在生理学研究中如何正确估计样本含量

目的：引起生理学研究人员对样本含量估计重要性的认识。方法：论述样本含量估计的意义及存在的问题,介绍常用的样本含量估计方法以及获取其他样本含量估计方法的途径。结果：清楚地表述了估计样本含量必需明白的基本概念、前提条件,并通过实例给出了两种场合下所需样本含量的估计过程和结果。结论：在估计样本含量时,必须明确资料将选用何种统计分析方法处理,并且应满足有关的前提条件,才能得到正确的估计结果。     

在生理学研究中如何正确把握试验设计三要素

目的：为从事生理学研究的科研工作者制订科学完善的试验设计方案提供一些有效的技术方法。方法：通过正面阐述三要素和正确把握每个要素的关键点;再通过分析已发表的学术论文中未妥善把握三要素的案例,为生理学研究工作者在制订课题设计方案时能正确把握三要素提供具有可操作性的方法。结果：呈现出试验设计三要素的具体内容,即"受试对象"、"影响因素"和"试验效应",并且,为如何正确把握每个要素提供具体措施。结论：试验设计的三要素是从事试验研究的"物质基础",正确把握它们是制订出科学完善试验设计方案的重要环节之一。     

脑水肿干湿重法的测定条件分析

目的：分析干湿重法测定脑水肿的温度条件和时间条件,探讨优化测量方案。方法：健康SD大鼠随机分为3组（n=4）：取脑组织称其湿重,分别连续烘烤16 h,用于不同温度条件下60℃、90℃、110℃的干湿重法脑水肿测定,检测不同时间点脑组织重量变化以及重量差值变化情况。结果：在不同温度下,随着时间的延长,脑组织重量逐渐减轻;温度在110℃条件下,9 h两次称量干重之差小于0.0002 g,已经烘烤至恒重;90℃温度条件下12 h至恒重;60℃温度条件下15.5 h至恒重。结论：干湿重法测量脑水肿的优化方案为110℃ 9 h、90℃ 12 h、60℃ 15.5 h,为脑水肿的测定提供实验依据。     

Flag和GFP双标记的BKCa通道α亚基表达质粒的构建、鉴定和序列分析

目的：采用重叠PCR法构建表达质粒,为下一步研究通道功能奠定基础。方法：在已有编码大电导钙激活钾通道（BK_Ca）通道α亚单位的表达质粒pcDNA3.1-hSlo的基础上,采用重叠PCR法构建Flag和GFP双标签标记的表达质粒pcDNA3.1-Flag-hSlo-GFP （Flag-hSlo-GFP）。结果：构建的表达质粒Flag标签插入BK_Ca通道的S1-S2胞外环,GFP标签连接BK_Ca通道的胞内C末端,测序结果证实质粒构建成功。结论：成功构建BK通道基因表达质粒Flag-hSlo-GFP,重叠PCR能够很好的用于长片段基因扩增和插入片段的实验。     

缺氧对稳定表达人淀粉样前体蛋白HEK293细胞的存活及阿尔茨海默病相关蛋白表达的影响

目的：探讨缺氧对稳定表达人淀粉样前体蛋白的HEK293细胞（HEK293-APP695）存活及相关蛋白表达的影响,为深入研究缺氧对阿尔茨海默病的调节作用提供稳定的细胞模型。方法：利用缺氧手套箱（0.3% O₂）处理HEK293-APP695细胞,CCK-8法检测细胞的存活情况;Western blot检测缺氧条件下阿尔茨海默病（AD）相关蛋白APP、APP-CTFs和BACE1的表达变化。结果：缺氧处理后,HEK293-APP695细胞的存活率明显下降,APP表达降低,其剪切体APP-CTFs表达升高。结论：缺氧导致APP剪切的增多,抑制细胞的存活,提示缺氧可能通过影响BACE1的活性在AD的发病进程中起重要的调节作用。     

老年人跌倒防控信息管理系统的建立及其应用

目的：探索建立老年人跌倒防控信息管理系统,并进行应用及效果评价。方法：依托物联网技术和信息化手段,将老年人跌倒综合防控策略从线下导入线上,建立集风险评估、远程教育、反馈评价于一体的老年人跌倒防控信息管理系统。运用该系统对126名干休所老年人（年龄≥ 60岁）进行评估,量化跌倒风险等级,对筛选出的84名高危老年人进行远程连续综合干预,干预措施包括分发宣传画册,制作宣教幻灯片及视频进行远程播放与讲解。采用自身前后对照,分析评价干预6个月后跌倒相关情况。结果：高危人群跌倒发生率由21.43%降至4.76%（P<0.01）;身体平衡能力和步态稳定性改善情况明显（P<0.01）;能够自觉进行正确防控行为的比例显著提高（P<0.01）;进行复杂行为时更有信心不会跌倒（P<0.01）;居家环境安全性整体提高（P<0.01）。结论：老年人跌倒防控信息管理系统可通过全面评估筛选出高危人群,实施远程连续综合干预,降低跌倒发生率,可进一步推广应用。     

瞬时感受器电位M2通道抗原位点的兔单抗特异性检测

目的：筛选特异性较高的抗体以推动对瞬时感受器电位M2（TRPM2）通道结构和功能的研究。方法：以野生型昆明种小鼠大脑皮层、人胚肾293细胞、未诱导的TRPM2细胞及四环素诱导的TRPM2细胞为标本,采用免疫印迹和免疫荧光方法,以被检测抗体为一抗,荧光分子结合的抗体为二抗,根据170kD（TRPM2通道蛋白分子量）位置上是否有特异性条带,检测兔单抗的特异性。结果：抗体98927对鼠源TRPM2通道有特异性,抗体40622,抗体98721,抗体98921对人源TRPM2通道有特异性,另外抗体98721对鼠源TRPM2通道的突变型有特异性。结论：作为分子探针,抗体98927、40622、98721、98921可用于TRPM2通道结构和功能研究。     

α-烯醇化酶基因干扰表达对籽鹅卵泡颗粒细胞增殖及凋亡的影响*

目的: 研究α-烯醇化酶(ENO1)基因干扰表达对籽鹅卵泡颗粒细胞增殖、凋亡和细胞周期的影响。方法: 原代培养籽鹅F1级卵泡颗粒细胞(复合培养),将ENO1基因干扰表达重组质粒转染至籽鹅卵泡颗粒细胞。实验分为四组:ENO1干扰表达组(RNAi)、无关序列干扰组(NC)、培养液组(Control)、转染试剂组(Lip)。流式细胞术检测干扰组和对照各组的凋亡率、细胞周期时相性。结果: ENO1基因干扰表达使籽鹅卵泡颗粒细胞增殖速度变慢,凋亡率增加,G2/M期颗粒细胞比例增高。结论: ENO1基因干扰表达可使籽鹅卵泡颗粒细胞周期发生G2/M期阻滞,诱导细胞发生凋亡,抑制细胞增殖。     

三七总皂苷对脊髓损伤大鼠运动功能恢复的作用

目的：探讨三七总皂苷对大鼠脊髓损伤（SCI）后运动功能恢复的作用。方法：正常SD大鼠随机分为5组（n=8）：正常对照组（Normal）、假手术组（Sham）、脊髓损伤（SCI）和脊髓损伤+三七总皂苷组（PNS）（n=8）。所有大鼠分别在造模前及造模后第1、3、7、14、21和28天接受运动功能评分（BBB）和运动诱发电位（MEP）检查,观察大鼠后肢运动功能的恢复情况。结果：造模后,Sham组、PNS组、SCI组BBB评分低于正常;MEP波幅低于正常;潜伏期较正常延长。PNS组与同期SCI组比较,第7、14、21、28天的BBB评分差异有统计学意义（P<0.05）;第7天、14天、21天、28天,MEP检查波幅（Amp）和潜伏期（Lat）组内有显著差异,并且与同期SCI组比较差异有统计学意义（P<0.05）。结论：三七总皂苷可促进大鼠SCI后运动功能的恢复。     

狼疮性肾炎患者肾组织中胎源性微嵌合体的表达及其临床意义

目的：探讨胎源性微嵌合体（FMc）与狼疮性肾炎（LN）的相关性。方法：收集生育过男胎的24例LN和24例肾小球轻微病变（GML）的女性患者肾活检组织,采用荧光原位杂交技术（FISH）检测细胞中的Y染色体,并收集患者孕育史、临床和实验室资料进行分析。结果：在9例LN和3例GML的肾组织中发现Y染色体阳性嵌合细胞,两组发生率的差异有统计学意义（P<0.05）,但在LN组,嵌合细胞的出现与蛋白尿、血清肌酐、抗双链DNA抗体、抗核抗体、补体C3、C4水平及SLEDAI评分等之间均无相关（P>0.05）。结论：FMc在女性LN患者肾组织中的发生频率高于对照组GML患者,但FMc的存在很可能与LN的发病、疾病活动性和进展性无关。     

在生理学研究中如何严格遵循试验设计四原则

目的：为从事生理学研究的科研工作者严格遵循试验设计四原则（以下简称四原则）提供一些有效的技术方法。方法：通过简单介绍"四原则的概念"、举例阐述"生理学研究中在落实四原则方面出现的常见错误和案例"和简单介绍"生理学研究中如何遵循四原则"来阐述遵循四原则的具体做法。结果：需要根据实际情况,选择合适的随机化方法,分层随机化是常用的好方法;应力求设置合理的对照组,才能真正体现出对照原则的价值;有根据地给出每个小组合适的样本含量,是试验结果具有重现性的基本保证;严格遵循随机、对照和重复原则,最终才能实现对比组之间达到极好的均衡效果,这说明均衡原则是一个把关的原则。结论：在生理学科研中,严格遵循四原则对保证生理学研究结果准确可靠至关重要。     

Educating the future generation of researchers: A cross-disciplinary survey of trends in analysis methods

Methods for data analysis in the biomedical, life, and social (BLS) sciences are developing at a rapid pace. At the same time, there is increasing concern that education in quantitative methods is failing to adequately prepare students for contemporary research. These trends have led to calls for educational reform to undergraduate and graduate quantitative research method curricula. We argue that such reform should be based on data-driven insights into within- and cross-disciplinary use of analytic methods. Our survey of peer-reviewed literature analyzed approximately 1.3 million openly available research articles to monitor the cross-disciplinary mentions of analytic methods in the past decade. We applied data-driven text mining analyses to the “Methods” and “Results” sections of a large subset of this corpus to identify trends in analytic method mentions shared across disciplines, as well as those unique to each discipline. We found that the t test, analysis of variance (ANOVA), linear regression, chi-squared test, and other classical statistical methods have been and remain the most mentioned analytic methods in biomedical, life science, and social science research articles. However, mentions of these methods have declined as a percentage of the published literature between 2009 and 2020. On the other hand, multivariate statistical and machine learning approaches, such as artificial neural networks (ANNs), have seen a significant increase in the total share of scientific publications. We also found unique groupings of analytic methods associated with each BLS science discipline, such as the use of structural equation modeling (SEM) in psychology, survival models in oncology, and manifold learning in ecology. We discuss the implications of these findings for education in statistics and research methods, as well as within- and cross-disciplinary collaboration.

A quantitative survey of >1 million published research articles reveals that while classical statistical methods remain in widespread use, multivariate statistical and machine-learning approaches have seen a significant increase; statistics curricula should be revised to take full advantage of these new analytical tools.     

A New Hematological Stain: I. Constituents and Methods of Use

The stain proposed by the author is quickly and simply prepared by mixing equal parts of the following permanent stock solutions:

The mixed stain is usable, for at least eight months, and is applicable to practically all hematological purposes: blood smears, fixed sections, frozen sections, and touch preparations. In the technics utilized to produce its action on preparations treated in different ways, the only variants are the methods for treating the cells, while the stain itself remains totally unchanged for all purposes. For blood smears fixed with methyl alcohol, the technic consists merely of pouring the stain on the slide, leaving it for 5 to 7 minutes, and then washing it off. On sections, a further process of differentiation with acid acetone is rapidly carried out. The various types of granules, including megakaryocyte and platelet granules, are clearly demonstrated. For frozen sections, the technic is extremely rapid, yet yields excellent differentiation.     

Twenty-Four-Hour Variations in the Sensitivity of Rat Aorta to Vasoactive Agents

The presence of time-dependent variations in the in vitro sensitivity of aorta preparations to either vasoconstricting or relaxing agents was investigated in rats maintained in light from 08: 00 to 20: 00 and in darkness from 20: 00 to 08: 00. Rat thoracic aorta rings were obtained from animals sacrificed at four different times of the day. The rat aorta was found to be more sensitive to the constricting effect of phenylephrine at 15: 00, and of 5-hydroxytryptamine at 21: 00. On the other hand, both endothelium-dependent and -independent relaxations were more remarkable at 03: 00 than at other times of the day. These variations represented significant circadian rhythms when analyzed by analysis of variance. Different in vitro responsiveness to these agents might reflect changes in the sensitivity and/or number of related receptors in vascular preparations. In conclusion, the circadian time of animal sacrifice to obtain vascular preparations constitutes an important aspect of the research method and a key determinant of findings. (Chronobiology International, 13(6), 465-475, 1996)     

Integration and scaling of UV-B radiation effects on plants: the relative sensitivity of growth forms and interspecies interactions

Aims The relative plant type sensitivity and selected community interactions under increased UV-B radiation where examined. Specifically, we investigated: (i) if there are differences among growth forms in regard to their sensitivity to UV-B radiation, (ii) if increased UV-B radiation influences the plant competitive balance in plant communities and (iii) the response mechanisms of the UV-B radiation-sensitive species that might increase their fitness.Methods To answer our research questions, we used a mechanistic model that, for the first time, integrated the effects of increased UV-B radiation from molecular level processes, whole plant growth and development, and community interactions.Important findings In the model simulations, species types exhibited different levels of sensitivity to increased UV-B radiation. Summer C₃ and C₄ annuals showed similar growth inhibition rates, while biennials and winter C₃ annuals were the most sensitive. Perennials exhibited inhibitions in growth only if increased UV-B radiation results in increases in metabolic rates. In communities, species sensitive to UV-B radiation may have a competitive disadvantage compared to resistant plant species. But, sensitive species may have a wide array of responses that can increase their fitness and reproductive success in the community, such as, increased secondary metabolites production, changes in timing of emergence and reproduction, and changes in seed size. While individual plants may exhibit significant inhibitions in growth and development, in communities, these inhibitions can be mitigated by small morphological and physiological adaptations. Infrequent or occasional increased UV-B radiation events should not have any lasting effect on the structure of the community, unless other environmental factors are perturbing the dynamic equilibrium.     

心肺运动试验用于冠心病诊断价值的临床研究*

目的: 评估心肺运动试验（CPET）相关指标对于冠心病的诊断价值。方法: 选择156例疑诊为冠心病的患者（病情相对稳定,年龄在18~80岁）行CPET、运动负荷心电图及冠状动脉造影,以冠状动脉造影结果为标准,分析CPET相关指标（峰值摄氧量％预计值Peak VO₂%pred、峰值氧脉搏％预计值Peak O₂ pulse%pred、摄氧量对应功率的做功效率（ΔVO₂/ΔWR）诊断冠心病的敏感度、特异度及其诊断价值。结果: Peak VO₂%pred的最佳截断点是≤69%pred,其诊断冠心病的敏感度为55.1%,特异度为77.0%,AUC为0.698。Peak O₂ pulse%pred最佳截断点诊断冠心病的敏感度为50.7%,特异度为72.4%,AUC为 0.58。ΔVO₂/ΔWR最佳截断点处其诊断冠心病的敏感度为44.9%,特异度为87.4%,AUC为0.647。Peak VO₂%pred及ΔVO₂/ΔWR敏感度远高于负荷心电图,差异有统计学意义（P<0.01）。结论: CPET部分指标诊断冠心病的敏感度优于运动负荷心电图,最佳截断点处特异度及诊断价值均较高,而且CPET本身也包括连续动态全程心电图分析,不建议各自分割使用,主张所有指标整体整合分析使用。CPET相关指标对冠心病诊断有预测价值,可以早期、较准确的诊断冠心病。     

A user's guide to animal welfare science

Here, I provide a guide for those new to the burgeoning field of animal welfare science as to what this comprehensive, relatively young discipline is all about. Drawing on all branches of biology, including behavioural ecology and neuroscience, the science of animal welfare asks three big questions: Are animals conscious? How can we assess good and bad welfare in animals? How can we use science to improve animal welfare in practice? I also provide guidelines for an evidence-based approach to welfare issues for policy makers and other users of animal welfare research.     

山西省健康成年人淋巴细胞亚群正常参考值范围

目的:建立山西省健康成人外周血淋巴细胞亚群的正常参考值范围,为机体免疫状态的分析和肿瘤患者的免疫评估提供理论依据。方法:选取山西省1 238例健康成人体检人群,采用流式细胞术测定外周血淋巴细胞亚群的绝对计数和相对计数。结果:确定了健康成人外周血淋巴细胞表达水平,并发现CD3~+T细胞相对计数和绝对计数、CD4~+T细胞相对计数、CD8~+T细胞相对计数和绝对计数、NK细胞相对计数和绝对计数、CD19细胞相对计数和绝对计数、CD4/CD8比值在不同年龄组间存在显著差异性(P0.05);不同性别之间CD8~+T细胞相对计数、CD4~+T细胞绝对计数和CD19细胞绝对计数无统计学意义,CD3~+T细胞、CD8~+T细胞、NK细胞相对计数和绝对计数、CD4~+T细胞、CD19细胞相对计数均存在显著差异(P0.05)。结论:初步建立了山西省健康成年人外周血淋巴细胞亚群参考值范围,为机体免疫功能的评价和肿瘤免疫治疗、诊断提供了参考依据。     

Marchi's Staining Method: Studies of Some of the Underlying Mechanisms Involved

Spinal cord of cat and rabbit was stained, after experimental lesions, by variations of Marchi's method. The following conclusions were drawn:

1. The presence of an oxidizing agent (K₂Cr₂O₇, NaIO₃, or KCIO₃) in the osmic acid solution is of primary importance and a preliminary oxidation in Mueller's fluid is unnecessary or even detrimental.

2. Acetic acid added to Marchi's fluid, accentuates the action of the oxidizing agent in restraining the staining of normal myelin.

3. Too high concentration of oxidizing agent or of acid may inhibit staining of degenerating myelin.

4. Marchi's and Busch's methods have been modified as follows: Fix one day in 10% formalin and transfer without washing to the staining mixture, either A or B. Staining mixture A: Marchi's fluid plus 1 to 3% glacial acetic acid. B: An aqueous solution containing KCIO₃ 0.25%, osmic acid 0.33%, and acetic acid 1%. Stain about one week. These methods worked on spinal cord and medulla, but cannot be recommended for brain.

5. The detrimental effects of long post mortem autolysis or of prolonged fixation in formalin may be counteracted to some degree by increasing the concentration of the acid in Marchi's fluid up to 5% or of the KCIO₃ up to 0.4% in the modified Busch's fluid.