降至 :10 10± 131/mm2 、782± 5 5 /mm2 和2 14± 30 /mm2 ;(3)给予DMSO/生理盐水的对照组在上述各时间段视网膜节细胞平均密度与单纯神经切断组结果相似 ;(4)给予CPT cAMP的实验组在 5、7、14d视网膜节细胞的平均密度分别为 1398± 2 45 /mm2 、12 93± 84/mm2 和 5 0 1± 72 /mm2 ,与视神经切断组和DMSO/生理盐水对照组相比在各时间段上均存在显著性差异 (P <0 0 5 )。结论　CPT cAMP可提高视神经切断后成年金黄地鼠视网膜节细胞的存活。     

抗原致敏的DC-CIK对前列腺癌细胞DU145杀伤的作用

目的探讨抗原致敏的DC与CIK细胞共同培养后获得的DC-CIK细胞(Ag-DC-CIK)对前列腺癌细胞株DU145的杀伤作用。方法分离健康志愿者外周血单个核细胞,贴壁细胞以GM-CSF、IL-4及TNF-α诱导并加入DU145全细胞冻融抗原培养抗原致敏的DC,非贴壁细胞以IL-2、IL-1α、IFN-γ和mAb诱导培养CIK细胞,并将两种细胞共同培养6d获得Ag-DC-CIK;每隔2天计数细胞;流式细胞术检测细胞免疫表型;以共培养6d的Ag-DC-CIK作为效应细胞,以相同天数未致敏的DC-CIK和CIK细胞为对照组,DU145细胞作为靶细胞,采用MTT法分析各组杀瘤率。结果外周血单核细胞成功诱导DC,Ag-DC-CIK细胞培养6d的扩增倍数为CIK的1.69-2.56倍(P<0.05)。CIK组、DC-CIK组及Ag-DC-CIK组中CD3+CD56+细胞所占百分比分别为(19.15±1.55)%,(28.43±1.51)%和(39.12±2.29)%,依次升高(P<0.01),对前列腺癌DU-145细胞24h杀瘤率分别为(26.39±4.47)%,(46.82±5.68)%,(62.80±2.01)%,依次升高(P<0.01)。结论 Ag-DC-CIK细胞增殖活性强,对前列腺癌DU-145细胞有明显杀伤作用,强于未致敏DC-CIK细胞和CIK细胞。     

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 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.     

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.     

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.     

Environmental risk factors and their role in the management of atopic dermatitis

Introduction: The etiology of atopic dermatitis (AD) is multifactorial with interaction between genetics, immune and environmental factors.

Areas covered: We review the role of prenatal exposures, irritants and pruritogens, pathogens, climate factors, including temperature, humidity, ultraviolet radiation, outdoor and indoor air pollutants, tobacco smoke exposure, water hardness, urban vs. rural living, diet, breastfeeding, probiotics and prebiotics on AD.

Expert commentary: The increased global prevalence of AD cannot be attributed to genetics alone, suggesting that evolving environmental exposures may trigger and/or flare disease in predisposed individuals. There is a complex interplay between different environmental factors, including individual use of personal care products and exposure to climate, pollution, food and other exogenous factors. Understanding these complex risk factors is crucial to developing targeted interventions to prevent the disease in millions. Moreover, patients require counseling on optimal regimens for minimization of exposure to irritants and pruritogens and other harmful exposures.     

'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 genes in Amerindians from Mexico San Vicente Tancuayalab Teenek/Huastecos

Huastecos or Teenek Amerindians are presently living at North East Mexico (San Luis Potosi State). They have probably one of the most ancient culture of Mexico and Central America together with Mayas and Olmec groups with which also show close relationships. Proximity to Atlantic Ocean/Mexican Gulf originated that Spaniards had very early contact with them at about 1519 CE or before. In the present paper we have aimed to study HLA gene profile which may be useful for HLA and disease epidemiology and transplant programs in Teeneks. HLA-DRB1*04:07, -DRB1*14:06 and -DRB1*04:11 have been found in high frequency like in other Amerindian groups. High frequency typical Amerindians HLA extended haplotypes have been found, such as A*02-B*35-DRB1*04:07-DQB1*03:02; A*68-B*39-DRB1*04:07-DQB1*03:02 and A*02-B*39-DRB1*04:07-DQB1*03:02; also new haplotypes have been described, like A*02-B*52-DRB1*04:11-DQB1*03:02, A*68-B*35-DRB1*14:02-DQB1*03:01 and A*68-B*40-DRB1*16:02-DQB1*03:01. Genetic proximity is observed not only to linguistically close Mayans, but also to Mazatecans, Mixtecans and Zapotecans, who speak an altogether different languages; it shows once more that genes and languages do not correlate. This population was greatly diminished after European contact between 1500 and 1600 years CE; in fact, North and South America First Inhabitants population was brought from 80 down to 8 million people because of diseases (i.e.: measles, smallpox or influenza), slavery and war.     

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.