基于垂体腺苷酸环化酶激活多肽在帕金森动物模型中的神经保护作用及机制探索

目的探讨垂体腺苷酸环化酶激活多肽 （PACAP） 在帕金森病 （PD） 动物模型中的神经保护作用及机制。方法 SD大鼠设置假手术组 （Sham组）、PD组 （帕金森造模组）、PD+PACAP_低浓度组 （造模+按体重 5 μg/kg予 PACAP鼻腔给药）、PD+PACAP_中浓度组 （造模+按体重10 μg/kg予PACAP鼻腔给药）、PD+PACAP_高浓度组 （造模+按体重20 μg/kg予 PACAP鼻腔给药） 5组,最终每组纳入10只。四个PD组将脂多糖溶液注射至SD大鼠黑质中建立PD模型,假手术组同法操作但注射生理盐水。通过转棒法检测大鼠行为学表现;ELISA 法检测黑质谷氨酸浓度 （Glu） 和 α-突触核蛋白 （α-syn） 水平,WB法检测黑质α-syn及核因子激活的B细胞的κ-轻链 （NF-κB）、核因子κB抑制蛋白α （IkBα） 的表达水平;实时荧光定量聚合酶链式反应法 （qPCR） 检测大鼠黑质炎症因子肿瘤坏死因子 α （TNF-α）、白细胞介素-1β （IL-1β）、白细胞介素6IL-6的mRNA表达水平;WB法和免疫荧光法检测核转录因子胶质纤维酸性蛋白 （GFAP） 水平来观察星形胶质细胞的活化水平,TUNEL法检测黑质神经元的凋亡情况。结果 （1） 行为学结果显示,在转棒实验中,与Sham组比较,PD组潜伏期缩短,掉落次数增加 （37.20±5.27 次/min）;与 PD 组比较,PACAP 组潜伏期延长,掉落次数减少 （14.50±2.32 次/min）, PACAP 治疗组的潜伏期和掉落次数随着 PACAP 浓度的增加而潜伏期可以更长,掉落次数可以更少 （P＜0.001）。（2） 星形胶质细胞激活相关指标显示,PD组 GFAP表达水平和黑质神经元的凋亡数量较 Sham组显著升高;PACAP治疗组上述指标随着 PACAP的浓度增加而显著减低 （P＜0.01）。（3） 黑质神经炎症指标显示,与 Sham 组比较,PD组 NF-κB含量及TNF-α、 IL-1β、IL-6的mRNA显著升高,IkBα含量显著降低;PACAP治疗组NF-κB含量及TNF-α、IL-1β、IL-6的mRNA随着PACAP 的浓度增加而减低,IkBα含量随着PACAP的浓度增加而升高（P＜0.01）。（4）PD疾病特征指标显示,PD组谷氨酸和α-syn含量均较Sham组显著升高,3个PACAP组的上述指标较PD组显著降低;PACAP治疗组的上述指标随着PACAP的浓度增加而递减 （P＜0.01）,以 PD+PACAP_高浓度组效果最为明显。结论 PACAP在 PD模型中具有神经保护作用,其机制可能为通过抑制星形胶质细胞激活来减轻神经炎症的发生。     

外固定支架术后行内固定治疗对胫腓骨骨干开放性骨折患者的疗效分析

目的 探究外固定支架术后行内固定治疗对胫腓骨骨干开放性骨折患者的影响。方法 选取2018年1月至2021年1月宁德师范学院附属宁德市医院收治的87例胫腓骨骨干开放性骨折患者为研究对象，根据手术方式分为A组（42例）和B组（45例），A组采用外固定支架术治疗，B组采用外固定支架术后行内固定治疗。比较两组患者的骨折愈合情况、下肢功能及自理能力恢复情况、骨代谢标志物含量、炎症反应及并发症发生率。结果 患者均获随访6 ～ 8个月。B组患者的骨痂形成时间、疼痛消失时间、骨折愈合时间均少于A组，术后6个月B组骨折愈合率高于A组（P＜0.05）；术后3个月及术后6个月，B组患者的膝踝关节功能恢复评分（Merchant）、简式Fugl-Meyer（FMA）下肢运动量表评分、生活自理能力恢复有效性（BE）值均高于A组（P＜0.05）。术后1周，B组患者的骨碱性磷酸酶（BALP）、骨钙素（BGP）、血清I型前胶原羧基端肽（PICP）含量均高于A组（P＜0.05）；B组患者的C反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）水平均低于A组（P＜0.05）；B组患者的并发症总发生率低于A组（P＜0.05）。结论 采用外固定支架术后行内固定治疗胫腓骨骨干开放性骨折安全有效，能够缩短骨折端与创面的愈合进程，促进患者功能恢复，减少并发症的发生，改善预后。     

党参多糖调控miR-361-5p/TLR4/NF-κB通路对胃癌细胞AGS增殖、凋亡和炎症因子表达的影响

目的 探讨党参多糖对胃癌细胞AGS增殖、凋亡和炎症因子表达的影响和机制。方法 采用CCK-8实验、流式细胞术评估党参多糖（10、20、40μmol/L）对胃癌细胞AGS活力和凋亡的影响。RT-qPCR检测miR-361-5p表达;ELISA法检测肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-1β分泌。蛋白质免疫印记法检测TLR4和磷酸化核因子（NF）-κBp65(p-p65)和磷酸化NF-κB抑制蛋白-α(p-IκBα)蛋白的表达。将miR-361-5p模拟物、miR-361-5p抑制物分别转染AGS细胞,经40μmol/L党参多糖处理后,用上述方法检测AGS细胞活力、凋亡率、Toll样受体4(TLR4)、p-p65和p-IκBα蛋白表达以及TNF-α、IL-6、IL-1β分泌。结果 10、20、40μmol/L党参多糖显著降低AGS细胞活力（P<0.05）,增加细胞凋亡率（P<0.05）,抑制TNF-α、IL-6、IL-1β分泌（P<0.05）,并上调miR-361-5p表达水平（P<0.05）。40μmol/L党参多糖明显降低TLR4、p-p65...     

采用“双侧垂直错位穿刺法”与“双侧水平裂隙穿刺法”行经皮椎体成形术在治疗Ⅰ期Kümmell病的疗效对比

目的 探究采用“双侧垂直错位穿刺法”与“双侧水平裂隙穿刺法”行经皮椎体成形术（PVP）在治疗I期Kümmell病中的疗效对比。方法 回顾性分析2014年1月至2021年1月在中国人民解放军火箭军特色医学中心接受PVP治疗的I期Kümmell病患者53例，其中采取“双侧垂直错位穿刺法”治疗25例（A组），采取“双侧水平裂隙穿刺法”治疗28例（B组）。比较两组的手术时间、骨水泥注入量、骨水泥渗漏率、骨水泥分布及并发症情况，并记录术前与术后1 d、1个月、3个月、6个月视觉评分量表（VAS）和Oswestry功能障碍指数（ODI），通过X线或CT检查测量并记录术前、术后伤椎前缘与中线高度和Cobb角，并观察骨水泥松动或移位及伤椎再骨折发生情况，末次随访采用改良MacNab标准评估手术的优良率。结果 两组患者均顺利完成手术，均获定期随访。A组较B组手术时间相当（P＞0.05），骨水泥注入量更多（P＜0.05）；A组术中2例出现骨水泥渗漏，渗入椎间盘1例、椎体前侧１例，无临床症状；B组术中9例出现骨水泥渗漏，渗入椎间盘6例、椎体前侧3例，无临床症状；A组术中骨水泥渗漏率较低（P＜0.05），术后椎体裂隙及骨折线周围骨水泥分布更广泛（P＜0.05）；两组术中均无脊髓、椎前大血管损伤，无肺栓塞等并发症发生。与术前比较，两组患者术后1 d、1个月、3个月、6个月VAS、ODI显著降低（P＜0.05）；A组与B组比较，术前、术后VAS、ODI在两组间差异无统计学意义（P＞0.05）；与术前比较，两组患者术后1 d、1个月、3个月、6个月伤椎前缘与中线高度显著增加（P＜0.05），Cobb角测量值显著降低（P＜0.05）；A组与B组比较，术后1 d、1个月、3个月、6个月伤椎前缘与中线高度变化、伤椎Cobb角变化差异无统计学意义（P＞0.05）；末次随访时两组患者均无骨水泥松动或移位及伤椎再骨折情况发生，采用改良MacNab标准评定临床疗效：A组优23例、良2例，B组优19例、良9例，A组优秀率高于B组（P＜0.05）。结论 治疗Ⅰ型Kümmell病时，与“双侧水平裂隙穿刺法”经皮椎体成形术相比，“双侧垂直错位穿刺法”经皮椎体成形术术中的骨水泥渗漏率更低，骨水泥分布更广泛，术后早期治疗效果相当，中期效果更确切。     

藤黄健骨胶囊通过SIRT1/NF-κB/NLRP3信号通路调节绝经后骨质疏松大鼠破骨细胞分化

目的：探讨藤黄健骨胶囊（TJC）对绝经后骨质疏松大鼠沉默信息调节因子1(SIRT1)/核因子κB(NF-κB)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)信号通路关键蛋白表达的影响及分子机制。方法：采用随机数字表法将SD雌性大鼠分为假手术组，模型组[卵巢切除术（OVX）组]，SIRT1抑制剂组（EX-527+OVX组），阳性药物戊酸雌二醇（ESV）对照组（ESV+OVX组），以及高、中、低剂量TJC处理组（TJC-H+OVX、TJC-M+OVX和TJCL+OVX组），每组10只。OVX建模，建模成功后给予TJC或ESV干预，并根据实验设计给予EX-527干预，8周后取材。ELISA法检测相关炎症因子水平；双重免疫荧光染色分别检测SIRT1、NF-κB p65和NLRP3蛋白；RT-qPCR和Western blot分别检测SIRT1/NF-κB/NLRP3信号通路相关分子和破骨分化标志分子的mRNA和蛋白水平。结果：与假手术组相比，OVX组和EX-527+OVX组大鼠血清中白细胞介素1β(IL-1β)、IL-18和肿瘤坏死因子α(TNF-α)含量显著升高（P<0.01），破...     

急性热应激对睾丸免疫微环境相关蛋白的影响

目的：研究急性热应激对睾丸免疫微环境相关蛋白的影响。方法：49只雄性SD大鼠随机分为对照组和热应激组（0.5 h组、2 h组、4 h组、6 h组、24 h组和48 h组）。热应激组大鼠麻醉后43℃恒温水浴20 min,并分别于0.5 h、2 h、4 h、6 h、24 h和48 h后摘取睾丸。Western blot检测睾丸内occludin、ZO-1、vimentin、p-NF-κB p65和p-IκBα的表达;ELISA法检测睾丸内IL-1β表达。结果：与对照组相比,热应激组vimentin和occludin表达均显著降低（P<0.01）;0.5 h组ZO-1表达差异无统计学意义（P>0.05）,其余热应激组ZO-1表达显著降低（P<0.01）;0.5 h组、2 h组和4 h组睾丸内p-NF-κB p65和p-IκBα表达与对照组相比差异无统计学意义（P>0.05）,6 h组、24 h组和48 h组p-NF-κB p65和p-IκBα表达显著升高（P<0.01）。与对照组相比,24 h组和48 h组睾丸内IL-1β表达差异无统计学意义（P>0.05...     

富血小板血浆复合自体松质骨移植的生物重建治疗陈旧性腕舟状骨骨不连的临床效果分析

目的 观察富血小板血浆（platelet-rich plasma, PRP）复合自体松质骨移植治疗腕舟状骨骨不连的近期临床效果。方法 收集中国人民解放军联勤保障部队第989医院自2019年6月至2021年6月期间收治的30例陈旧性腕舟状骨骨不连的患者作为研究对象,按照随机数表分为观察组和对照组。观察组（15例）采用PRP复合自体松质骨移植的生物重建方法,对照组（15例）采用只采用自体松质骨移植并内固定的手术方式。对比两组的手术时间、住院时间、骨折愈合情况、骨痂等级评分、腕关节功能评估及术后并发症。结果 术后30例患者均得到随访,随访周期为12个月。观察组骨折愈合率为93.3%,对照组骨折愈合率为66.7%,两组比较差异具有统计学意义（P＜0.05）。观察组的手术时间为（1.83±0.23）h,对照组的手术时间为（1.73±0.30）h,两组比较差异无统计学意义（P＞0.05）;观察组骨折愈合时间［（16.54±0.74）周］少于对照组［（20.70±4.73）周］,术后住院时间［（14.27±2.43） d］少于对照组［（19.53±6.29） d］,两组比较差异具有统计学意义（P＜0.05）。观察组术后4、8、12周骨痂等级评分比较优于对照组,两组比较差异具有统计学意义（P＜0.05）;观察组术后16周骨痂等级评分比较优于对照组,两组比较差异无统计学意义（P＞0.05）。数据显示术后6个月,观察组的Mayo评分高于对照组,VAS评分低于对照组,两组比较差异具有统计学意义（P＜0.05）;术后12个月,两组间Mayo评分及VAS评分比较,差异无统计学意义（P＞0.05）。观察组术后并发症共2例,对照组术后并发症共6例,两组比较差异具有统计学意义（P＜0.05）。结论 富血小板血浆（PRP）复合自体松质骨移植治疗腕舟状骨骨不连可促进腕舟状骨骨折愈合,减少手术相关并发症,利于腕关节功能恢复,近期临床效果较好。     

闭合撬拨复位经皮螺钉内固定术与斜置塑形钢板内固定术在舌形跟骨骨折中的疗效对比

目的 探讨采取闭合撬拨复位经皮螺钉内固定术与斜置塑形钢板内固定术治疗舌形跟骨骨折的疗效对比。方法 回顾性选取四川省骨科医院2016年1月至2018年5月收治的舌形跟骨骨折患者68例，按手术方法的不同将患者分为两组，其中采用闭合撬拨复位经皮螺钉内固定术治疗的35例患者（闭合复位组），余下33例采用斜置塑形钢板内固定治疗（斜置钢板组），观察两组的手术时间、住院时间、骨折愈合时间及术后跟骨高度、Böhler角、并发症和Maryland足踝功能评分。结果 68例患者获得术后3年随访，闭合复位组患者的手术时间、住院时间、骨折愈合时间明显短于斜置钢板组（P＜0.05）；术后两组患者跟骨高度、Böhler角比较，差异无统计学意义（P＞0.05）；两组患者并发症总发生率及Maryland足踝功能评分比较，差异无统计学意义（P＞0.05）。结论 闭合撬拨复位经皮螺钉内固定治疗舌形跟骨骨折能缩短手术时间、住院时间和骨折愈合时间，与斜置塑形钢板内固定相比，在纠正跟骨的高度、Böhler角和并发症方面差异不大。     

切开复位钢板内固定与闭合复位经皮螺钉固定治疗Sanders Ⅱ、Ⅲ型跟骨骨折疗效比较

目的 分析比较切开复位钢板内固定与闭合复位经皮螺钉固定治疗Sanders Ⅱ、Ⅲ型跟骨骨折的临床疗效。方法 回顾性分析2009年1月至2019年10月鄂东医疗集团黄石市中医医院收治的Sanders Ⅱ、Ⅲ型跟骨骨折患者84例（88足）,其中采用切开复位钢板内固定的（切开复位组）有41例（42足）,采用闭合复位经皮螺钉固定的（闭合复位组）有43例（46足）。比较两组患者的受伤至手术时间、手术时间、术中出血量、手术切口并发症及跟骨解剖参数,并采用美国足踝外科协会（AOFAS）评分系统进行踝关节功能评估。结果 闭合复位组的受伤至手术时间、手术时间及术中出血量明显优于切开复位组,闭合复位组手术切口并发症也要明显少于切开复位组,差异有统计学意义（P＜0.05）。跟骨解剖参数恢复程度上,跟骨长度、宽度及高度比较：术前两组比较,差异无统计学意义（P＞0.05）;两组患者术前与术后比较,差异有统计学意义（P＜0.05）;但是术后两组比较,差异无统计学意义（P＞0.05）。跟骨Böhler及Gissane角比较：术前两组比较,差异无统计学意义（P＞0.05）;两组患者术前与术后比较,差异有统计学意义（P＜0.05）;术后切开复位组在跟骨Böhler角及Gissane角恢复的情况上优于闭合复位组,差异有统计学意义（P＜0.05）。踝关节功能评分闭合复位组的优良率为73.9%,切开复位组为76.2%,两组比较差异无统计学意义（P＞0.05）。结论 在确保临床疗效情况下,闭合复位经皮螺钉固定在治疗Sanders Ⅱ及部分Sanders Ⅲ型跟骨骨折上有着受伤至手术时间短、手术时间短、术中出血量少及并发症低的优点,是一种很好的微创治疗方法。     

新型腹腔降温法对心肺复苏后兔炎症反应的影响

目的:观察心肺复苏(CPR)后不同降温方法对自主循环恢复(ROSC)后兔炎症反应的影响。方法:48只新西兰成年大白兔,采用交流电致颤的方式建立CPR模型, 依据降温方式的不同,随机分为常温组(NT)、腹腔内降温组(PC)、体表降温组(SC)和头部局部降温组(LC)。观察不同降温方法对ROSC后血浆TNF-α和IL-6浓度的影响,ROSC后12 h取肝脏组织,Western blotting检测NF-κB p65和NF-κB p50蛋白表达。记录每组动物的存活时间并进行比较。结果:ROSC后4 h、8 h和12 h,PC组血浆TNF-α水平显著低于NT组;至12 h时PC组也显著低于SC组和LC组(P<0.05);ROSC后2 h、4 h、8 h和12 h,PC组血浆IL-6水平明显低于NT组(P<0.05),而与SC、LC组间无显著差异。ROSC后12 h,PC组肝核蛋白内NF-κB p65和p50蛋白表达水平显著低于其它各组(P<0.05),而其它3组之间差异无统计学意义。ROSC后PC组动物的存活率显著高于NT、SC和LC组(P<0.05或P<0.01)。结论:CPR后腹腔降温法可快速诱导亚低温和降低腹腔内温度,抑制肝脏内NF-κB的激活,降低血浆TNF-α和IL-6水平,减轻ROSC后全身炎症反应,延长动物存活时间。     

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.