温阳益气平喘方对支气管哮喘大鼠嗜酸粒细胞凋亡和相关基因蛋白Bcl-2和Bax表达的影响

目的:研究温阳益气平喘方对支气管哮喘大鼠肺组织嗜酸粒细胞凋亡及相关基因蛋白Bcl-2和Bax表达的影响.方法:60只SD大鼠随机分为6组,即正常组、模型组、温阳益气平喘高剂量组、温阳益气平喘低剂量组、桂龙咳喘宁组,氨茶碱组,每组10只.除正常组外以卵蛋白致敏并吸入激发法制备大鼠支气管哮喘模型,各治疗组均从第1次哮喘激发开始(造模第3周)至处死前每天灌胃给药,激发并给药4周后处死大鼠.应用TUNEL法检测肺组织嗜酸粒细胞凋亡,免疫组织化学检测肺组织Bcl-2和Bax蛋白的表达.结果:温阳益气平喘方高低剂量组大鼠肺组织嗜酸粒细胞凋亡指数显著高于哮喘模型组(P＜0.01和P＜0.05),高低剂量组优于桂龙咳喘宁组(P＜0.01),高剂昔组优于氨茶碱组(P＜0.01);肺组织及气道壁中Bcl-2蛋白阳性细胞的比例显著低于哮喘模型组(P＜0.01和P＜0.05),高剂量组优于桂龙咳喘宁和氨茶碱组(P＜0.01);Bax蛋白阳性细胞的比例显著高于哮喘模型组(P＜0.01),高低剂量组优于桂龙咳喘宁组(P＜0.01),高低剂量组优于氨茶碱组(P＜0.01和P＜0.05).结论:温阳益气平喘方可通过调节抑制凋亡基因bcl-2和促进凋亡基因bax蛋白的表达,促进肺内EOS凋亡,减少炎细胞浸润,减轻哮喘气道炎症.     

慢性乙型肝炎PBMC凋亡及淋巴细胞亚群的检测

目的了解慢性/慢性重型乙型肝炎外周血淋巴细胞激活诱导细胞死亡(AICD)现象的存在情况、各免疫细胞的状况、AICD与淋巴细胞状况的关系,以探讨乙型肝炎慢性化和重型化的机制.方法利用慢性、慢性重型乙型肝炎病人和健康献血员外周血单个核细胞(PBMC)在PHA-P刺激下培养72h,通过流式细胞仪检测PBMC的凋亡情况;采用流式细胞仪结合全自动血液分析仪对慢性、慢性重型乙型肝炎病人及正常对照组外周血各淋巴细胞亚群进行检测.结果慢性乙型肝炎组PBMC凋亡率高于慢性重型乙型肝炎组(P＜0.01),高于正常对照组(P＜0.01).慢性乙型肝炎组总淋巴细胞百分率高于慢性重型乙型肝炎组(P＜0.01),慢性重型乙型肝炎组淋巴细胞数低于正常对照组(P＜0.01);CD3+、CD3+CD4+细胞数低于正常对照组(P＜0.01),低于慢性乙型肝炎组(P＜0.05);CD3+CD8+细胞数低于正常对照组(P＜0.05),低于慢性乙型肝炎组(P＜0.05).单核细胞百分率高于正常对照组(P＜0.01),高于慢性乙型肝炎组(P＜0.05).结论慢性乙型肝炎患者外周血淋巴细胞活化与凋亡共存,慢性重型乙型肝炎患者外周血淋巴细胞消耗严重,AICD参与乙型肝炎慢性化、重型化的发生机理.     

甘肃党参水煎剂对D-半乳糖诱导衰老小鼠免疫功能的影响

目的 研究甘肃党参水煎剂对D-半乳糖诱导衰老小鼠免疫功能的影响.方法 采用D-半乳糖诱导亚急性衰老小鼠模型,测定每组小鼠的体质量,计算脾脏指数和胸腺指数,观察脾脏组织超微结构,采用免疫组化法检测CD138的表达,ELISA检测小鼠血清中IgG、IgM和补体C3、C4含量.结果 与正常对照组相比,衰老模型组小鼠的脾脏指数和胸腺指数显著降低(P＜0.01),与模型组比较,高、低剂量党参组和维生素E组小鼠的脾脏指数和胸腺指数显著升高(P＜o.ol).正常对照组脾小体中淋巴细胞的超微结构均正常,而模型组小鼠的脾脏超微结构有明显的损伤现象.低、高剂量党参组和维生素E组均使已受损伤的脾脏超微结构得以修复,其中高剂量党参组使受损的脾脏超微结构恢复到最接近正常水平.与正常对照组比较,衰老模型组小鼠脾脏组织中CD138的表达率较低(P＜0.01),小鼠血清中IgG、IgM和补体C3、C4含量显著减少(P＜0.01或P ＜0.05);与衰老模型组比较,正常对照组和高剂量党参组小鼠脾脏组织中CD138的表达率较高(P＜0.01),小鼠血清中IgG、IgM和补体C3、C4含量显著增加(P＜0.01或P＜0.05),而党参低剂量组和维生素E组无显著差异(P＞0.05).结论 甘肃党参水煎剂具有增强D-半乳糖诱导衰老小鼠免疫功能的作用.     

卡维地洛对病毒性心肌炎心肌细胞凋亡及慢性期胶原重塑的干预作用及其机制的实验研究

目的 探讨卡维地洛对病毒性心肌炎心肌细胞凋亡及慢性期胶原重塑的干预作用及其机制.方法 将140只雄性BALb/c小鼠随机分为模型组(60只)、治疗组(60只)和对照组(20只).模型组和治疗组小鼠给予腹腔注射CVB3,建立模型.治疗组于注射病毒1h后给予卡维地洛1.0 mg/kg/d.在接种21d后检测基质金属蛋白酶(MCP-1),肿瘤坏死因子α(TNF-α),一氧化氮合酶(iNOS),核因子κB(NF-κB)表达水平以及心肌细胞间质胶原容积分数(CVF)和细胞凋亡率(Rapo),测量各组小鼠的血压和心脏的质量及体质量;对比分析各组小鼠的心脏组织切片.结果 21d后模型组存活33只,治疗组存活44只,对照组全部存活,三组间具有统计学差异(x2=15.082,P＜0.01).模型组和治疗组小鼠MCP-1,TNF-α,iNOS,NF-κB表达水平显著高于对照组(P＜0.01),且治疗组显著低于模型组(P＜0.01).治疗后心肌细胞凋亡率和CVF显著高于对照组(P＜0.01),且治疗组显著低于模型组(P＜0.01).结论 卡维地洛可以有效降低病毒性心肌炎的炎性因子并以此达到保护心肌细胞、促进胶原重塑的作用.     

胎盘部位滋养细胞肿瘤中NF-κBp65、CD44v6、Kiss-1的表达及相关性

目的 探讨细胞核因子-κBp65(NF-κBp65)、CD44v6、Kiss-1在胎盘部位滋养细胞肿瘤中的表达及相关性.方法 采用免疫组化PV 6000两步法检测20例正常早孕绒毛、15例葡萄胎、15例绒癌、10例胎盘部位滋养细胞肿瘤(placent site trophoblastic tumor,PSTT)中NF-κBp65、CD44v6、Kiss-1的表达情况及相关性.结果 NF-κBp65、CD44v6分别在正常早孕绒毛、葡萄胎、PSTT、绒癌中表达呈升高趋势,差异有统计学意义(P1＜0.01、P2＜0.01).NF-κBp65、CD44v6在PSTT中表达高于正常早孕绒毛和葡萄胎中的表达,差异有统计学意义(P1＜0.01、P2＜0.01、P3＜0.01、P4＜0.05);NF-κBp65在PSTT和绒癌中表达差异有统计学意义(P＜0.01);CD44v6在PSTT和绒癌中表达差异无统计学意义(P>0.05).Kiss-1在正常早孕绒毛、葡萄胎、PSTT、绒癌中表达呈下降趋势,差异有统计学意义(P＜0.01).Kiss-1在PSTT中表达低于正常早孕绒毛和葡萄胎中的表达,差异有统计学意义(P1＜0.01,P2＜0.01);在PSTT和绒癌中表达差异无统计学意义(P>0.05).NF-κBp65和CD44v6呈正相关(r=0.356,P=0.005)和Kiss-1呈负相关(r=-0.527,P=0.000);CD44v6和Kiss-1呈负相关(r=-0.346,P=0.007).结论 NF-κBp65、CD44v6的高表达和Kiss-1的低表达可能在胎盘部位滋养细胞肿瘤的浸润转移中起重要作用.     

脑室注射链脲佐菌素对大鼠海马神经元凋亡相关蛋白表达的影响

目的 观察App17肽对脑室注射链脲佐菌素的大鼠海马神经元凋亡相关蛋白表达的影响,探讨胰岛素信号转导通路障碍对神经元存活的作用.方法脑室注射链脲佐菌素(STZ)制作大鼠痴呆模型.3周后,皮下注射App17肽.4周后取脑组织做Bcl-2、Bax、CytoC免疫组织化学染色及Western blotting半定量分析.结果 模型组大鼠海马内Bax、CytoC阳性反应神经元细胞数目多,胞质深染,细胞计数与正常组及治疗组有显著性差异(P＜0.01);模型组大鼠海马内Bcl-2阳性细胞数目少,胞质染色淡,细胞计数与正常组及治疗组有显著性差异(P＜0.01).Western blotting半定量分析可见、Bcl-2、Bax、CytoC出现清晰的条带,组间可见较明显的差异(P＜0.01).结论 脑室注射STZ的大鼠海马内促进凋亡的Bax、CytoC表达增加;抑制凋亡的Bcl-2表达降低.App17肽可影响上述蛋白的表达,使之接近正常.胰岛素信号转导通路对神经元存活有一定作用.     

糖尿病大鼠胃平滑肌细胞凋亡与线粒体膜电位改变

目的 揭示糖尿病胃轻瘫(DGP)发病机制 方法 SD大鼠随机分为对照组和模型组,造模10周后,模型检测;流式细胞术测细胞凋亡与 m;免疫组化测CYT C。结果 与对照组相比：(1)模型组造模后均出现多尿、多饮、多食症状,体重减轻;血糖浓度显著增高 (P＜0.01);胃内色素残留率显著降低（P＜0.01）。(2)模型组胃平滑肌细胞凋亡率显著增高（P＜0.01）, m显著降低（P＜0.01）, 胞质CYT C显著增加。结论 线粒体介导了糖尿病大鼠胃平滑肌细胞凋亡,后者在糖尿病胃轻瘫发病机制中扮演了一定角色。     

揉髌手法对兔膝关节软骨细胞凋亡及Bcl-2、Bax和Fas表达的影响

背景：现代研究证实细胞的过度凋亡加速软骨组织的退变,而手法的力学刺激对关节软骨影响的分子层面研究至今少有报道。 目的：观察揉髌手法对兔膝关节软骨细胞凋亡及Bcl-2、Bax、Fas表达的作用。 方法：50只新西兰兔随机等分为正常组、假手术组、模型组、手法组和针剂组,后3组建立右下肢骨内高压型膝关节骨性关节炎模型。造模1周后,手法组使用揉髌手法隔天治疗1次,每次10 min,共治疗5周共17次;针剂组关节内注射玻璃酸钠液0.6 mL,每周1次,共5次。 结果与结论：造模后8周末取兔右膝关节内侧胫骨平台软骨组织,苏木精-伊红染色提示模型组软骨组织退变明显,软骨细胞凋亡率明显增加(P < 0.01),胫骨平台软骨组织中Bcl-2、Bax、Fas表达率明显升高(P < 0.01),而手法组和针剂组软骨组织退变轻微,软骨细胞凋亡率及Bax、Fas表达率较模型组降低(P < 0.01),但胫骨平台软骨组织中Bcl-2表达升高(P < 0.01)。说明揉髌手法与关节内注射玻璃酸钠一样可以明显降低兔膝关节软骨细胞的凋亡率,其作用机制与上调Bcl-2表达,下调Bax、Fas表达有关。     

新生大鼠缺氧缺血性脑损伤对脾bcl-2/bax/fas/fasL mRNA的影响

目的 通过检测脾淋巴细胞凋亡基因bcl-2/bax/fas/fasL mRNA表达水平,探讨缺氧缺血性肭损伤(hypoxicischemic brain damage,HIBD)脾细胞凋亡基因的变化与缺氧时间的关系,以此提供HIBD对于免疫功能影响的理论基础.方法 建立新生大鼠HIBD模型测定脾bcl-2/bax/fas/fasL mRNA 的相对表达量.结果 bcl-2处理组相对表达量低于对照组(P＜0.01).bax处理组相对表达量高于对照组(P＜0.01).bcl-2/bax处理组比值低于相应对照组(P＜0.05).fas处理纽相对表达量高于对照组(P＜0.01).fasL 1 h和6 h处理组相对表达量高于对照组,12 h之后则对照组远高于处理组.fas/fasL对照组较分散,除6 h组外其余组均小于1,处理组较集中,除24 h外均大于1.结论 HIBD可促进脾细胞促凋亡基因的表达,减少抑凋亡基因表达;HIBD发生6～12 h是促凋亡基因和抑凋亡基因高峰表达期;HIBD加强了fas/fasL的作用.     

自然流产模型小鼠蜕膜细胞凋亡及相关基因的表达

目的 通过比较正常妊娠模型小鼠及自然流产模型小鼠蜕膜细胞凋亡及Bcl-2、Bax、Fas、FasL蛋白的表达,从细胞及分子水平探讨自然流产的发病机制.方法建立正常妊娠模型CBAXBALB/c和自然流产模型CBAXDBA/2.用免疫组化SABC法测定两组模型孕13 d蜕膜细胞Bcl-2、Bax、Fas、FasL蛋白的表达,并通过MIAS-2000医用彩色病理图像免疫组化测量系统对其表达进行半定量分析,其结果用平均灰度值表示;同时应用DNA缺口原位末端标记技术(TUNEL)测定两组模型孕13天蜕膜细胞凋亡情况.结果与正常妊娠模型相比,自然流产模型蜕膜细胞Bcl-2蛋白的表达降低(P＜0.01);Bax蛋白的表达明显升高(P＜0.01);FasL的表达明显升高(P＜0.01);Fas的表达两组比较无明显差异(P＞0.05).蜕膜细胞凋亡指数(AI),自然流产模型明显高于正常妊娠模型(P＜0.01).结论 早孕期蜕膜组织细胞凋亡异常是自然流产的机制之一,Bcl-2/Bax,Fas/FasL途径可能是诱导早孕期蜕膜细胞凋亡的重要因素.     

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.